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Molecular Targets of Epigallocatechin—Gallate (EGCG): A Special Focus on Signal Transduction and Cancer

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Abstract

Green tea is a beverage that is widely consumed worldwide and is believed to exert effects on different diseases, including cancer. The major components of green tea are catechins, a family of polyphenols. Among them, epigallocatechin-gallate (EGCG) is the most abundant and biologically active. EGCG is widely studied for its anti-cancer properties. However, the cellular and molecular mechanisms explaining its action have not been completely understood, yet. EGCG is effective in vivo at micromolar concentrations, suggesting that its action is mediated by interaction with specific targets that are involved in the regulation of crucial steps of cell proliferation, survival, and metastatic spread. Recently, several proteins have been identified as EGCG direct interactors. Among them, the trans-membrane receptor 67LR has been identified as a high affinity EGCG receptor. 67LR is a master regulator of many pathways affecting cell proliferation or apoptosis, also regulating cancer stem cells (CSCs) activity. EGCG was also found to be interacting directly with Pin1, TGFR-II, and metalloproteinases (MMPs) (mainly MMP2 and MMP9), which respectively regulate EGCG-dependent inhibition of NF-kB, epithelial-mesenchimal transaction (EMT) and cellular invasion. EGCG interacts with DNA methyltransferases (DNMTs) and histone deacetylases (HDACs), which modulates epigenetic changes. The bulk of this novel knowledge provides information about the mechanisms of action of EGCG and may explain its onco-suppressive function. The identification of crucial signalling pathways that are related to cancer onset and progression whose master regulators interacts with EGCG may disclose intriguing pharmacological targets, and eventually lead to novel combined treatments in which EGCG acts synergistically with known drugs.
Nutrients 2018, 10, 1936; doi:10.3390/nu10121936 www.mdpi.com/journal/nutrients
Review
Molecular Targets of Epigallocatechin—Gallate
(EGCG): A Special Focus on Signal Transduction and
Cancer
Aide Negri
1,†
, Valeria Naponelli
1,2,
*
,†
, Federica Rizzi
1,2,3
and Saverio Bettuzzi
1,2,3
1
Department of Medicine and Surgery, University of Parma, Via Gramsci 14, 43126 Parma, Italy;
aide.negri@unipr.it (A.N.); federica.rizzi@unipr.it (F.R.); saverio.bettuzzi@unipr.it (S.B.)
2
National Institute of Biostructure and Biosystems (INBB), Viale Medaglie d’Oro 305, 00136 Rome, Italy
3
Centre for Molecular and Translational Oncology (COMT), University of Parma,
Parco Area delle Scienze 11/a, 43124 Parma, Italy
* Correspondence: valeria.naponelli@unipr.it; Tel.: +39-0521-033790
These authors contributed equally to this work.
Received: 7 November 2018; Accepted: 4 December 2018; Published: 6 December 2018
Abstract: Green tea is a beverage that is widely consumed worldwide and is believed to exert
effects on different diseases, including cancer. The major components of green tea are catechins, a
family of polyphenols. Among them, epigallocatechin-gallate (EGCG) is the most abundant and
biologically active. EGCG is widely studied for its anti-cancer properties. However, the cellular and
molecular mechanisms explaining its action have not been completely understood, yet. EGCG is
effective in vivo at micromolar concentrations, suggesting that its action is mediated by interaction
with specific targets that are involved in the regulation of crucial steps of cell proliferation,
survival, and metastatic spread. Recently, several proteins have been identified as EGCG direct
interactors. Among them, the trans-membrane receptor 67LR has been identified as a high affinity
EGCG receptor. 67LR is a master regulator of many pathways affecting cell proliferation or
apoptosis, also regulating cancer stem cells (CSCs) activity. EGCG was also found to be interacting
directly with Pin1, TGFR-II, and metalloproteinases (MMPs) (mainly MMP2 and MMP9), which
respectively regulate EGCG-dependent inhibition of NF-kB, epithelial-mesenchimal transaction
(EMT) and cellular invasion. EGCG interacts with DNA methyltransferases (DNMTs) and histone
deacetylases (HDACs), which modulates epigenetic changes. The bulk of this novel knowledge
provides information about the mechanisms of action of EGCG and may explain its
onco-suppressive function. The identification of crucial signalling pathways that are related to
cancer onset and progression whose master regulators interacts with EGCG may disclose
intriguing pharmacological targets, and eventually lead to novel combined treatments in which
EGCG acts synergistically with known drugs.
Keywords: green tea catechins; epigallocatechin-gallate (EGCG); 67LR; cancer apoptosis; cell death;
chemoprevention; gene expression
1. Introduction
Green tea is produced from Camellia sinensis and it represents the second most consumed
beverage in the world after water, being used primarily in Asia and in the Middle East [1].
Several observational and intervention studies have demonstrated that green tea consumption
has beneficial effects on many human diseases, including obesity, metabolic syndrome,
neurodegenerative disorders inflammatory diseases, and cancer [2–6]. The major polyphenolic
component of dried green tea extracts is epigallocatechin-gallate (EGCG) EGCG is the most
Nutrients 2018, 10, 1936 2 of 24
abundant and biologically active catechin from green tea, accounting for at least 50% of the total
catechin content in green tea leaves [7]. The biological effects of green tea were initially ascribed to
pro- or anti-oxidative properties of catechins. Most of the studies have been conducted
administrating green tea extracts or pure EGCG. A typical weakness of many studies is related to
data collected in vitro and cell culture systems, following the administration of doses of green tea
extracts (or EGCG) much higher than those that were reached in human plasma after green tea
consumption. In vivo, administration of the equivalent of two or three cups of green tea leads to a
peak in the plasma levels of tea catechins in the sub-micromolar range in humans [8,9].
Several in vitro, in vivo, and clinical studies have shown multiple EGCG anticancer actions.
Among them there are anti-proliferative, pro-apoptotic, anti-angiogenic, and anti-invasive functions
[10]. Furthermore, EGCG has been observed to impair other processes that are involved in
carcinogenesis as inflammation, oxidative stress and hypoxia and to target tumor microenviroment
components (e.g., cancer stem cells, fibroblasts, macrophages, and microvasculature) [11]. In several
in vitro and in vivo cancer types, EGCG has been shown to act synergistically with other natural
compounds (e.g., curcumin, ascorbic acid, quercetin, genestein, caffeine) [10,12] and it has also been
testing in combination with currently used chemotherapeutic drugs (e.g., doxorubicin, cisplatin,
sunitinib) [13–16]. Furthermore, in order to improve EGCG bioavailability and stability, novel
formulations of the catechin encapsulated in nanoparticles have been developed [17–20].
Even if the anti-tumoural effect of green tea catechins (and specifically EGCG) has been
extensively demonstrated in vitro, their molecular and cellular mechanisms are not yet completely
understood [21,22].
The anti-cancer effect of EGCG and green tea extracts is mediated through several mechanisms,
including stimulation of anti-oxidant activity and activation of detoxification system [23,24],
alteration of the cell cycle [25], suppression of mitogen-activated protein kinase (MAPK) and
receptor protein kinase (RTKs) pathways [26,27], inhibition of clonal expansion of the
tumour-initiating stem cell population [28], and production of epigenetic changes in gene expression
[29]. These mechanisms (reviewed recently in [30–32]) are not completely understood yet. Green tea
catechins are thought to function both as powerful radical scavengers, in particular, under increased
oxidative stress conditions [33], and as ROS generators leading to the inhibition of cancer cell growth
through the induction of apoptotis [24,34]. Moreover, they have been shown to induce apoptosis in
several ways, such as modulating pro- and anti-apoptotic protein (Bax, Bcl-2, Bcl-XL) and cell cycle
regulator proteins (cyclins, CDKs) [35]. Green tea catechins are also able to target genes and proteins
that are associated with cell proliferation and apoptosis, including RTKs (receptor tyrosine kinases).
Several studies described the inhibitory effect of green tea catechins on these receptors and on
Ras/extracellular signal-regulated kinase (ERK)/MAPK and phosphatidylinositol 3-kinase
(PI3K)/Akt, which are RTKs-related downstream pathways that are often constitutively activated in
tumor cells. EGCG negatively modulates the expression of various transcription factors, including
Sp1, AP-1, and NF-kB preventing cancer formation [36,37]. Another mechanism that can explain the
pleiotropic effects exerted by green tea catechins is represented by the epigenetic changes in gene
expression and chromatin organization. The major epigenetic mechanisms are DNA methylation,
histone modifications, and expression of noncoding regulatory micro RNA (miRNAs). Green tea
catechins can induce an epigenetic reactivation of genes silenced during carcinogenesis or an
epigenetic downregulation of oncogenes through the inhibition of DNA methyltransferases
(DNMTs) or histone deacetylases (HDACs) activity and the reduction of their expression [38,39].
Micromolar concentrations of EGCG have been shown to exert a wide array of different effects
in a cancer cell. The current understanding is that catechins may either interact with a single critical
regulator affecting the activity of key enzymes that are involved in important pathways, or by
hitting multiple targets in parallel, thereby modulating different pathways simultaneously [40,41].
First it is necessary to identify proteins that bind to catechins with high affinity, which may
represent the master regulators controlling one or multiple pathways. Using in vitro models, several
research teams have identified proteins that are targeted by EGCG. Among these are vimentin, Fyn,
ZAP70, insulin-like growth factor 1 receptor, and glucose regulated protein 78 kDa [42–46].
Nutrients 2018, 10, 1936 3 of 24
However, the functional effect of green tea catechins on the target protein activity has been
demonstrated only at much higher concentrations of EGCG than Kd values, probably because of the
non-specific binding of EGCG to other proteins competing for the target [47]. In Table 1 are listed the
principal EGCG molecular targets that were identified in cancer cells.
We think that EGCG-protein binding can be important for the beneficial effect of green tea
catechins. Green tea catechins bind to a plethora of proteins and the process of the interaction is
highly dependent on the folding status and on the conformational properties of the target protein. In
this review, we decided to take into consideration only few proteins that, after direct binding to
EGCG, alter and affect their downstream pathways promoting anti-cancer effects. These data could
be used for a rational drug design of green tea catechins derivatives exploitable for more specific and
effective anti-cancer therapies. We will focus particularly on the onset and progression of cancer,
describing and discussing the possible molecular mechanisms through which catechins exert their
action.
Table 1. Epigallocatechin-gallate (EGCG) molecular targets that are involved in cancer onset and
progression.
Cell Cycle,
Proliferation
& Survival
Apoptosis &
Cell Death
Motility,
Invasion and
Metastatization
Inflammation Epigenetic
Control Others
p16 [48]
Bax [49]
MMP-2 * [50]
FcεRI [51]
DNMT1 * [39]
DAPK1 [52]
p18 [35] Bad [53] MMP-9 * [50] IL-8 [54] DNMT3A
[48] MRLC [55]
p21 [48] Bak [56] MMP-14 [57] IGF-1R * [45]
DNMT3B *
[39] MYPT1 [55]
p27 [56] Bcl-2 * [58] uPA [59] VEGF [60] HDAC1 * [39] eEF1a [61]
Cyclin D [56]
Bcl-xl [53]
PAI
1 [59]
CSF-1 [62]
HDAC2 [63]
ID1 [64]
Cyclin E [35] Bcl-xs [56] E-cadherine
[39] CCL-2 [62] HAT [65] RAR-β [39]
Cyclin A [66]
Caspase3 [56]
SLUG [67]
COX-2 [60]
hTERT [68]
HSP70 [53]
Cyclin B [66]
Caspase8 [69]
SNAIL1 [70]
iNOS [71]
EZH2 [72]
HSP90 * [73]
CDK4 [56] Caspase9 [56] Vimentin * [42] eNOS [74–78] GRP78 * [46]
CDK6 [56] Apaf-1 [53] Twist [79] PECAM-1
[80]
CDK2 [35] Puma [56] N-cadherine
[79] miR-16 [62]
CDK1 [66] XIAP [53] HIF-1α [60]
let-7b miRNA
[81]
Erk1/2 [56] Cytochrome
C [56] β-catenin [54] miR-210 [82]
Pin * [83] p53 [84] Wnt [54] miR34a [85]
PPA2 [86] Survivin [87] TIMP-3 [72] miR145 [85]
PKA [86]
Fas [69]
miR200c [85]
STAT [12]
DR5 [69]
ZAP70 * [44]
AR [65] PARP [88] TRAF-6 * [89]
67LR * [90] Oct4 [85]
FcεRI [51]
Sox2 [91]
EGFR [92] Notch1 [85]
HGFR [93] Nanog [85]
TGFR-II * [94]
CD133 [95]
cGMP [74] [96]
cAMP [86]
P-glycoprotein
Nutrients 2018, 10, 1936 4 of 24
[88]
NF-kB [97]
c-Myc [98]
FOXO3a [99]
GSK-3β [98]
PI3K [100]
AKT [100]
PKC-δ [74]
JAK-1/2 [12]
Src [57]
CK1α [98]
p38 MAPK
[56]
JNK [56]
* EGCG direct interactors.
2. 67-kDa Laminin Receptor Signalling Pathways
One of the most interesting targets of EGCG action is the 67-kDa laminin receptor (67LR), a
non-integrin cell surface receptor whose expression has been shown to be increased in several
cancers, such as blood, prostate, breast, gastric, and colon [101–106]. The receptor expression is
usually correlated with drug resistance, and it contributes positively to cancer cells viability, tumour
progression, metastatic diffusion, and neo-angiogenesis [101,102,107,108].
In 2004, Tachibana et al. identified for the first time the 67LR as a specific EGCG membrane
receptor using surface plasmon resonance. The study revealed that 67LR was able to bind EGCG
with a Kd value of 39.9 nM. This interaction enabled EGCG to reduce the growth of the lung cancer
cell line A549 [90], thus exerting anticancer activity. Other green tea components, such as caffeine,
quercetin, epicatechin (EC), and epigallocatechin (EGC) were tested for binding to 67LR, but none
were specific ligands of the receptor or showed tumour suppressive effects [90]. Therefore, EGCG
appears to be the only catechin able to bind 67LR. Subsequently, a putative EGCG binding site
corresponding to the region between the residues 161 and 170 of the receptor has been identified
[109]. The direct binding between EGCG and 67LR has been confirmed in prostate cancer cells by Yu
et al. [104]. Using MVD (Molegro Virtual Docker, an integrated platform for predicting protein
ligand interactions), these authors identified a binding site for EGCG with the same sequence of the
laminin tyrosine-isoleucine-glycine-serine-arginine (YIGSR) peptide, corresponding to the 929–933
sequence of β1 chain of 67LR [104].
Many studies have explored the signalling cascades that are triggered by EGCG-67LR
interaction, some of which will be discussed in this review. In many cases, the tumour suppression
pathway affected ordered microdomains of the cell membrane known as lipid rafts, where the 67LR
has been located [110]. Lipid rafts differ from the surrounding membrane, because their composition
is enriched in specific lipids (sphingomyelins and glycosphingolipids) and cholesterol, which are
tightly packed to form liquid ordered assemblies [111]. Lipid rafts are dynamic, heterogeneous
structures whose composition is extremely variable, not only in relation to the lipid and sterols
content, but also because of the several proteins that can be recruited (e.g., BCR, FcεRI) or harboured
(e.g., Scr tyrosin kinases) [112–115]. Lipid rafts are rich in tyrosine kinase receptors (RTKs), such as
EGFR [116–118], IGF1R [119], and HER2 [120]. These receptors have been found to be inhibited by
EGCG in several in vitro and in vivo cancer models (e.g., colon, lung, liver and breast cancers)
[92,121–123]. The functional proteins recruited by lipid rafts allow these structures to play complex
roles. Lipid rafts can float within the plasma membrane or can cluster in larger and stabilized
platforms in response to different stimuli. In most cases, the lipid rafts clustering allows for the
activation of the proteins [124]. Furthermore, modifications in lipid rafts/protein interaction can lead
to alterations in lipid and sterol content, which can, in turn, influence lipid raft functions. Thanks to
their capability to interact with several cellular and molecular factors as caveolae [111], viruses [125],
Nutrients 2018, 10, 1936 5 of 24
bacteria, inflammatory molecules [126,127], and growth factors [128–130], these microdomains are
involved in a plethora of biological functions, like cell polarization, membrane trafficking [111],
pathogen internalization [126], and regulation of a wide spectrum of signal transduction pathways
[131]. Because most of these pathways can control cancer development, progression, rate of cell
proliferation [114], migration, invasion [132,133], and apoptosis [134], lipid rafts composition and
functions have received much attention. In addition, many anti-cancer agents (e.g., edelfosine, avicin
D, resveratrol) exert their anti-tumour activity, at least in part, by altering or disrupting the structure
of lipid rafts [135,136].
EGCG has been found to bind to the plasma membrane by interacting with the lipid rafts. The
first evidence of this association was shown in the basophilic cell line KU812, where the suppressive
action of EGCG on the expression of the high-affinity immunoglobulin E receptor (FcεRI) was
triggered by direct binding to lipid rafts. This was mediated by the inhibition of Erk1/2 kinases
phosphorylation and activation [51]. Shortly after, the same research team observed that the
down-regulation of FcεRI was driven by EGCG through binding to 67LR, a receptor associated with
lipid rafts [110]. Others reported that the EGCG inhibitory effect on EGFR in colon cancer cell line
HT29 [92], and on HGFR in prostate cancer cell line DU145 [93] was mediated by the alteration of
lipid rafts. The collection of signalling pathways affected by lipid rafts structure/function via
EGCG/67LR quickly increased in number, as reported in several tumour models, such as multiple
myeloma, mammary and epidermiod carcinoma, and chronic myeloid leukemia [52,137–139].
2.1. Lipid Rafts-Mediated Apoptosis
2.1.1. EGCG/67LR/Akt/eNOS/NO/cGMP/PKCδ/aSMase Pathway
Together with the inhibition of cell proliferation, migration, and angiogenesis, induction of
apoptosis is one of the main mechanisms through which EGCG exerts its anti-tumour activity
[140,141]. Several studies reported that EGCG is able to affect the expression and function of
anti-apoptotic factors (e.g., Bcl-2, Bcl-xl) and to up-regulate pro-apoptotic molecules (e.g., Bax,
caspase-3) in several cancer models [58,142–144]. However, the mechanisms through which EGCG
modulates key cell death regulators are not completely understood. Some studies reported that 67LR
plays a relevant role in triggering apoptosis after binding its ligand, EGCG, in haematological
malignancies, such as acute myeloid leukemia and multiple myeloma [145,146]. More recently, a
signalling pathway inducing EGCG/67LR-dependent apoptosis through the activation of protein
kinase Cδ (PKCδ), acid sphingomyelinase (aSMase), and lipid rafts clustering has been described in
multiple myeloma models [137] (Figure 1). The enzyme aSMase is responsible for the catabolism of
sphingomyelin (SM) and is known to be part of the signalling cascades that mediates lipid
raft-dependent apoptosis [147]. It can be activated in response to external pro-apoptotic stimuli as
physical agents (e.g., radiation, UVA light) [148,149], anti-cancer drugs (e.g., cisplantin, doxorubicin)
[150,151], and pro-apoptotic receptors (e.g., Fas, TNF-R) [147,152]. One of the best described
mechanisms of aSMase activation is triggered by Fas receptor. The binding between death receptor
Fas, harboured in lipid rafts [153], and its ligand FasL lead to the recruitment of adaptor
Fas-associated protein with death domain FADD, which in turn recruits and activates pro-caspase 8.
The final death-inducing signalling complex (DISC) then activates aSMase, which migrates from the
cytoplasmic compartment to lipid rafts, where it generates the sphingolipid ceramide from SM [154–
156]. In response to ceramide generation, cholesterol is displaced from lipid rafts, thus leading to an
increase of membrane fluidity [157]. Ceramide plays a role as second messenger in the signal
transduction, inducing lipid raft clustering and the stabilization of DISC complex, amplification of
Fas/FasL signalling, finally leading to apoptosis [155,158,159]. A similar mechanism has been
hypothesized in the case of cervical, prostate and colon cancer, where EGCG administration induces
cell apoptosis through aSMase activation and ceramide increase [160–162].
Studies on multiple myeloma cell lines in vitro and in vivo, in patients or murine models,
showed that the activation of 67LR through EGCG binding induces the activation of PKCδ after
phosphorylation of Ser664. Activation of PKCδ leads, in turn, to aSMase activation, and finally to cell
Nutrients 2018, 10, 1936 6 of 24
apoptosis [137]. These authors pointed out that treatment with 5 µM EGCG led to an increase of
nitric oxide (NO) [74]. NO is an inorganic signalling messenger triggering a wide range of cellular
pathways. The increase in NO levels is due to the activation of endothelial nitric oxide synthase
(eNOS), after phosphorylation in the residue Ser1177 by Akt kinase [74]. Production of NO causes an
increase of cGMP, produced by NO-dependent soluble guanylate cyclase (sGC) activation, and then
the phosphorylation of PKCδ [74] (Figure 1). Furthermore, more recently, it has been observed that
the anticancer agent coptisine induces apoptosis in hepatocellular carcinoma (HCC) cells via the
67LR/cGMP pathway [163]. Conversely, several studies reported that EGCG negatively regulates
eNOS/NO production in different cancer types [75–78] and also sGC/cGMP amount [96].
However, the fact that administration of 5 µM EGCG was sufficient to enhance NO production,
but not a significant increase of cGMP levels to induce cell apoptosis, gives rise to the question of
whether other factors might interfere with cGMP-mediated aSMase activation (Figure 1). Enzyme
phosphodiesterase 5 (PDE5), one of the major cGMP negative regulators, was found to be highly
expressed in multiple myeloma patients as compared to healthy donors, suggesting that PDE5 could
be a target for a possible combinatorial therapy with 5 µM EGCG [74]. This experimental approach
has been implemented. The combined treatment of PDE5 inhibitor Vardenafil and 5 µM EGCG
caused a strong reduction of cell viability not only in multiple myeloma, but also in other models as
prostate, gastric, pancreatic, breast cancer, and in acute myeloid and chronic lymphocytic leukemia
cell lines [74,164,165]. Vardenafil and EGCG synergistic action has been found to cause a significant
reduction of IC
50
of EGCG [74,164,165]. The tumour suppressive effects of the combinatorial therapy
have also been confirmed in vivo in xenograft murine models of multiple myeloma, treatment that
resulted in the reduction of tumour volume and increased survival without hepatotoxicity, a
possible side effect of high EGCG administration [74]. In this model, controls (namely cell lines and
primary cultures, as well as healthy animal models), were not affected by EGCG alone or in
combination with Vardenafil.
Figure 1. EGCG modulates cell division and apoptosis via 67LR. EGCG binding to 67-kDa laminin
receptor (67LR) activates apoptosis program through enhanced nitric oxide (NO) and cGMP
production, acid sphingomyelinase (αSMase) activation and ceramide generation. Ceramide
metabolization in sphingosine-1-phosphate (S1P) reduces cell apoptosis. EGCG binding to 67LR
inhibits via eukaryotic translation elongation factor 1a (eEF1A) cell cytokinesis inducing myosin
phosphatase target subunit (MYPT1) dephosphorylation and activation and myosin II regulatory
light chain (MRLC) dephosphorylation and inactivation.
Nutrients 2018, 10, 1936 7 of 24
2.1.2. EGCG/67LR/Ceramide/SphK1/S1P Pathway
The formation of larger platforms of cholesterol-enriched lipid rafts in cancer cells is often
associated with aberrant activation of RTKs, resulting in increased proliferation, survival, and
metastatic spread [166]. Instead, ceramide causes cholesterol displacement from lipid rafts,
formation of ceramide-enriched lipid rafts, and induction of cell apoptosis [147]. Therefore, ceramide
catabolism/degradation may produce anti-apoptotic effects. Ceramide can be deacetylazed and
converted to sphingosine, which can be phosphorylated to sphingosine-1-phosphate (S1P) by the
sphingosine kinase 1 (SphK1), an enzyme that is highly expressed in several cancers. The S1P can
activate protein G-coupled receptors that can in turn activate pro-survival and anti-apoptotic
signalling. In prostate cancer models, treatment with high doses of EGCG (75 µM) suppressed
tumour growth in vitro and in vivo through the inhibition of SphK1/S1P signalling [167] (Figure 1).
The lesson from these data is that, in a particular cell system, a correct balance between
ceramide synthesis and catabolism is fundamental [168,169]. Treatment with 1 µM and 5 µM EGCG
in the multiple myeloma cell line U266 caused the induction of aSMase activity [52]. However,
ceramide accumulation has been observed only after giving high concentrations of EGCG (10 µM
and 20 µM EGCG) [52]. Treatment with these high doses of EGCG leads to the disruption of
cholesterol-enriched lipid rafts and the inhibition of phosphorylation and the activation of several
RTKs (e.g., EGFR, ErbB2, ErbB3, HGFR, IGF1R, Mer, and Flt3). IGFR inhibition has been
demonstrated to be dependent on 67LR and aSMase expression [52]. Because the amount of SphK1
has been found to be increased in multiple myeloma cell lines and specimens from patients, the
combinatorial treatment of 5 µM EGCG and SphK1 inhibitor Safingol was tested. The data
demonstrate that the combination of the two drugs caused an increase in ceramide content, the
disruption of cholesterol-enriched lipid rafts, inhibition of RTKs phosphorylation, and finally an
increase of cell apoptosis [52]. Furthermore, the combination treatment also affected another cell
death mediator that was activated by ceramide, the death-associated protein kinase 1 (DAPK1),
causing the de-phosphorylation of DAPK1 inhibitory residue Ser308 and leading to its activation
[52].
Thus, like the inhibition of PDE5 with Vardenafil, the simultaneous action on two related
pathways employing two agents in combination, produced a synergistic effect that strongly reduced
the IC
50
of EGCG [52]. The onco-suppressive action of the double treatment (EGCG plus Safingol)
has been found effective in vitro in acute myeloid leukemia, chronic myeloid leukemia, and in
chronic lymphocytic leukemia models [170]. Absence of toxicity of the combined therapy has also
been shown in vivo [52].
2.2. Cancer Cell Growth Inhibition
2.2.1. EGCG/67LR/eEF1a/MYPT1/MRLC Pathway
EGCG can exert anticancer functions inducing cell cycle arrest. Several studies reported the
blockade of the cell division cycle by EGCG administration in G0, G1, S, and G2 phases. EGCG may
act through the indirect downregulation of pro-proliferative factors, such as cyclin D1, cyclin E,
cyclin A, cyclin B, CDK4, CDK6, CDK2, and CDK1, as well as by the upregulation of
anti-proliferative effectors, such as CDK inhibitors p27, p21, p16, and p18 [35,48,56,66,171,172]. In
addition, EGCG has been found to act on cytokinesis, a critic step of cell division, by interacting with
67LR receptor [55,61,173].
Cytokinesis is the final step of cell division, leading a mother cell to be divided into two
daughter cells. Early events of the process require the formation of an actomyosin ring, also known
as contractile ring, that allows the formation of the cleavage furrow at the equator of mitotic cells
[174,175]. Generation of the furrow enables the equal division of genetic material between the two
forming cells and their subsequent separation. The interaction between actin filaments (F-actin) and
myosin motors is controlled by different processes among which is the
phosphorylation/dephosphorylation of the myosin II regulatory light chain (MRLC). Myosin II is
one of the main motors involved in cytokinesis, activated through MRLC phosphorylation at
Nutrients 2018, 10, 1936 8 of 24
Ser19/Thr18 by kinases, such as MLCK, ROCK, and Citron kinase [176]. Ser19 phosphorylation
favours the interaction with F-actin, the contractile ring formation, and filaments assembly. A
di-phosphorylation seems to be involved in the assembly of filaments, but the role of
phosphorylation in Thr18 alone is less clear [177–179]. Conversely, MRLC dephosphorylation in
Ser19 or Ser19/Thr18 by the myosin phosphatase leads to myosin inactivation. The MRLC activity is
also indirectly regulated through the phosphorylation of myosin phosphatase itself. When the
largest region of myosin phosphatase, called myosin phosphatase target subunit (MYPT1), is
phosphorylated in at least one of the inhibitory sites (e.g., Thr696, Thr853), its activity is inhibited,
and, as a consequence, MRLC remains active, thus providing a positive signal triggering cytokinesis
[180].
EGCG has been found to be able to interfere with the cytokinesis of HeLa cells through its
action on MRLC phosphorylation status, thereby affecting the cellular growth [55]. EGCG activates
the signalling cascade that is responsible for the impaired MRLC phosphorylation through binding
to its membrane receptor 67LR [55] (Figure 1). At first, it was reported that the treatment of HeLa
cells with 10, 20, and 50 µM EGCG resulted in the disruption of stress fibers, reduction of the
contractile ring formation, increment of cells blocked in G2/M phases, and inhibition of cell growth
[55]. Further analyses revealed that EGCG treatment also caused, via 67LR, a reduction in single
Ser19 and in double Ser19/Thr18 MRLC phosphorylation, which effects on MRLC phosphorylation
might reasonably trigger the effects shown on cell division and growth [55]. Under similar
conditions, EGCG was also found to decrease the phosphorylation of MYTP1 at inhibitory site
Thr696 both in vitro and in vivo, thus preventing myosin phosphatase inactivation. According to the
literature, the ability of EGCG to interfere with MRLC phosphorylation could be the indirect
consequence of MYPT1 loss of inhibition [55]. Recently, another factor has been added to the
members of the EGCG signalling pathway, believed to be responsible for impaired cancer cell
cytokinesis: the eukaryotic translation elongation factor 1a (eEF1a), which has been found to be
necessary to enable EGCG to alter MYPT1 phosphorylation status [61]. eEF1a is mainly known as a
component of the eukaryotic translation machinery, but it also takes part in other cellular processes,
such as senescence, oncogenic transformation, and cell proliferation [181–183]. eEF1a is able to bind
to MYPT1 and F-actin [184]. In vitro and in vivo experiments demonstrated that no significant
reduction in MYPT1 and MRLC phosphorylation, actin disassembly and cell proliferation was
observed after EGCG administration in eEF1a knockout models [61]. This evidence has been further
corroborated by the observation that when eEF1a levels are restored and 67LR is absent, the effects
that are described above disappear as well. Thereby, eEF1a is thought to be downstream of 67LR and
upstream of MYPT1 in the signalling pathway that is triggered by EGCG [61] (Figure 1).
2.2.2. EGCG/67LR/cAMP/PKA/PP2A Pathway
67LR surface receptor is involved in the selective anti-tumour activity exerted by EGCG in
melanomas. Tsukamoto et al. identified protein phosphatase 2A (PP2A) as a downstream target of
67LR in melanoma cells [86]. PP2A is a Ser/Thr phosphatase that is involved in important cellular
processes, such as proliferation, signal transduction, and apoptosis, and it is considered to be a
tumor suppressor that is functionally inactivated in cancer [185,186].
By performing functional genetic screening, Tsukamoto and colleagues showed that EGCG
binding to 67LR receptor induces PP2A activation mediated by the cAMP/PKA pathway [86], which
led to the suppression of melanoma tumor cell growth. Even though the direct interaction between
EGCG and PP2A was demonstrated using very high EGCG concentrations [84,187,188], 1 µM EGCG
was sufficient to activate 67LR/PP2A pathway. PP2A directly interacts with p70S6k and
down-regulates mTOR signaling [189], which is usually aberrantly activated in melanomas.
Therefore, it represents an important contribution to chemotherapeutic resistance of commonly used
BRAF inhibitor treatment. The EGCG-activating 67LR/PP2A pathway exerts a strong synergistic
effect with PLX4720, a BRAF inhibitor, in drug-resistant melanomas.
Another effect that is mediated by the 67LR/PP2A signaling is the activation of Merlin, a tumor
suppressor protein that is encoded by the NF2 gene at physiological concentrations of EGCG, as low
Nutrients 2018, 10, 1936 9 of 24
as 1 µM [86]. Merlin activity seems to target cell surface RTKs and adhesion/extracellular matrix
receptors, regulating cell proliferation, survival and motility [190]. PKA, p21-activated kinase 1 and
2 (PAK 1/2), or MYPT can activate Merlin by dephosphorylation at Ser-518. In the study by
Tsukamoto et al. [86], EGCG was demonstrated to be an activator of Merlin via 67LR/PP2A pathway.
In prostate cancer cell lines the absence or inactivation of Merlin contributes to tumor development
and progression toward a highly invasive and chemo-resistant state [191–193].
Recently published data show that 10 µM EGCG up-regulates let-7b miRNA expression not
only in melanoma cell lines, but also in metastatic melanoma tumours in vivo [81]. miRNAs are
non-coding RNAs transcripts that are able to regulate fundamental biological activities related to
mRNA degradation or translational inhibition [194]. Yamada et al. demonstrated that 67LR is
involved in the EGCG-elicited let-7b increase, which leads to the inhibition of melanoma tumor
progression [81]. Let-7b recognizes multiple target genes that are related to tumor progression, such
as the high mobility group A2 (HMGA2), decreased in EGCG-treated melanoma cells [81], or Ras
[195,196].
Furthermore, the data indicated that PP2A inactivation caused the induction of let-7b, which is
generally down-regulated in cancer (including melanoma and prostate cancer) [81,197], even if it is
not clear whether let-7b transcription or let-7b processing is modulated by EGCG-induced PP2A
activation.
Zhou et al. confirmed that EGCG induced miRNAs profile changes in a mouse model of lung
tumor. They highlighted that the miRNAs affected by EGCG and target genes are different from
those that were previously identified by in vivo studies [198].
2.3. Modulation of Cancer Stem Cells Properties
EGCG was shown to affect the survival of cancer stem cells (CSCs). EGCG inhibits CSCs growth
and stemness in several malignancies, such as breast [199], lung [54,200], colorectal cancer [85],
osteosarcoma [14], and neuroblastoma [201].
Kumazoe M. et al. [202] describe the effects of EGCG on the features of pancreatic CSCs (i.e., the
capability to form colonies and spheroids) through the activation of the EGCG/67LR/cGMP axis. The
same research team had observed that spheroid formation in pancreatic CSCs colonies was inhibited
by cGMP targeting of the Forkhead box O3 (FOXO3)/CD44 axes [203]. Transcriptional factor FOXO3
is known to be a cancer suppressor, but it also induces the high expression of CD44, a master
regulator (and also a marker) of CSCs [202]. FOXO3 has been shown to be a direct target of EGCG in
tumours, like pancreatic and breast cancer. In pancreatic cancer treatment with EGCG suppressed
tumour growth, accompanied by FOXO3 downregulation [99]. By contrast, in breast cancer, a
positive regulation of FOXO3 exerted by the EGCG has been described [70,204,205]. Although the
reported modulations seem to be opposite, the action of EGCG on FOXO3 seems to lead to cancer
suppression altogether. Recently, the role of EGCG in inhibiting cancer stem cells (CSC) growth and
altering their features is emerging [54,95,199]. According to this literature, EGCG seems to act by
downregulating CD44 expression in tumours, like non-small cell lung cancer and pancreatic cancer
[200,202]. In pancreatic cancer cell lines expressing CD44, the isoform 3A of the enzyme
phosphodiesterase (PDE3A) is highly expressed [202]. Like other members of the same family,
PDE3A is a negative regulator of cGMP [206]. In pancreatic cancer cells, low EGCG administration
did not lead to a significant increase in cGMP amount, or to the reduction of colony and spheroid
formation [202]. Further experiments were conducted using low doses of EGCG combined with the
administration of a PDE3A inhibitor, Trequinsin. The combination therapy decreased the protein
levels of FOXO3 and CD44, caused an increase of cGMP, and a strong reduction in the CSCs
capability to form both colonies and spheroids. The combination of EGCG and Trequinsin is
synergistic and it reduces the IC
50
of EGCG, thus allowing for its use at physiological concentration.
These observations have also been confirmed in vivo [202]. Surprisingly, as for the other signalling
pathway that is discussed above, the effects of EGCG alone, or in combination with other agents, are
always specific for cancer cells, and they do not affect normal cells. This highly specific effect of
EGCG is still waiting for an explanation.
Nutrients 2018, 10, 1936 10 of 24
3. Other EGCG-Interacting Proteins
Another interesting protein that was shown to interact directly with green tea catechins is the
human peptidyl prolyl cis/trans isomerase (Pin1). Pin is a protein with two domains: an N-terminal
WW-domain and a C-terminal PPIase domain; both are necessary for its function. Although many
PPIases have been identified some with an established role in cancer, only Pin1 acts distinctively and
specifically on phosphorylated proteins. Pin1 catalyzes the cis/trans isomerization of the peptidyl
proline bond of proteins. This activity causes major changes in the conformation of the target
protein, with a consequent alteration of its function or stability. In this way, Pin1 affects and
modulatse different pathways involving kinase-dependent signaling, such as NF-kB,
activator-protein 1 (AP-1), nuclear factor of activated T cells (NFAT), or b-catenin [207]. Pin1 has
been demonstrated to have a major role in oncogenic signaling [208,209] and is highly expressed in
several cancers [210,211], including prostate cancer [212].
Urusova et al. used crystallographic and biochemical data to show that EGCG interacts directly
with both the PPase and WW domains of Pin1, which inhibits its tumour-promoting activity.
Therefore, Pin1 represent a possible target for anti-cancer therapies [83,213]. The dissociation
constant of EGCG and Pin1 has been calculated as 21 µM, both by protease-coupled and isothermal
titration calorimetric assays: this value is similar to the concentration of EGCG that was found to
exert anti-cancer effects in experimental cancer models [40]. Since the Kd value that resulted was
quite high, the interaction between EGCG and Pin1 was described as “not strong”. Urusova and
colleagues crystallized the Pin1-EGCG complex, resolving its structure at 1.9 Å resolution by X-ray
diffraction. The crystal structure has revealed that a molecule of EGCG was bound to Pin1 WW
domain (aminoacids 1–31), which is responsible for the interaction with the substrate, while another
molecule of EGCG was bound to the Pin1 PPIase domain, necessary for the isomerization reaction. A
recent study demonstrated that galloyl group in EGCG is required for Pin1 inhibition [214]. Binding
between EGCG and Pin1 in solution has been studied recently by combining fluorescence spectrum,
far-UV circular dichroism spectrum with molecular dynamics simulations. The analysis of the
binding energy confirmed the strong inhibitory effect that is exerted by EGCG on Pin1 activity [215].
To analyze the functional consequence of Pin1-EGCG binding, Urusova and colleagues used
mouse embryonal fibroblasts (MEF) collected from PIN1 KO and WT mice, and showed that Pin1
expression is required for EGCG (10–40 µM) inhibitory effect on MEFs growth. Furthermore, the
formation of the EGCG-Pin1 complex prevented the binding of the Pin1 substrate c-Jun. Finally,
EGCG effect on transcriptional regulation of AP-1 and NF-kB has been shown to be mediated by
Pin1 [83].
Green tea catechins are mainly believed to prevent cancer. However, several epidemiological
studies suggest that their activity also works against cancer progression; the interaction of EGCG
with proteins that are involved in cancer progression and metastatic spread has been considered.
One of the effects exerted by EGCG is the inhibition of TGF-β signaling transduction. TGF-β is a
multifunctional cytokine that induces epithelial-mesenchymal transition (EMT) of cancer cells, and it
is also responsible for the maintenance of EMT, a critical event during early metastatic growth. The
mechanism by which EGCG modulates TGF-β pathway has not been completely elucidated. It has
been shown that the binding between TGF-β and its receptor, TGFR-II, activates two different
pathways leading to EMT: the canonical Smad-dependent pathway and the mitogen-activated
protein kinase (MAPK) pathway. Tabuchi et al. used immunoprecipitation and affinity
chromatography assays to demonstrate binding between EGCG and TGFR-II protein. This
interaction may be responsible for the inhibitory effect of EGCG on the expression of alpha-SMA
(considered a marker of the EMT) via the TGF-beta Smad2/3 pathway in human lung fibroblast cells
[94].
EGCG has also been shown to bind to metalloproteinases (MMPs). MMPs are matrix degrading
enzymes that are involved in tumor invasion and metastasis [50] whose expression is regulated by
several growth factors, including TGF-β1 [216–219]. Sazuka et al. have demonstrated that EGCG
inhibits the collagenase activity of MMP-2 and MMP-9 produced by lung carcinoma cells. The
authors suggest that the mechanism of inhibition relies on direct binding between EGCG and MMP
Nutrients 2018, 10, 1936 11 of 24
proteins, as proved by affinity gel chromatography experiments [50]. In 2017, Chowdhury et al.
performed a preliminary in silico analysis and then showed a strong interaction of pro-/active
MMP2 with the galloyl group of EGCG and ECG in pulmonary artery smooth muscle cell culture
supernatant. They showed that EGCG and ECG were better inhibitors of proMMP2 when compared
to MMP2, and they demonstrated that a strong interaction with MT1/MMP is involved in the
conversion of proMMP2 to active MMP2 [220]. Further, investigating the interactions of pro-/active
MMP-9 with green tea catechins by computational methods, they showed strong interactions
between pro-/active MMP9 and EGCG/ECG [221].
4. EGCG Epigenetic Regulation
Another mechanism that can explain the pleiotropic effects exerted by green tea catechins in
tumor cells is the epigenetic change in gene expression and chromatin organization. Mutations in
oncogenes and tumor suppressor genes are often the cause of cancer development and alterations of
gene expression count for cancer progression.
Many biologically active compounds, including EGCG, have been demonstrated to modulate
DNA methylation and histone acetylation status [222].
DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) are enzymes that are
involved in transcriptional gene silencing and histone acetyl transferases (HATs) positively regulate
gene expression regulation [223,224]. Several studies reported EGCG contribution in epigenetic
control acting on DNMTs, HDACs and HATs expression and activity in different tumours. We will
briefly mention different genes whose expression is enhanced or reduced by EGCG-dependent
epigenetic control.
Fang et al. demonstrated that EGCG binds to DNMT and competitively inhibits the enzymatic
activity (Ki of 6.89 µM), yielding the reactivation of methylation-silenced genes in prostate cancer
PC3 cells [225]. Molecular modeling and docking studies supported the binding of EGCG to
DNMT3B and HDAC1 [39].
In HeLa cell line, it has been observed that EGCG can direct bind to and inhibit DNMT1,
DNMT3B, and HDAC1 activity, causing a reduction in DNA hypermethylation and restoring the
expression of repressed genes as retinoic acid receptor β (RARβ), CDH1 (e-cadherine gene), and
DAPK1 [29,39]. Furthermore, in the same the same cell line, EGCG combination with
eugenol-amrogentin (active compounds of clove and Swertia Chirata, respectively) reduces DNMT1
expression with the consequent hypomethylation of the cell cycle inhibitors p16 and LimD1
promoters [226]. In acute promyelocytic leukemia cells, EGCG down-regulates DNMT1, HDAC1,
HDAC2, G9a, and Polycomb repressive complex 2 (PRC2) core components expression and favours
the binding of hyperacetylated H4 and acetylated H3K14 histones to promoter regions of p27, CAF,
C/EBPα, and C/EBPε genes [63]. In the lung cell line PC-9, EGCG combination with Am80 (a
synthetic retinoid used for acute promyelocytic leukemia therapy) causes a decrease of HDAC4,
HDAC5, and HDAC6 protein levels and reduction of HDAC activity, leading to increased p53 and
α-tubulin acetylation [227]. In in vitro and in vivo models of lung cancer, EGCG has been found to
resensitize tumor cells to Cisplatin (DDP)-based combination chemotherapy through DNMT and
HDAC activity inhibition, and the subsequent re-expression of GAS1, TIMP4, ICAM1, and WISP2
genes [228]. In in vivo model of lung cancer, EGCG epigenetic action in down-regulating DNMT1 is
accompanied by phospho-histone H2AX (γ-H2AX) and p-AKT reduction [229]. In skin cancer cells,
it has demonstrated EGCG capability in reducing DNMT1, DNMT3A, and DNMT3B activity and
expression, and also in increasing histones H3 and H4 acetylation. As a consequence of the described
epigenetic changes, a restored expression of the cell cycle inhibitors p16 and p21 has been observed
[48]. In breast cancer cells, EGCG-dependent reduction of HDAC1 and zeste homolog 2 (EZH2)
protein levels leads to tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) gene transcriptional
activation [72]. In prostate cancer cell lines, it has been observed that the EGCG-dependent reduction
of the acetylated androgen receptor (AR) gene might be induced by EGCG reduction in HAT activity
[65]. EGCG also acts on teleomerase, reducing its activity in different tumor types as esophageal
carcinoma [230], glioma [231], cervical cancer [232], breast cancer [100], nasopharyngeal carcinoma
Nutrients 2018, 10, 1936 12 of 24
[233], ovarian cancer [68], laryngeal squamous cell carcinoma [234], and lung cancer [235]. It has also
been shown that EGCG can translocate from the cytoplasm to the nucleus where it can bind to DNA,
suggesting a possible role in gene expression regulation also through the direct binding to nucleic
acid [236,237]. However, the effects of EGCG/DNA direct interaction need to be clarified.
5. Conclusions
Because of their anti-proliferative, pro-apoptotic, and anti-oxidative properties, green tea
catechins and especially EGCG are receiving much attention in cancer biology. Several in vitro, in
vivo, and clinical studies, have demonstrated that EGCG exerts anti-cancer effects in different
models through the activation/inhibition of several signalling pathways, most of which are triggered
by the direct interaction between EGCG and specific protein targets. The array of EGCG interactors
is wide and growing, and it includes intracellular molecules, membranes receptors, membrane
microdomains, and the plasma membrane itself. One of the first EGCG direct target identified was
67LR, but in recent years, others interactors, such as Pin1 or TGFR-II, have been recognized.
Appropriate identification and study of EGCG direct targets will allow a better understanding of its
mechanisms of action and a better exploitation of its anti-cancer properties. From 2004, when the
67LR was first identified as direct target of EGCG by Tachibana et al., several research teams have
investigated the pathways modulated by EGCG-67LR interaction. Today, we know that the
anti-proliferative action of EGCG is mediated by the binding to 67LR, whose expression is increased
in tumour cells. Convincing experimental data also showed that membrane composition is involved
in the inhibitory activity of EGCG in some cancer cells lines. Since 67LR is generally located in lipid
rafts, EGCG-mediated microdomains composition and the alteration of their functions triggers the
downstream signalling cascades. In addition, new experimental data have brought to light novel
EGCG signalling cascades leading to cell apoptosis, cell cycle arrest, reduction in CSC colony and
spheroid formation, as well as regulation of miRNAs expression. EGCG binding to membrane
receptors, such as TGFR-II, intracellular molecules, such as Pin1 and secreted enzymes, such as
MMPs, provided noteworthy information about the mechanisms of EGCG-mediated tumour
suppression. Another mechanism to explain the pleiotropic anti-cancer effects that are exerted by
EGCG and green tea catechins that is gaining the attention of the researchers is the modulation of
epigenetic processes. Long-term administration of green tea catechins leads to the re-activation of
tumour suppressor genes that are silenced during carcinogenesis and downregulation of oncogenes
through the inhibition of enzymes, such as DNMTs and HDACs involved in DNA methylation and
chromatin remodelling. Further studies on the interaction of EGCG with protein targets will provide
new insights enabling the development of more pharmacological treatments targeting
EGCG-activated master regulators of key pathways.
Author Contributions: A.N., V.N., F.R. and S.B. critically reviewed the literature and wrote the manuscript.
Funding: This research received no external funding.
Acknowledgments: V.N. was supported by a fellowship by Fondazione Umberto Veronesi. We thank Paul
Wegener for English editing support.
Conflicts of Interest: The authors declare no conflict of interest.
References
1. Graham, H.N. Green tea composition, consumption, and polyphenol chemistry. Prev. Med. 1992, 21, 334–
350.
2. Lowe, G.M.; Gana, K.; Rahman, K. Dietary supplementation with green tea extract promotes enhanced
human leukocyte activity. J. Complement. Integr. Med. 2015, 12, 277–282.
3. Boschmann, M.; Thielecke, F. The effects of epigallocatechin-3-gallate on thermogenesis and fat oxidation
in obese men: A pilot study. J. Am. Coll. Nutr. 2007, 26, 389S–395S.
Nutrients 2018, 10, 1936 13 of 24
4. Basu, A.; Sanchez, K.; Leyva, M.J.; Wu, M.; Betts, N.M.; Aston, C.E.; Lyons, T.J. Green tea supplementation
affects body weight, lipids, and lipid peroxidation in obese subjects with metabolic syndrome. J. Am. Coll.
Nutr. 2010, 29, 31–40.
5. Ide, K.; Yamada, H.; Takuma, N.; Park, M.; Wakamiya, N.; Nakase, J.; Ukawa, Y.; Sagesaka, Y.M. Green tea
consumption affects cognitive dysfunction in the elderly: A pilot study. Nutrients 2014, 6, 4032–4042.
6. Bettuzzi, S.; Brausi, M.; Rizzi, F.; Castagnetti, G.; Peracchia, G.; Corti, A. Chemoprevention of human
prostate cancer by oral administration of green tea catechins in volunteers with high-grade prostate
intraepithelial neoplasia: A preliminary report from a one-year proof-of-principle study. Cancer Res. 2006,
66, 1234–1240.
7. Khan, N.; Afaq, F.; Saleem, M.; Ahmad, N.; Mukhtar, H. Targeting multiple signaling pathways by green
tea polyphenol (-)-epigallocatechin-3-gallate. Cancer Res. 2006, 66, 2500–2505.
8. Yang, C.S.; Sang, S.; Lambert, J.D.; Lee, M.J. Bioavailability issues in studying the health effects of plant
polyphenolic compounds. Mol. Nutr. Food Res. 2008, 52 (Suppl. 1), S139–S151.
9. Lee, M.J.; Wang, Z.Y.; Li, H.; Chen, L.; Sun, Y.; Gobbo, S.; Balentine, D.A.; Yang, C.S. Analysis of plasma
and urinary tea polyphenols in human subjects. Cancer Epidemiol. Biomarkers Prev. 1995, 4, 393–399.
10. Gan, R.Y.; Li, H.B.; Sui, Z.Q.; Corke, H. Absorption, metabolism, anti-cancer effect and molecular targets of
epigallocatechin gallate (egcg): An updated review. Crit. Rev. Food Sci. Nutr. 2018, 58, 924–941.
11. Zubair, H.; Azim, S.; Ahmad, A.; Khan, M.A.; Patel, G.K.; Singh, S.; Singh, A.P. Cancer chemoprevention
by phytochemicals: Nature’s healing touch. Molecules 2017, 22, 395.
12. Jin, G.; Yang, Y.; Liu, K.; Zhao, J.; Chen, X.; Liu, H.; Bai, R.; Li, X.; Jiang, Y.; Zhang, X.; et al. Combination
curcumin and (-)-epigallocatechin-3-gallate inhibits colorectal carcinoma microenvironment-induced
angiogenesis by jak/stat3/il-8 pathway. Oncogenesis 2017, 6, e384.
13. Chan, M.M.; Chen, R.; Fong, D. Targeting cancer stem cells with dietary phytochemical—Repositioned
drug combinations. Cancer Lett. 2018, 433, 53–64.
14. Wang, W.; Chen, D.; Zhu, K. Sox2ot variant 7 contributes to the synergistic interaction between egcg and
doxorubicin to kill osteosarcoma via autophagy and stemness inhibition. J. Exp. Clin. Cancer Res. 2018, 37,
37.
15. Mayr, C.; Wagner, A.; Neureiter, D.; Pichler, M.; Jakab, M.; Illig, R.; Berr, F.; Kiesslich, T. The green tea
catechin epigallocatechin gallate induces cell cycle arrest and shows potential synergism with cisplatin in
biliary tract cancer cells. BMC Complement. Altern. Med. 2015, 15, 194.
16. Zhou, Y.; Tang, J.; Du, Y.; Ding, J.; Liu, J.Y. The green tea polyphenol egcg potentiates the antiproliferative
activity of sunitinib in human cancer cells. Tumour. Biol. 2016, 37, 8555–8566.
17. Yuan, X.; He, Y.; Zhou, G.; Li, X.; Feng, A.; Zheng, W. Target challenging-cancer drug delivery to gastric
cancer tissues with a fucose graft epigallocatechin-3-gallate-gold particles nanocomposite approach. J.
Photochem. Photobiol. B 2018, 183, 147–153.
18. Hajipour, H.; Hamishehkar, H.; Nazari Soltan Ahmad, S.; Barghi, S.; Maroufi, N.F.; Taheri, R.A. Improved
anticancer effects of epigallocatechin gallate using rgd-containing nanostructured lipid carriers. Artif. Cells
Nanomed. Biotechnol. 2018, doi:10.1080/21691401.2017.1423493.
19. Sanna, V.; Singh, C.K.; Jashari, R.; Adhami, V.M.; Chamcheu, J.C.; Rady, I.; Sechi, M.; Mukhtar, H.;
Siddiqui, I.A. Targeted nanoparticles encapsulating (-)-epigallocatechin-3-gallate for prostate cancer
prevention and therapy. Sci. Rep. 2017, 7, 41573.
20. Krupkova, O.; Ferguson, S.J.; Wuertz-Kozak, K. Stability of (-)-epigallocatechin gallate and its activity in
liquid formulations and delivery systems. J. Nutr. Biochem. 2016, 37, 1–12.
21. Rizzi, F.; Naponelli, V.; Silva, A.; Modernelli, A.; Ramazzina, I.; Bonacini, M.; Tardito, S.; Gatti, R.; Uggeri,
J.; Bettuzzi, S. Polyphenon e(r), a standardized green tea extract, induces endoplasmic reticulum stress,
leading to death of immortalized pnt1a cells by anoikis and tumorigenic pc3 by necroptosis. Carcinogenesis
2014, 35, 828–839.
22. Modernelli, A.; Naponelli, V.; Giovanna Troglio, M.; Bonacini, M.; Ramazzina, I.; Bettuzzi, S.; Rizzi, F.
Egcg antagonizes bortezomib cytotoxicity in prostate cancer cells by an autophagic mechanism. Sci. Rep.
2015, 5, 15270.
23. Thawonsuwan, J.; Kiron, V.; Satoh, S.; Panigrahi, A.; Verlhac, V. Epigallocatechin-3-gallate (egcg) affects
the antioxidant and immune defense of the rainbow trout, oncorhynchus mykiss. Fish. Physiol. Biochem.
2010, 36, 687–697.
Nutrients 2018, 10, 1936 14 of 24
24. Lambert, J.D.; Elias, R.J. The antioxidant and pro-oxidant activities of green tea polyphenols: A role in
cancer prevention. Arch. Biochem. Biophys. 2010, 501, 65–72.
25. Gupta, S.; Hastak, K.; Afaq, F.; Ahmad, N.; Mukhtar, H. Essential role of caspases in
epigallocatechin-3-gallate-mediated inhibition of nuclear factor kappa b and induction of apoptosis.
Oncogene 2004, 23, 2507–2522.
26. Shimizu, M.; Adachi, S.; Masuda, M.; Kozawa, O.; Moriwaki, H. Cancer chemoprevention with green tea
catechins by targeting receptor tyrosine kinases. Mol. Nutr. Food Res. 2011, 55, 832–843.
27. Singh, B.N.; Shankar, S.; Srivastava, R.K. Green tea catechin, epigallocatechin-3-gallate (egcg):
Mechanisms, perspectives and clinical applications. Biochem. Pharmacol. 2011, 82, 1807–1821.
28. Lin, C.H.; Shen, Y.A.; Hung, P.H.; Yu, Y.B.; Chen, Y.J. Epigallocathechin gallate, polyphenol present in
green tea, inhibits stem-like characteristics and epithelial-mesenchymal transition in nasopharyngeal
cancer cell lines. BMC Complement. Altern. Med. 2012, 12, 201.
29. Lee, W.J.; Shim, J.Y.; Zhu, B.T. Mechanisms for the inhibition of DNA methyltransferases by tea catechins
and bioflavonoids. Mol. Pharmacol. 2005, 68, 1018–1030.
30. Shirakami, Y.; Shimizu, M. Possible mechanisms of green tea and its constituents against cancer. Molecules
2018, 23, 2284.
31. Rahmani, A.H.; Al Shabrmi, F.M.; Allemailem, K.S.; Aly, S.M.; Khan, M.A. Implications of green tea and
its constituents in the prevention of cancer via the modulation of cell signalling pathway. Biomed. Res. Int.
2015, 2015, 925640.
32. Naponelli, V.; Ramazzina, I.; Lenzi, C.; Bettuzzi, S.; Rizzi, F. Green tea catechins for prostate cancer
prevention: Present achievements and future challenges. Antioxidants 2017, 6, 26.
33. Ellinger, S.; Muller, N.; Stehle, P.; Ulrich-Merzenich, G. Consumption of green tea or green tea products: Is
there an evidence for antioxidant effects from controlled interventional studies? Phytomedicine 2011, 18,
903–915.
34. Shankar, S.; Ganapathy, S.; Hingorani, S.R.; Srivastava, R.K. Egcg inhibits growth, invasion, angiogenesis
and metastasis of pancreatic cancer. Front. Biosci. 2008, 13, 440–452.
35. Gupta, S.; Hussain, T.; Mukhtar, H. Molecular pathway for (-)-epigallocatechin-3-gallate-induced cell cycle
arrest and apoptosis of human prostate carcinoma cells. Arch. Biochem. Biophys. 2003, 410, 177–185.
36. Shimizu, M.; Shirakami, Y.; Moriwaki, H. Targeting receptor tyrosine kinases for chemoprevention by
green tea catechin, egcg. Int. J. Mol. Sci. 2008, 9, 1034–1049.
37. Shimizu, M.; Weinstein, I.B. Modulation of signal transduction by tea catechins and related
phytochemicals. Mutat. Res. 2005, 591, 147–160.
38. Pandey, M.; Shukla, S.; Gupta, S. Promoter demethylation and chromatin remodeling by green tea
polyphenols leads to re-expression of gstp1 in human prostate cancer cells. Int. J. Cancer 2010, 126, 2520–
2533.
39. Khan, M.A.; Hussain, A.; Sundaram, M.K.; Alalami, U.; Gunasekera, D.; Ramesh, L.; Hamza, A.; Quraishi,
U. (-)-epigallocatechin-3-gallate reverses the expression of various tumor-suppressor genes by inhibiting
DNA methyltransferases and histone deacetylases in human cervical cancer cells. Oncol. Rep. 2015, 33,
1976–1984.
40. Rouzer, C.A.; Marnett, L.J. Green tea gets molecular. Cancer Prev. Res. 2011, 4, 1343–1345.
41. Saeki, K.; Hayakawa, S.; Nakano, S.; Ito, S.; Oishi, Y.; Suzuki, Y.; Isemura, M. In vitro and in silico studies
of the molecular interactions of epigallocatechin-3-o-gallate (egcg) with proteins that explain the health
benefits of green tea. Molecules 2018, 23, 1295.
42. Ermakova, S.; Choi, B.Y.; Choi, H.S.; Kang, B.S.; Bode, A.M.; Dong, Z. The intermediate filament protein
vimentin is a new target for epigallocatechin gallate. J. Biol. Chem. 2005, 280, 16882–16890.
43. He, Z.; Tang, F.; Ermakova, S.; Li, M.; Zhao, Q.; Cho, Y.Y.; Ma, W.Y.; Choi, H.S.; Bode, A.M.; Yang, C.S.; et
al. Fyn is a novel target of (-)-epigallocatechin gallate in the inhibition of jb6 cl41 cell transformation. Mol.
Carcinog. 2008, 47, 172–183.
44. Shim, J.H.; Choi, H.S.; Pugliese, A.; Lee, S.Y.; Chae, J.I.; Choi, B.Y.; Bode, A.M.; Dong, Z.
(-)-epigallocatechin gallate regulates cd3-mediated t cell receptor signaling in leukemia through the
inhibition of zap-70 kinase. J. Biol. Chem. 2008, 283, 28370–28379.
45. Li, M.; He, Z.; Ermakova, S.; Zheng, D.; Tang, F.; Cho, Y.Y.; Zhu, F.; Ma, W.Y.; Sham, Y.; Rogozin, E.A.; et
al. Direct inhibition of insulin-like growth factor-i receptor kinase activity by (-)-epigallocatechin-3-gallate
regulates cell transformation. Cancer Epidemiol. Biomarkers Prev. 2007, 16, 598–605.
Nutrients 2018, 10, 1936 15 of 24
46. Ermakova, S.P.; Kang, B.S.; Choi, B.Y.; Choi, H.S.; Schuster, T.F.; Ma, W.Y.; Bode, A.M.; Dong, Z.
(-)-epigallocatechin gallate overcomes resistance to etoposide-induced cell death by targeting the
molecular chaperone glucose-regulated protein 78. Cancer Res. 2006, 66, 9260–9269.
47. Fujimura, Y. Small molecule-sensing strategy and techniques for understanding the functionality of green
tea. Biosci. Biotechnol. Biochem. 2015, 79, 687–699.
48. Nandakumar, V.; Vaid, M.; Katiyar, S.K. (-)-epigallocatechin-3-gallate reactivates silenced tumor
suppressor genes, cip1/p21 and p16ink4a, by reducing DNA methylation and increasing histones
acetylation in human skin cancer cells. Carcinogenesis 2011, 32, 537–544.
49. Hu, Q.; Chang, X.; Yan, R.; Rong, C.; Yang, C.; Cheng, S.; Gu, X.; Yao, H.; Hou, X.; Mo, Y.; et al.
(-)-epigallocatechin-3-gallate induces cancer cell apoptosis via acetylation of amyloid precursor protein.
Med. Oncol. 2015, 32, 390.
50. Sazuka, M.; Imazawa, H.; Shoji, Y.; Mita, T.; Hara, Y.; Isemura, M. Inhibition of collagenases from mouse
lung carcinoma cells by green tea catechins and black tea theaflavins. Biosci. Biotechnol. Biochem. 1997, 61,
1504–1506.
51. Fujimura, Y.; Tachibana, H.; Yamada, K. Lipid raft-associated catechin suppresses the fcepsilonri
expression by inhibiting phosphorylation of the extracellular signal-regulated kinase1/2. FEBS Lett. 2004,
556, 204–210.
52. Tsukamoto, S.; Huang, Y.; Kumazoe, M.; Lesnick, C.; Yamada, S.; Ueda, N.; Suzuki, T.; Yamashita, S.; Kim,
Y.H.; Fujimura, Y.; et al. Sphingosine kinase-1 protects multiple myeloma from apoptosis driven by
cancer-specific inhibition of rtks. Mol. Cancer Ther. 2015, 14, 2303–2312.
53. Wu, P.P.; Kuo, S.C.; Huang, W.W.; Yang, J.S.; Lai, K.C.; Chen, H.J.; Lin, K.L.; Chiu, Y.J.; Huang, L.J.;
Chung, J.G. (-)-epigallocatechin gallate induced apoptosis in human adrenal cancer nci-h295 cells through
caspase-dependent and caspase-independent pathway. Anticancer Res. 2009, 29, 1435–1442.
54. Zhu, J.; Jiang, Y.; Yang, X.; Wang, S.; Xie, C.; Li, X.; Li, Y.; Chen, Y.; Wang, X.; Meng, Y.; et al.
Wnt/beta-catenin pathway mediates (-)-epigallocatechin-3-gallate (egcg) inhibition of lung cancer stem
cells. Biochem. Biophys. Res. Commun. 2017, 482, 15–21.
55. Umeda, D.; Tachibana, H.; Yamada, K. Epigallocatechin-3-o-gallate disrupts stress fibers and the
contractile ring by reducing myosin regulatory light chain phosphorylation mediated through the target
molecule 67 kda laminin receptor. Biochem. Biophys. Res. Commun. 2005, 333, 628–635.
56. Shankar, S.; Suthakar, G.; Srivastava, R.K. Epigallocatechin-3-gallate inhibits cell cycle and induces
apoptosis in pancreatic cancer. Front. Biosci. 2007, 12, 5039–5051.
57. Hwang, Y.S.; Park, K.K.; Chung, W.Y. Epigallocatechin-3 gallate inhibits cancer invasion by repressing
functional invadopodia formation in oral squamous cell carcinoma. Eur. J. Pharmacol. 2013, 715, 286–295.
58. Olotu, F.A.; Agoni, C.; Adeniji, E.; Abdullahi, M.; Soliman, M.E. Probing gallate-mediated selectivity and
high-affinity binding of epigallocatechin gallate: A way-forward in the design of selective inhibitors for
anti-apoptotic bcl-2 proteins. Appl. Biochem. Biotechnol. 2018, 29, 1–20.
59. Shin, S.; Kim, M.K.; Jung, W.; Chong, Y. (-)-epigallocatechin gallate derivatives reduce the expression of
both urokinase plasminogen activator and plasminogen activator inhibitor-1 to inhibit migration,
adhesion, and invasion of mda-mb-231 cells. Phytother. Res. 2018, 32, 2086–2096.
60. Shi, J.; Liu, F.; Zhang, W.; Liu, X.; Lin, B.; Tang, X. Epigallocatechin-3-gallate inhibits nicotine-induced
migration and invasion by the suppression of angiogenesis and epithelial-mesenchymal transition in
non-small cell lung cancer cells. Oncol. Rep. 2015, 33, 2972–2980.
61. Umeda, D.; Yano, S.; Yamada, K.; Tachibana, H. Green tea polyphenol epigallocatechin-3-gallate signaling
pathway through 67-kda laminin receptor. J. Biol. Chem. 2008, 283, 3050–3058.
62. Jang, J.Y.; Lee, J.K.; Jeon, Y.K.; Kim, C.W. Exosome derived from epigallocatechin gallate treated breast
cancer cells suppresses tumor growth by inhibiting tumor-associated macrophage infiltration and m2
polarization. BMC Cancer 2013, 13, 421.
63. Borutinskaite, V.; Virksaite, A.; Gudelyte, G.; Navakauskiene, R. Green tea polyphenol egcg causes
anti-cancerous epigenetic modulations in acute promyelocytic leukemia cells. Leuk. Lymphoma 2018, 59,
469–478.
64. Ma, J.; Shi, M.; Li, G.; Wang, N.; Wei, J.; Wang, T.; Wang, Y. Regulation of id1 expression by
epigallocatechin3gallate and its effect on the proliferation and apoptosis of poorly differentiated ags
gastric cancer cells. Int. J. Oncol. 2013, 43, 1052–1058.
Nutrients 2018, 10, 1936 16 of 24
65. Lee, Y.H.; Kwak, J.; Choi, H.K.; Choi, K.C.; Kim, S.; Lee, J.; Jun, W.; Park, H.J.; Yoon, H.G. Egcg suppresses
prostate cancer cell growth modulating acetylation of androgen receptor by anti-histone acetyltransferase
activity. Int. J. Mol. Med. 2012, 30, 69–74.
66. Balasubramanian, S.; Adhikary, G.; Eckert, R.L. The bmi-1 polycomb protein antagonizes the
(-)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival. Carcinogenesis 2010, 31,
496–503.
67. Takahashi, A.; Watanabe, T.; Mondal, A.; Suzuki, K.; Kurusu-Kanno, M.; Li, Z.; Yamazaki, T.; Fujiki, H.;
Suganuma, M. Mechanism-based inhibition of cancer metastasis with (-)-epigallocatechin gallate. Biochem.
Biophys. Res. Commun. 2014, 443, 1–6.
68. Chen, H.; Landen, C.N.; Li, Y.; Alvarez, R.D.; Tollefsbol, T.O. Epigallocatechin gallate and sulforaphane
combination treatment induce apoptosis in paclitaxel-resistant ovarian cancer cells through htert and bcl-2
down-regulation. Exp. Cell Res. 2013, 319, 697–706.
69. Basu, A.; Haldar, S. Combinatorial effect of epigallocatechin-3-gallate and trail on pancreatic cancer cell
death. Int. J. Oncol. 2009, 34, 281–286.
70. Belguise, K.; Guo, S.; Sonenshein, G.E. Activation of foxo3a by the green tea polyphenol
epigallocatechin-3-gallate induces estrogen receptor alpha expression reversing invasive phenotype of
breast cancer cells. Cancer Res. 2007, 67, 5763–5770.
71. Harper, C.E.; Patel, B.B.; Wang, J.; Eltoum, I.A.; Lamartiniere, C.A. Epigallocatechin-3-gallate suppresses
early stage, but not late stage prostate cancer in tramp mice: Mechanisms of action. Prostate 2007, 67, 1576–
1589.
72. Deb, G.; Thakur, V.S.; Limaye, A.M.; Gupta, S. Epigenetic induction of tissue inhibitor of matrix
metalloproteinase-3 by green tea polyphenols in breast cancer cells. Mol. Carcinog. 2015, 54, 485–499.
73. Moses, M.A.; Henry, E.C.; Ricke, W.A.; Gasiewicz, T.A. The heat shock protein 90 inhibitor,
(-)-epigallocatechin gallate, has anticancer activity in a novel human prostate cancer progression model.
Cancer Prev. Res. 2015, 8, 249–257.
74. Kumazoe, M.; Sugihara, K.; Tsukamoto, S.; Huang, Y.; Tsurudome, Y.; Suzuki, T.; Suemasu, Y.; Ueda, N.;
Yamashita, S.; Kim, Y.; et al. 67-kda laminin receptor increases cgmp to induce cancer-selective apoptosis.
J. Clin. Investig. 2013, 123, 787–799.
75. Vahora, H.; Khan, M.A.; Alalami, U.; Hussain, A. The potential role of nitric oxide in halting cancer
progression through chemoprevention. J. Cancer Prev. 2016, 21, 1–12.
76. Surh, Y.J.; Chun, K.S.; Cha, H.H.; Han, S.S.; Keum, Y.S.; Park, K.K.; Lee, S.S. Molecular mechanisms
underlying chemopreventive activities of anti-inflammatory phytochemicals: Down-regulation of cox-2
and inos through suppression of nf-kappa b activation. Mutat. Res. 2001, 480–481, 243–268.
77. Hayakawa, S.; Saito, K.; Miyoshi, N.; Ohishi, T.; Oishi, Y.; Miyoshi, M.; Nakamura, Y. Anti-cancer effects of
green tea by either anti- or pro- oxidative mechanisms. Asian Pac. J. Cancer Prev. 2016, 17, 1649–1654.
78. Dhakshinamoorthy, S.; Porter, A.G. Nitric oxide-induced transcriptional up-regulation of protective genes
by nrf2 via the antioxidant response element counteracts apoptosis of neuroblastoma cells. J. Biol. Chem.
2004, 279, 20096–20107.
79. Li, Y.J.; Wu, S.L.; Lu, S.M.; Chen, F.; Guo, Y.; Gan, S.M.; Shi, Y.L.; Liu, S.; Li, S.L.
(-)-epigallocatechin-3-gallate inhibits nasopharyngeal cancer stem cell self-renewal and migration and
reverses the epithelial-mesenchymal transition via nf-kappab p65 inactivation. Tumour. Biol. 2015, 36,
2747–2761.
80. He, L.; Zhang, E.; Shi, J.; Li, X.; Zhou, K.; Zhang, Q.; Le, A.D.; Tang, X. (-)-epigallocatechin-3-gallate
inhibits human papillomavirus (hpv)-16 oncoprotein-induced angiogenesis in non-small cell lung cancer
cells by targeting hif-1alpha. Cancer Chemother. Pharmacol. 2013, 71, 713–725.
81. Yamada, S.; Tsukamoto, S.; Huang, Y.; Makio, A.; Kumazoe, M.; Yamashita, S.; Tachibana, H.
Epigallocatechin-3-O-gallate up-regulates microrna-let-7b expression by activating 67-kda laminin
receptor signaling in melanoma cells. Sci. Rep. 2016, 6, 19225.
82. Wang, H.; Bian, S.; Yang, C.S. Green tea polyphenol egcg suppresses lung cancer cell growth through
upregulating mir-210 expression caused by stabilizing hif-1alpha. Carcinogenesis 2011, 32, 1881–1889.
83. Urusova, D.V.; Shim, J.H.; Kim, D.J.; Jung, S.K.; Zykova, T.A.; Carper, A.; Bode, A.M.; Dong, Z.
Epigallocatechin-gallate suppresses tumorigenesis by directly targeting pin1. Cancer Prev. Res. 2011, 4,
1366–1377.
Nutrients 2018, 10, 1936 17 of 24
84. Qin, J.; Chen, H.G.; Yan, Q.; Deng, M.; Liu, J.; Doerge, S.; Ma, W.; Dong, Z.; Li, D.W. Protein
phosphatase-2a is a target of epigallocatechin-3-gallate and modulates p53-bak apoptotic pathway. Cancer
Res. 2008, 68, 4150–4162.
85. Toden, S.; Tran, H.M.; Tovar-Camargo, O.A.; Okugawa, Y.; Goel, A. Epigallocatechin-3-gallate targets
cancer stem-like cells and enhances 5-fluorouracil chemosensitivity in colorectal cancer. Oncotarget 2016, 7,
16158–16171.
86. Tsukamoto, S.; Huang, Y.; Umeda, D.; Yamada, S.; Yamashita, S.; Kumazoe, M.; Kim, Y.; Murata, M.;
Yamada, K.; Tachibana, H. 67-kda laminin receptor-dependent protein phosphatase 2a (pp2a) activation
elicits melanoma-specific antitumor activity overcoming drug resistance. J. Biol. Chem. 2014, 289, 32671–
32681.
87. Saldanha, S.N.; Kala, R.; Tollefsbol, T.O. Molecular mechanisms for inhibition of colon cancer cells by
combined epigenetic-modulating epigallocatechin gallate and sodium butyrate. Exp. Cell Res. 2014, 324,
40–53.
88. Zhang, Y.; Wang, S.X.; Ma, J.W.; Li, H.Y.; Ye, J.C.; Xie, S.M.; Du, B.; Zhong, X.Y. Egcg inhibits properties of
glioma stem-like cells and synergizes with temozolomide through downregulation of p-glycoprotein
inhibition. J. Neurooncol. 2015, 121, 41–52.
89. Suzuki, Y.; Isemura, M. Binding interaction between (-)-epigallocatechin gallate causes impaired
spreading of cancer cells on fibrinogen. Biomed. Res. 2013, 34, 301–308.
90. Tachibana, H.; Koga, K.; Fujimura, Y.; Yamada, K. A receptor for green tea polyphenol egcg. Nat. Struct.
Mol. Biol. 2004, 11, 380–381.
91. Lee, S.H.; Nam, H.J.; Kang, H.J.; Kwon, H.W.; Lim, Y.C. Epigallocatechin-3-gallate attenuates head and
neck cancer stem cell traits through suppression of notch pathway. Eur. J. Cancer 2013, 49, 3210–3218.
92. Adachi, S.; Nagao, T.; Ingolfsson, H.I.; Maxfield, F.R.; Andersen, O.S.; Kopelovich, L.; Weinstein, I.B. The
inhibitory effect of (-)-epigallocatechin gallate on activation of the epidermal growth factor receptor is
associated with altered lipid order in ht29 colon cancer cells. Cancer Res. 2007, 67, 6493–6501.
93. Duhon, D.; Bigelow, R.L.; Coleman, D.T.; Steffan, J.J.; Yu, C.; Langston, W.; Kevil, C.G.; Cardelli, J.A. The
polyphenol epigallocatechin-3-gallate affects lipid rafts to block activation of the c-met receptor in prostate
cancer cells. Mol. Carcinog. 2010, 49, 739–749.
94. Tabuchi, M.; Hayakawa, S.; Honda, E.; Ooshima, K.; Itoh, T.; Yoshida, K.; Park, A.M.; Higashino, H.;
Isemura, M.; Munakata, H. Epigallocatechin-3-gallate suppresses transforming growth factor-beta
signaling by interacting with the transforming growth factor-beta type ii receptor. World J. Exp. Med. 2013,
3, 100–107.
95. Wubetu, G.Y.; Shimada, M.; Morine, Y.; Ikemoto, T.; Ishikawa, D.; Iwahashi, S.; Yamada, S.; Saito, Y.;
Arakawa, Y.; Imura, S. Epigallocatechin gallate hinders human hepatoma and colon cancer sphere
formation. J. Gastroenterol. Hepatol. 2016, 31, 256–264.
96. Punathil, T.; Tollefsbol, T.O.; Katiyar, S.K. Egcg inhibits mammary cancer cell migration through
inhibition of nitric oxide synthase and guanylate cyclase. Biochem. Biophys. Res. Commun. 2008, 375, 162–
167.
97. Sen, T.; Dutta, A.; Chatterjee, A. Epigallocatechin-3-gallate (egcg) downregulates gelatinase-b (mmp-9) by
involvement of fak/erk/nfkappab and ap-1 in the human breast cancer cell line mda-mb-231. Anticancer
Drugs 2010, 21, 632–644.
98. Singh, T.; Katiyar, S.K. Green tea polyphenol, (-)-epigallocatechin-3-gallate, induces toxicity in human skin
cancer cells by targeting beta-catenin signaling. Toxicol. Appl. Pharmacol. 2013, 273, 418–424.
99. Shankar, S.; Marsh, L.; Srivastava, R.K. Egcg inhibits growth of human pancreatic tumors orthotopically
implanted in balb c nude mice through modulation of fkhrl1/foxo3a and neuropilin. Mol. Cell. Biochem.
2013, 372, 83–94.
100. Moradzadeh, M.; Hosseini, A.; Erfanian, S.; Rezaei, H. Epigallocatechin-3-gallate promotes apoptosis in
human breast cancer t47d cells through down-regulation of pi3k/akt and telomerase. Pharmacol. Rep. 2017,
69, 924–928.
101. Ketchart, W.; Smith, K.M.; Krupka, T.; Wittmann, B.M.; Hu, Y.; Rayman, P.A.; Doughman, Y.Q.; Albert,
J.M.; Bai, X.; Finke, J.H.; et al. Inhibition of metastasis by hexim1 through effects on cell invasion and
angiogenesis. Oncogene 2013, 32, 3829–3839.
Nutrients 2018, 10, 1936 18 of 24
102. Lu, C.L.; Xu, J.; Yao, H.J.; Luo, K.L.; Li, J.M.; Wu, T.; Wu, G.Z. Inhibition of human 67-kda laminin receptor
sensitizes multidrug resistance colon cancer cell line sw480 for apoptosis induction. Tumour. Biol. 2016, 37,
1319–1325.
103. Montuori, N.; Selleri, C.; Risitano, A.M.; Raiola, A.M.; Ragno, P.; Del Vecchio, L.; Rotoli, B.; Rossi, G.
Expression of the 67-kda laminin receptor in acute myeloid leukemia cells mediates adhesion to laminin
and is frequently associated with monocytic differentiation. Clin. Cancer Res. 1999, 5, 1465–1472.
104. Yu, H.N.; Zhang, L.C.; Yang, J.G.; Das, U.N.; Shen, S.R. Effect of laminin
tyrosine-isoleucine-glycine-serine-arginine peptide on the growth of human prostate cancer (pc-3) cells in
vitro. Eur. J. Pharmacol. 2009, 616, 251–255.
105. Liu, L.; Sun, L.; Zhang, H.; Li, Z.; Ning, X.; Shi, Y.; Guo, C.; Han, S.; Wu, K.; Fan, D. Hypoxia-mediated
up-regulation of mgr1-ag/37lrp in gastric cancers occurs via hypoxia-inducible-factor 1-dependent
mechanism and contributes to drug resistance. Int. J. Cancer 2009, 124, 1707–1715.
106. Pesapane, A.; Ragno, P.; Selleri, C.; Montuori, N. Recent advances in the function of the 67 kda laminin
receptor and its targeting for personalized therapy in cancer. Curr. Pharm. Des. 2017, 23, 4745–4757.
107. Pesapane, A.; Di Giovanni, C.; Rossi, F.W.; Alfano, D.; Formisano, L.; Ragno, P.; Selleri, C.; Montuori, N.;
Lavecchia, A. Discovery of new small molecules inhibiting 67 kda laminin receptor interaction with
laminin and cancer cell invasion. Oncotarget 2015, 6, 18116–18133.
108. Li, Y.; Li, D.; Chen, J.; Wang, S. A polysaccharide from pinellia ternata inhibits cell proliferation and
metastasis in human cholangiocarcinoma cells by targeting of cdc42 and 67kda laminin receptor (lr). Int. J.
Biol. Macromol. 2016, 93, 520–525.
109. Fujimura, Y.; Sumida, M.; Sugihara, K.; Tsukamoto, S.; Yamada, K.; Tachibana, H. Green tea polyphenol
egcg sensing motif on the 67-kda laminin receptor. PLoS ONE 2012, 7, e37942.
110. Fujimura, Y.; Yamada, K.; Tachibana, H. A lipid raft-associated 67kda laminin receptor mediates
suppressive effect of epigallocatechin-3-o-gallate on fcepsilonri expression. Biochem. Biophys. Res. Commun.
2005, 336, 674–681.
111. Simons, K.; Ikonen, E. Functional rafts in cell membranes. Nature 1997, 387, 569–572.
112. Xu, L.; Auzins, A.; Sun, X.; Xu, Y.; Harnischfeger, F.; Lu, Y.; Li, Z.; Chen, Y.H.; Zheng, W.; Liu, W. The
synaptic recruitment of lipid rafts is dependent on cd19-pi3k module and cytoskeleton remodeling
molecules. J. Leukoc. Biol. 2015, 98, 223–234.
113. Varshney, P.; Yadav, V.; Saini, N. Lipid rafts in immune signalling: Current progress and future
perspective. Immunology 2016, 149, 13–24.
114. Simons, K.; Toomre, D. Lipid rafts and signal transduction. Nat. Rev. Mol. Cell. Biol. 2000, 1, 31–39.
115. Pike, L.J. Growth factor receptors, lipid rafts and caveolae: An evolving story. Biochim. Biophys. Acta 2005,
1746, 260–273.
116. Diluvio, G.; Del Gaudio, F.; Giuli, M.V.; Franciosa, G.; Giuliani, E.; Palermo, R.; Besharat, Z.M.; Pignataro,
M.G.; Vacca, A.; d’Amati, G.; et al. Notch3 inactivation increases triple negative breast cancer sensitivity to
gefitinib by promoting egfr tyrosine dephosphorylation and its intracellular arrest. Oncogenesis 2018, 7, 42.
117. Pike, L.J.; Han, X.; Gross, R.W. Epidermal growth factor receptors are localized to lipid rafts that contain a
balance of inner and outer leaflet lipids: A shotgun lipidomics study. J. Biol. Chem. 2005, 280, 26796–26804.
118. Masuda, M.; Wakasaki, T.; Toh, S.; Shimizu, M.; Adachi, S. Chemoprevention of head and neck cancer by
green tea extract: Egcg-the role of egfr signaling and “lipid raft”. J. Oncol. 2011, 2011, 540148.
119. Guo, T.; Xu, L.; Che, X.; Zhang, S.; Li, C.; Wang, J.; Gong, J.; Ma, R.; Fan, Y.; Hou, K.; et al. Formation of the
igf1r/cav1/src tri-complex antagonizes trail-induced apoptosis in gastric cancer cells. Cell. Biol. Int. 2017, 41,
749–760.
120. Alawin, O.A.; Ahmed, R.A.; Ibrahim, B.A.; Briski, K.P.; Sylvester, P.W. Antiproliferative effects of
gamma-tocotrienol are associated with lipid raft disruption in her2-positive human breast cancer cells. J.
Nutr. Biochem. 2016, 27, 266–277.
121. Sur, S.; Pal, D.; Roy, R.; Barua, A.; Roy, A.; Saha, P.; Panda, C.K. Tea polyphenols egcg and tf restrict
tongue and liver carcinogenesis simultaneously induced by n-nitrosodiethylamine in mice. Toxicol. Appl.
Pharmacol. 2016, 300, 34–46.
122. Ma, Y.C.; Li, C.; Gao, F.; Xu, Y.; Jiang, Z.B.; Liu, J.X.; Jin, L.Y. Epigallocatechin gallate inhibits the growth of
human lung cancer by directly targeting the egfr signaling pathway. Oncol. Rep. 2014, 31, 1343–1349.
Nutrients 2018, 10, 1936 19 of 24
123. Filippi, A.; Picot, T.; Aanei, C.M.; Nagy, P.; Szollosi, J.; Campos, L.; Ganea, C.; Mocanu, M.M.
Epigallocatechin-3-o-gallate alleviates the malignant phenotype in a-431 epidermoid and sk-br-3 breast
cancer cell lines. Int. J. Food Sci. Nutr. 2018, 69, 584–597.
124. Pike, L.J. Rafts defined: A report on the keystone symposium on lipid rafts and cell function. J. Lipid. Res.
2006, 47, 1597–1598.
125. Kim, J.Y.; Wang, L.; Lee, J.; Ou, J.J. Hepatitis c virus induces the localization of lipid rafts to
autophagosomes for its rna replication. J. Virol. 2017, 91, doi:10.1128/JVI.00541-17.
126. Rosenberger, C.M.; Brumell, J.H.; Finlay, B.B. Microbial pathogenesis: Lipid rafts as pathogen portals.
Curr. Biol. 2000, 10, R823–R825.
127. Guimaraes, A.J.; de Cerqueira, M.D.; Zamith-Miranda, D.; Lopez, P.H.; Rodrigues, M.L.; Pontes, B.; Viana,
N.B.; DeLeon-Rodriguez, C.M.; Rossi, D.C.P.; Casadevall, A.; et al. Host membrane glycosphingolipids
and lipid microdomains facilitate histoplasma capsulatum internalization by macrophages. Cell. Microbiol.
2018, e12976, doi:10.1111/cmi.12976.
128. Smart, E.J.; Graf, G.A.; McNiven, M.A.; Sessa, W.C.; Engelman, J.A.; Scherer, P.E.; Okamoto, T.; Lisanti,
M.P. Caveolins, liquid-ordered domains, and signal transduction. Mol. Cell. Biol. 1999, 19, 7289–7304.
129. Hwangbo, C.; Tae, N.; Lee, S.; Kim, O.; Park, O.K.; Kim, J.; Kwon, S.H.; Lee, J.H. Syntenin regulates
tgf-beta1-induced smad activation and the epithelial-to-mesenchymal transition by inhibiting
caveolin-mediated tgf-beta type i receptor internalization. Oncogene 2016, 35, 389–401.
130. Laurenzana, A.; Fibbi, G.; Chilla, A.; Margheri, G.; Del Rosso, T.; Rovida, E.; Del Rosso, M.; Margheri, F.
Lipid rafts: Integrated platforms for vascular organization offering therapeutic opportunities. Cell. Mol.
Life Sci. 2015, 72, 1537–1557.
131. Mollinedo, F.; Gajate, C. Lipid rafts as major platforms for signaling regulation in cancer. Adv. Biol. Regul.
2015, 57, 130–146.
132. Tai, Y.T.; Podar, K.; Catley, L.; Tseng, Y.H.; Akiyama, M.; Shringarpure, R.; Burger, R.; Hideshima, T.;
Chauhan, D.; Mitsiades, N.; et al. Insulin-like growth factor-1 induces adhesion and migration in human
multiple myeloma cells via activation of beta1-integrin and phosphatidylinositol 3’-kinase/akt signaling.
Cancer Res. 2003, 63, 5850–5858.
133. Raghu, H.; Sodadasu, P.K.; Malla, R.R.; Gondi, C.S.; Estes, N.; Rao, J.S. Localization of upar and mmp-9 in
lipid rafts is critical for migration, invasion and angiogenesis in human breast cancer cells. BMC Cancer
2010, 10, 647.
134. Lacour, S.; Hammann, A.; Grazide, S.; Lagadic-Gossmann, D.; Athias, A.; Sergent, O.; Laurent, G.;
Gambert, P.; Solary, E.; Dimanche-Boitrel, M.T. Cisplatin-induced cd95 redistribution into membrane lipid
rafts of ht29 human colon cancer cells. Cancer Res. 2004, 64, 3593–3598.
135. George, K.S.; Wu, S. Lipid raft: A floating island of death or survival. Toxicol. Appl. Pharmacol. 2012, 259,
311–319.
136. Alves, A.C.S.; Dias, R.A.; Kagami, L.P.; das Neves, G.M.; Torres, F.C.; Eifler-Lima, V.L.; Carvalho, I.; de
Miranda Silva, C.; Kawano, D.F. Beyond the “lock and key” paradigm: Targeting lipid rafts to induce the
selective apoptosis of cancer cells. Curr. Med. Chem. 2018, 25, 2082–2104.
137. Tsukamoto, S.; Hirotsu, K.; Kumazoe, M.; Goto, Y.; Sugihara, K.; Suda, T.; Tsurudome, Y.; Suzuki, T.;
Yamashita, S.; Kim, Y.; et al. Green tea polyphenol egcg induces lipid-raft clustering and apoptotic cell
death by activating protein kinase cdelta and acid sphingomyelinase through a 67 kda laminin receptor in
multiple myeloma cells. Biochem. J. 2012, 443, 525–534.
138. Mocanu, M.M.; Ganea, C.; Georgescu, L.; Varadi, T.; Shrestha, D.; Baran, I.; Katona, E.; Nagy, P.; Szollosi, J.
Epigallocatechin 3-o-gallate induces 67 kda laminin receptor-mediated cell death accompanied by
downregulation of erbb proteins and altered lipid raft clustering in mammary and epidermoid carcinoma
cells. J. Nat. Prod. 2014, 77, 250–257.
139. Huang, Y.; Kumazoe, M.; Bae, J.; Yamada, S.; Takai, M.; Hidaka, S.; Yamashita, S.; Kim, Y.; Won, Y.;
Murata, M.; et al. Green tea polyphenol epigallocatechin-o-gallate induces cell death by acid
sphingomyelinase activation in chronic myeloid leukemia cells. Oncol. Rep. 2015, 34, 1162–1168.
140. Yang, C.S.; Wang, H. Cancer preventive activities of tea catechins. Molecules 2016, 21, 1679.
141. Luo, K.W.; Lung, W.Y.; Chun, X.; Luo, X.L.; Huang, W.R. Egcg inhibited bladder cancer t24 and 5637 cell
proliferation and migration via pi3k/akt pathway. Oncotarget 2018, 9, 12261–12272.
Nutrients 2018, 10, 1936 20 of 24
142. Velavan, B.; Divya, T.; Sureshkumar, A.; Sudhandiran, G. Nano-chemotherapeutic efficacy of (-)
-epigallocatechin 3-gallate mediating apoptosis in a549cells: Involvement of reactive oxygen species
mediated nrf2/keap1signaling. Biochem. Biophys. Res. Commun. 2018, 503, 1723–1731.
143. Gu, J.J.; Qiao, K.S.; Sun, P.; Chen, P.; Li, Q. Study of egcg induced apoptosis in lung cancer cells by
inhibiting pi3k/akt signaling pathway. Eur. Rev. Med. Pharmacol. Sci. 2018, 22, 4557–4563.
144. Wang, Y.Q.; Lu, J.L.; Liang, Y.R.; Li, Q.S. Suppressive effects of egcg on cervical cancer. Molecules 2018, 23,
2334.
145. Britschgi, A.; Simon, H.U.; Tobler, A.; Fey, M.F.; Tschan, M.P. Epigallocatechin-3-gallate induces cell death
in acute myeloid leukaemia cells and supports all-trans retinoic acid-induced neutrophil differentiation
via death-associated protein kinase 2. Br. J. Haematol. 2010, 149, 55–64.
146. Shammas, M.A.; Neri, P.; Koley, H.; Batchu, R.B.; Bertheau, R.C.; Munshi, V.; Prabhala, R.; Fulciniti, M.;
Tai, Y.T.; Treon, S.P.; et al. Specific killing of multiple myeloma cells by (-)-epigallocatechin-3-gallate
extracted from green tea: Biologic activity and therapeutic implications. Blood 2006, 108, 2804–2810.
147. Kirschnek, S.; Paris, F.; Weller, M.; Grassme, H.; Ferlinz, K.; Riehle, A.; Fuks, Z.; Kolesnick, R.; Gulbins, E.
Cd95-mediated apoptosis in vivo involves acid sphingomyelinase. J. Biol. Chem. 2000, 275, 27316–27323.
148. Garcia-Barros, M.; Paris, F.; Cordon-Cardo, C.; Lyden, D.; Rafii, S.; Haimovitz-Friedman, A.; Fuks, Z.;
Kolesnick, R. Tumor response to radiotherapy regulated by endothelial cell apoptosis. Science 2003, 300,
1155–1159.
149. Zhang, Y.; Mattjus, P.; Schmid, P.C.; Dong, Z.; Zhong, S.; Ma, W.Y.; Brown, R.E.; Bode, A.M.; Schmid, H.H.
Involvement of the acid sphingomyelinase pathway in uva-induced apoptosis. J. Biol. Chem. 2001, 276,
11775–11782.
150. Goni, F.M.; Alonso, A. Sphingomyelinases: Enzymology and membrane activity. FEBS Lett. 2002, 531, 38–
46.
151. Morita, Y.; Perez, G.I.; Paris, F.; Miranda, S.R.; Ehleiter, D.; Haimovitz-Friedman, A.; Fuks, Z.; Xie, Z.;
Reed, J.C.; Schuchman, E.H.; et al. Oocyte apoptosis is suppressed by disruption of the acid
sphingomyelinase gene or by sphingosine-1-phosphate therapy. Nat. Med. 2000, 6, 1109–1114.
152. Gulbins, E. Regulation of death receptor signaling and apoptosis by ceramide. Pharmacol. Res. 2003, 47,
393–399.
153. Hueber, A.O.; Bernard, A.M.; Herincs, Z.; Couzinet, A.; He, H.T. An essential role for membrane rafts in
the initiation of fas/cd95-triggered cell death in mouse thymocytes. EMBO Rep. 2002, 3, 190–196.
154. Kischkel, F.C.; Hellbardt, S.; Behrmann, I.; Germer, M.; Pawlita, M.; Krammer, P.H.; Peter, M.E.
Cytotoxicity-dependent apo-1 (fas/cd95)-associated proteins form a death-inducing signaling complex
(disc) with the receptor. EMBO J. 1995, 14, 5579–5588.
155. Grassme, H.; Cremesti, A.; Kolesnick, R.; Gulbins, E. Ceramide-mediated clustering is required for
cd95-disc formation. Oncogene 2003, 22, 5457–5470.
156. Gajate, C.; Mollinedo, F. The antitumor ether lipid et-18-och(3) induces apoptosis through translocation
and capping of fas/cd95 into membrane rafts in human leukemic cells. Blood 2001, 98, 3860–3863.
157. London, E. Ceramide selectively displaces cholesterol from ordered lipid domains (rafts): Implications for
lipid raft structure and function. J. Biol. Chem. 2004, 279, 9997–10004.
158. Cremesti, A.; Paris, F.; Grassme, H.; Holler, N.; Tschopp, J.; Fuks, Z.; Gulbins, E.; Kolesnick, R. Ceramide
enables fas to cap and kill. J. Biol. Chem. 2001, 276, 23954–23961.
159. Fanzo, J.C.; Lynch, M.P.; Phee, H.; Hyer, M.; Cremesti, A.; Grassme, H.; Norris, J.S.; Coggeshall, K.M.;
Rueda, B.R.; Pernis, A.B.; et al. Cd95 rapidly clusters in cells of diverse origins. Cancer Biol. Ther. 2003, 2,
392–395.
160. Wu, L.Y.; De Luca, T.; Watanabe, T.; Morre, D.M.; Morre, D.J. Metabolite modulation of hela cell response
to enox2 inhibitors egcg and phenoxodiol. Biochim. Biophys. Acta 2011, 1810, 784–789.
161. Kim, M.H.; Chung, J. Synergistic cell death by egcg and ibuprofen in du-145 prostate cancer cell line.
Anticancer Res. 2007, 27, 3947–3956.
162. Tan, X.; Zhang, Y.; Jiang, B.; Zhou, D. Changes in ceramide levels upon catechins-induced apoptosis in
lovo cells. Life Sci. 2002, 70, 2023–2029.
163. Zhou, L.; Yang, F.; Li, G.; Huang, J.; Liu, Y.; Zhang, Q.; Tang, Q.; Hu, C.; Zhang, R. Coptisine induces
apoptosis in human hepatoma cells through activating 67-kda laminin receptor/cgmp signaling. Front.
Pharmacol 2018, 9, 517.
Nutrients 2018, 10, 1936 21 of 24
164. Kumazoe, M.; Kim, Y.; Bae, J.; Takai, M.; Murata, M.; Suemasu, Y.; Sugihara, K.; Yamashita, S.; Tsukamoto,
S.; Huang, Y.; et al. Phosphodiesterase 5 inhibitor acts as a potent agent sensitizing acute myeloid
leukemia cells to 67-kda laminin receptor-dependent apoptosis. FEBS Lett. 2013, 587, 3052–3057.
165. Kumazoe, M.; Tsukamoto, S.; Lesnick, C.; Kay, N.E.; Yamada, K.; Shanafelt, T.D.; Tachibana, H.
Vardenafil, a clinically available phosphodiesterase inhibitor, potentiates the killing effect of egcg on cll
cells. Br. J. Haematol. 2015, 168, 610–613.
166. Casaletto, J.B.; McClatchey, A.I. Spatial regulation of receptor tyrosine kinases in development and cancer.
Nat. Rev. Cancer 2012, 12, 387–400.
167. Brizuela, L.; Dayon, A.; Doumerc, N.; Ader, I.; Golzio, M.; Izard, J.C.; Hara, Y.; Malavaud, B.; Cuvillier, O.
The sphingosine kinase-1 survival pathway is a molecular target for the tumor-suppressive tea and wine
polyphenols in prostate cancer. Faseb J. 2010, 24, 3882–3894.
168. Olivera, A.; Spiegel, S. Sphingosine-1-phosphate as second messenger in cell proliferation induced by pdgf
and fcs mitogens. Nature 1993, 365, 557–560.
169. Cuvillier, O.; Pirianov, G.; Kleuser, B.; Vanek, P.G.; Coso, O.A.; Gutkind, S.; Spiegel, S. Suppression of
ceramide-mediated programmed cell death by sphingosine-1-phosphate. Nature 1996, 381, 800–803.
170. Tsukamoto, S.; Kumazoe, M.; Huang, Y.; Lesnick, C.; Kay, N.E.; Shanafelt, T.D.; Tachibana, H. Sphk1
inhibitor potentiates the anti-cancer effect of egcg on leukaemia cells. Br. J. Haematol. 2017, 178, 155–158.
171. Chakrabarty, S.; Ganguli, A.; Das, A.; Nag, D.; Chakrabarti, G. Epigallocatechin-3-gallate shows
anti-proliferative activity in hela cells targeting tubulin-microtubule equilibrium. Chem. Biol. Interact. 2015,
242, 380–389.
172. Shenouda, N.S.; Zhou, C.; Browning, J.D.; Ansell, P.J.; Sakla, M.S.; Lubahn, D.B.; Macdonald, R.S.
Phytoestrogens in common herbs regulate prostate cancer cell growth in vitro. Nutr. Cancer 2004, 49, 200–
208.
173. Umeda, D.; Yano, S.; Yamada, K.; Tachibana, H. Involvement of 67-kda laminin receptor-mediated myosin
phosphatase activation in antiproliferative effect of epigallocatechin-3-O-gallate at a physiological
concentration on caco-2 colon cancer cells. Biochem. Biophys. Res. Commun. 2008, 371, 172–176.
174. D’Avino, P.P.; Giansanti, M.G.; Petronczki, M. Cytokinesis in animal cells. Cold Spring Harb. Perspect. Biol.
2015, 7, a015834.
175. Wang, Y.L. The mechanism of cortical ingression during early cytokinesis: Thinking beyond the
contractile ring hypothesis. Trends Cell. Biol. 2005, 15, 581–588.
176. Matsumura, F. Regulation of myosin ii during cytokinesis in higher eukaryotes. Trends Cell. Biol. 2005, 15,
371–377.
177. Moussavi, R.S.; Kelley, C.A.; Adelstein, R.S. Phosphorylation of vertebrate nonmuscle and smooth muscle
myosin heavy chains and light chains. Mol. Cell. Biochem. 1993, 127–128, 219–227.
178. Scholey, J.M.; Taylor, K.A.; Kendrick-Jones, J. Regulation of non-muscle myosin assembly by
calmodulin-dependent light chain kinase. Nature 1980, 287, 233–235.
179. Ikebe, M.; Koretz, J.; Hartshorne, D.J. Effects of phosphorylation of light chain residues threonine 18 and
serine 19 on the properties and conformation of smooth muscle myosin. J. Biol. Chem. 1988, 263, 6432–6437.
180. Kawano, Y.; Fukata, Y.; Oshiro, N.; Amano, M.; Nakamura, T.; Ito, M.; Matsumura, F.; Inagaki, M.;
Kaibuchi, K. Phosphorylation of myosin-binding subunit (mbs) of myosin phosphatase by rho-kinase in
vivo. J. Cell. Biol. 1999, 147, 1023–1038.
181. Negrutskii, B.S.; El’skaya, A.V. Eukaryotic translation elongation factor 1 alpha: Structure, expression,
functions, and possible role in aminoacyl-trna channeling. Prog. Nucleic Acid Res. Mol. Biol. 1998, 60, 47–78.
182. Gangwani, L.; Mikrut, M.; Galcheva-Gargova, Z.; Davis, R.J. Interaction of zpr1 with translation
elongation factor-1alpha in proliferating cells. J. Cell. Biol. 1998, 143, 1471–1484.
183. Lamberti, A.; Caraglia, M.; Longo, O.; Marra, M.; Abbruzzese, A.; Arcari, P. The translation elongation
factor 1a in tumorigenesis, signal transduction and apoptosis: Review article. Amino Acids 2004, 26, 443–
448.
184. Izawa, T.; Fukata, Y.; Kimura, T.; Iwamatsu, A.; Dohi, K.; Kaibuchi, K. Elongation factor-1 alpha is a novel
substrate of rho-associated kinase. Biochem. Biophys. Res. Commun. 2000, 278, 72–78.
185. Peterson, R.T.; Desai, B.N.; Hardwick, J.S.; Schreiber, S.L. Protein phosphatase 2a interacts with the 70-kda
s6 kinase and is activated by inhibition of fkbp12-rapamycinassociated protein. Proc. Natl. Acad. Sci. USA
1999, 96, 4438–4442.
Nutrients 2018, 10, 1936 22 of 24
186. Zhang, Q.; Claret, F.X. Phosphatases: The new brakes for cancer development? Enzyme Res. 2012, 2012,
659649.
187. Kiss, A.; Becsi, B.; Kolozsvari, B.; Komaromi, I.; Kover, K.E.; Erdodi, F. Epigallocatechin-3-gallate and
penta-o-galloyl-beta-d-glucose inhibit protein phosphatase-1. FEBS J. 2013, 280, 612–626.
188. Kitano, K.; Nam, K.Y.; Kimura, S.; Fujiki, H.; Imanishi, Y. Sealing effects of (-)-epigallocatechin gallate on
protein kinase c and protein phosphatase 2a. Biophys. Chem. 1997, 65, 157–164.
189. Janssens, V.; Goris, J.; Van Hoof, C. Pp2a: The expected tumor suppressor. Curr. Opin. Genet. Dev. 2005, 15,
34–41.
190. Stamenkovic, I.; Yu, Q. Merlin, a “magic” linker between extracellular cues and intracellular signaling
pathways that regulate cell motility, proliferation, and survival. Curr. Protein Pept. Sci. 2010, 11, 471–484.
191. Horiguchi, A.; Zheng, R.; Shen, R.; Nanus, D.M. Inactivation of the nf2 tumor suppressor protein merlin in
du145 prostate cancer cells. Prostate 2008, 68, 975–984.
192. Malhotra, A.; Shibata, Y.; Hall, I.M.; Dutta, A. Chromosomal structural variations during progression of a
prostate epithelial cell line to a malignant metastatic state inactivate the nf2, nipsnap1, ugt2b17, and lpin2
genes. Cancer Biol. Ther. 2013, 14, 840–852.
193. Petrilli, A.M.; Fernandez-Valle, C. Role of merlin/nf2 inactivation in tumor biology. Oncogene 2016, 35,
537–548.
194. Ambros, V. Micrornas: Tiny regulators with great potential. Cell 2001, 107, 823–826.
195. Johnson, S.M.; Grosshans, H.; Shingara, J.; Byrom, M.; Jarvis, R.; Cheng, A.; Labourier, E.; Reinert, K.L.;
Brown, D.; Slack, F.J. Ras is regulated by the let-7 microrna family. Cell 2005, 120, 635–647.
196. Schultz, J.; Lorenz, P.; Gross, G.; Ibrahim, S.; Kunz, M. Microrna let-7b targets important cell cycle
molecules in malignant melanoma cells and interferes with anchorage-independent growth. Cell Res. 2008,
18, 549–557.
197. Zedan, A.H.; Hansen, T.F.; Assenholt, J.; Pleckaitis, M.; Madsen, J.S.; Osther, P.J.S. Microrna expression in
tumour tissue and plasma in patients with newly diagnosed metastatic prostate cancer. Tumour. Biol. 2018,
40, 1010428318775864.
198. Zhou, H.; Chen, J.X.; Yang, C.S.; Yang, M.Q.; Deng, Y.; Wang, H. Gene regulation mediated by micrornas
in response to green tea polyphenol egcg in mouse lung cancer. BMC Genom. 2014, 15 (Suppl. 11), S3.
199. Pan, X.; Zhao, B.; Song, Z.; Han, S.; Wang, M. Estrogen receptor-alpha36 is involved in
epigallocatechin-3-gallate induced growth inhibition of er-negative breast cancer stem/progenitor cells. J.
Pharmacol. Sci. 2016, 130, 85–93.
200. Jiang, P.; Xu, C.; Chen, L.; Chen, A.; Wu, X.; Zhou, M.; Haq, I.U.; Mariyam, Z.; Feng, Q. Egcg inhibits
csc-like properties through targeting mir-485/cd44 axis in a549-cisplatin resistant cells. Mol. Carcinog. 2018,
57, 1835–1844.
201. Nishimura, N.; Hartomo, T.B.; Pham, T.V.; Lee, M.J.; Yamamoto, T.; Morikawa, S.; Hasegawa, D.; Takeda,
H.; Kawasaki, K.; Kosaka, Y.; et al. Epigallocatechin gallate inhibits sphere formation of neuroblastoma
be(2)-c cells. Environ. Health Prev. Med. 2012, 17, 246–251.
202. Kumazoe, M.; Takai, M.; Hiroi, S.; Takeuchi, C.; Yamanouchi, M.; Nojiri, T.; Onda, H.; Bae, J.; Huang, Y.;
Takamatsu, K.; et al. Pde3 inhibitor and egcg combination treatment suppress cancer stem cell properties
in pancreatic ductal adenocarcinoma. Sci. Rep. 2017, 7, 1917.
203. Kumazoe, M.; Takai, M.; Bae, J.; Hiroi, S.; Huang, Y.; Takamatsu, K.; Won, Y.; Yamashita, M.; Hidaka, S.;
Yamashita, S.; et al. Foxo3 is essential for cd44 expression in pancreatic cancer cells. Oncogene 2017, 36,
2643–2654.
204. Eddy, S.F.; Kane, S.E.; Sonenshein, G.E. Trastuzumab-resistant her2-driven breast cancer cells are sensitive
to epigallocatechin-3 gallate. Cancer Res. 2007, 67, 9018–9023.
205. Farabegoli, F.; Govoni, M.; Ciavarella, C.; Orlandi, M.; Papi, A. A rxr ligand
6-oh-11-o-hydroxyphenanthrene with antitumour properties enhances (-)-epigallocatechin-3-gallate
activity in three human breast carcinoma cell lines. Biomed. Res. Int. 2014, 2014, 853086.
206. Lugnier, C. Cyclic nucleotide phosphodiesterase (pde) superfamily: A new target for the development of
specific therapeutic agents. Pharmacol. Ther. 2006, 109, 366–398.
207. Lu, K.P.; Zhou, X.Z. The prolyl isomerase pin1: A pivotal new twist in phosphorylation signalling and
disease. Nat. Rev. Mol. Cell Biol. 2007, 8, 904–916.
208. Dominguez-Sola, D.; Dalla-Favera, R. Pinning down the c-myc oncoprotein. Nat. Cell Biol. 2004, 6, 288–289.
209. Sears, R.C. The life cycle of c-myc: From synthesis to degradation. Cell Cycle 2004, 3, 1133–1137.
Nutrients 2018, 10, 1936 23 of 24
210. Ryo, A.; Nakamura, M.; Wulf, G.; Liou, Y.C.; Lu, K.P. Pin1 regulates turnover and subcellular localization
of beta-catenin by inhibiting its interaction with apc. Nat. Cell Biol. 2001, 3, 793–801.
211. Bao, L.; Kimzey, A.; Sauter, G.; Sowadski, J.M.; Lu, K.P.; Wang, D.G. Prevalent overexpression of prolyl
isomerase pin1 in human cancers. Am. J. Pathol. 2004, 164, 1727–1737.
212. Ayala, G.; Wang, D.; Wulf, G.; Frolov, A.; Li, R.; Sowadski, J.; Wheeler, T.M.; Lu, K.P.; Bao, L. The prolyl
isomerase pin1 is a novel prognostic marker in human prostate cancer. Cancer Res. 2003, 63, 6244–6251.
213. Moore, J.D.; Potter, A. Pin1 inhibitors: Pitfalls, progress and cellular pharmacology. Bioorg. Med. Chem. Lett.
2013, 23, 4283–4291.
214. Hidaka, M.; Kosaka, K.; Tsushima, S.; Uchida, C.; Takahashi, K.; Takahashi, N.; Tsubuki, M.; Hara, Y.;
Uchida, T. Food polyphenols targeting peptidyl prolyl cis/trans isomerase pin1. Biochem. Biophys. Res.
Commun. 2018, 499, 681–687.
215. Xi, L.; Wang, Y.; He, Q.; Zhang, Q.; Du, L. Interaction between pin1 and its natural product inhibitor
epigallocatechin-3-gallate by spectroscopy and molecular dynamics simulations. Spectrochim. Acta A Mol.
Biomol. Spectrosc 2016, 169, 134–143.
216. Katsuno, Y.; Lamouille, S.; Derynck, R. Tgf-beta signaling and epithelial-mesenchymal transition in cancer
progression. Curr. Opin. Oncol. 2013, 25, 76–84.
217. Sun, L.; Diamond, M.E.; Ottaviano, A.J.; Joseph, M.J.; Ananthanarayan, V.; Munshi, H.G. Transforming
growth factor-beta 1 promotes matrix metalloproteinase-9-mediated oral cancer invasion through snail
expression. Mol. Cancer Res. 2008, 6, 10–20.
218. Joseph, M.J.; Dangi-Garimella, S.; Shields, M.A.; Diamond, M.E.; Sun, L.; Koblinski, J.E.; Munshi, H.G. Slug
is a downstream mediator of transforming growth factor-beta1-induced matrix metalloproteinase-9
expression and invasion of oral cancer cells. J. Cell. Biochem. 2009, 108, 726–736.
219. Sinpitaksakul, S.N.; Pimkhaokham, A.; Sanchavanakit, N.; Pavasant, P. Tgf-beta1 induced mmp-9
expression in hnscc cell lines via smad/mlck pathway. Biochem. Biophys. Res. Commun. 2008, 371, 713–718.
220. Chowdhury, A.; Nandy, S.K.; Sarkar, J.; Chakraborti, T.; Chakraborti, S. Inhibition of pro-/active mmp-2
by green tea catechins and prediction of their interaction by molecular docking studies. Mol. Cell. Biochem.
2017, 427, 111–122.
221. Sarkar, J.; Nandy, S.K.; Chowdhury, A.; Chakraborti, T.; Chakraborti, S. Inhibition of mmp-9 by green tea
catechins and prediction of their interaction by molecular docking analysis. Biomed. Pharmacother. 2016, 84,
340–347.
222. Schramm, L. Going green: The role of the green tea component egcg in chemoprevention. J. Carcinog.
Mutagen. 2013, 4, 1000142.
223. Riley, P.A. Epimutation and cancer: Carcinogenesis viewed as error-prone inheritance of epigenetic
information. J. Oncol. 2018, 2018, 2645095.
224. Huang, Z.; Huang, Q.; Ji, L.; Wang, Y.; Qi, X.; Liu, L.; Liu, Z.; Lu, L. Epigenetic regulation of active Chinese
herbal components for cancer prevention and treatment: A follow-up review. Pharmacol. Res. 2016, 114, 1–
12.
225. Fang, M.Z.; Wang, Y.; Ai, N.; Hou, Z.; Sun, Y.; Lu, H.; Welsh, W.; Yang, C.S. Tea polyphenol
(-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes
in cancer cell lines. Cancer Res. 2003, 63, 7563–7570.
226. Pal, D.; Sur, S.; Roy, R.; Mandal, S.; Kumar Panda, C. Epigallocatechin gallate in combination with eugenol
or amarogentin shows synergistic chemotherapeutic potential in cervical cancer cell line. J. Cell. Physiol.
2018, 234, 825–836.
227. Oya, Y.; Mondal, A.; Rawangkan, A.; Umsumarng, S.; Iida, K.; Watanabe, T.; Kanno, M.; Suzuki, K.; Li, Z.;
Kagechika, H.; et al. Down-regulation of histone deacetylase 4, -5 and -6 as a mechanism of synergistic
enhancement of apoptosis in human lung cancer cells treated with the combination of a synthetic retinoid,
am80 and green tea catechin. J. Nutr. Biochem. 2017, 42, 7–16.
228. Zhang, Y.; Wang, X.; Han, L.; Zhou, Y.; Sun, S. Green tea polyphenol egcg reverse cisplatin resistance of
a549/ddp cell line through candidate genes demethylation. Biomed. Pharmacother. 2015, 69, 285–290.
229. Jin, H.; Chen, J.X.; Wang, H.; Lu, G.; Liu, A.; Li, G.; Tu, S.; Lin, Y.; Yang, C.S. Nnk-induced DNA
methyltransferase 1 in lung tumorigenesis in a/j mice and inhibitory effects of
(-)-epigallocatechin-3-gallate. Nutr. Cancer 2015, 67, 167–176.
230. Liu, L.; Zuo, J.; Wang, G. Epigallocatechin-3-gallate suppresses cell proliferation and promotes apoptosis
in ec9706 and eca109 esophageal carcinoma cells. Oncol. Lett. 2017, 14, 4391–4395.
Nutrients 2018, 10, 1936 24 of 24
231. Le, C.T.; Leenders, W.P.J.; Molenaar, R.J.; van Noorden, C.J.F. Effects of the green tea polyphenol
epigallocatechin-3-gallate on glioma: A critical evaluation of the literature. Nutr. Cancer 2018, 70, 317–333.
232. Li, W.G.; Li, Q.H.; Tan, Z. Epigallocatechin gallate induces telomere fragmentation in hela and 293 but not
in mrc-5 cells. Life Sci. 2005, 76, 1735–1746.
233. Zhang, W.; Yang, P.; Gao, F.; Yang, J.; Yao, K. Effects of epigallocatechin gallate on the proliferation and
apoptosis of the nasopharyngeal carcinoma cell line cne2. Exp. Ther. Med. 2014, 8, 1783–1788.
234. Wang, X.; Hao, M.W.; Dong, K.; Lin, F.; Ren, J.H.; Zhang, H.Z. Apoptosis induction effects of egcg in
laryngeal squamous cell carcinoma cells through telomerase repression. Arch. Pharm. Res. 2009, 32, 1263–
1269.
235. Sadava, D.; Whitlock, E.; Kane, S.E. The green tea polyphenol, epigallocatechin-3-gallate inhibits
telomerase and induces apoptosis in drug-resistant lung cancer cells. Biochem. Biophys. Res. Commun. 2007,
360, 233–237.
236. Kuzuhara, T.; Sei, Y.; Yamaguchi, K.; Suganuma, M.; Fujiki, H. DNA and rna as new binding targets of
green tea catechins. J. Biol. Chem. 2006, 281, 17446–17456.
237. Kuzuhara, T.; Tanabe, A.; Sei, Y.; Yamaguchi, K.; Suganuma, M.; Fujiki, H. Synergistic effects of multiple
treatments, and both DNA and rna direct bindings on, green tea catechins. Mol. Carcinog. 2007, 46, 640–
645.
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... Catechins display many anti-carcinogenic and anti-mutagenic promising protective effects in cancer, including breast, esophagus, prostate, stomach, small intestine, colon, liver, and lung [2]. A number of in vitro, in vivo, and clinical studies have established that catechins produced their anticancer effects by way of the modification of several signaling routes, most of which are provoked by the interaction between catechins and an array of membrane proteins, intracellular molecules, membrane microdomains, and the plasma membrane itself [3]. ...
... Catechins altered the lipid order in ht29 colon cancer cells causing inhibition of epidermal growth factor receptor [8], and affected the lipid rafts blocking activation of the c-met receptor in prostate cancer cells [9]. In addition, there is evidence that one of the principal processes whereby catechins exert their anticancer action is the induction of lipid raft mediated apoptosis [3]. Catechins may exert their effects at the membrane level or inside the cell, but in both cases, during their therapeutic route, the interaction with the membrane becomes obligatory, hence this interaction should be studied in order to get insight into their mechanism of action. ...
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3,4,5-Trimethoxybenzoate of catechin (TMBC) is a semisynthetic catechin which shows strong antiproliferative activity against malignant melanoma cells. The amphiphilic nature of the molecule suggests that the membrane could be a potential site of action, hence the study of its interaction with lipid bilayers is mandatory in order to gain information on the effect of the catechin on the membrane properties and dynamics. Anionic phospholipids, though being minor components of the membrane, possess singular physical and biochemical properties that make them physiologically essential. Utilizing phosphatidylserine biomimetic membranes, we study the interaction between the catechin and anionic bilayers, bringing together a variety of experimental techniques and molecular dynamics simulation. The experimental data suggest that the molecule is embedded into the phosphatidylserine bilayers, where it perturbs the thermotropic gel to liquid crystalline phase transition. In the gel phase, the catechin promotes the formation of interdigitation, and in the liquid crystalline phase, it decreases the bilayer thickness and increases the hydrogen bonding pattern of the interfacial region of the bilayer. The simulation data agree with the experimental ones and indicate that the molecule is located in the interior of the anionic bilayer as monomer and small clusters reaching the carbonyl region of the phospholipid, where it also disturbs the intermolecular hydrogen bonding between neighboring lipids. Our observations suggest that the catechin incorporates well into phosphatidylserine bilayers, where it produces structural changes that could affect the functioning of the membrane.
... Several antitumor mechanisms were reported, including apoptosis activation, initiation of cellular growth block, modification of cellular life regulatory proteins, induction of killer caspases, and NF-kB pathway inhibition [89,90]. EGCG attenuated molecular pathways implicated in tumor advance [92] and in modulation of the immune system in murine cancers [93][94][95][96]. Rawangkan et al. [97] hypothesized that EGCG inhibited PD-L1, a checkpoint molecule, increasing anticancer immune response [97]. ...
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Cancer is one of the leading causes of death globally. Anticancer drugs aim to block tumor growth by killing cancerous cells in order to prevent tumor progression and metastasis. Efficient anticancer drugs should also minimize general toxicity towards organs and healthy cells. Tumor growth can also be successfully restrained by targeting and modulating immune response. Cancer immunotherapy is assuming a growing relevance in the fight against cancer and has recently aroused much interest for its wider safety and the capability to complement conventional chemotherapeutic approaches. Natural products are a traditional source of molecules with relevant potential in the pharmacological field. The huge structural diversity of metabolites with low molecular weight (small molecules) from terrestrial and marine organisms has provided lead compounds for the discovery of many modern anticancer drugs. Many natural products combine chemo-protective and immunomod-ulant activity, thus offering the potential to be used alone or in association with conventional cancer therapy. In this review, we report the natural products known to possess antitumor properties by interaction with immune system, as well as discuss the possible immunomodulatory mechanisms of these molecules.
... Although numerous health benefits of EGCG have been recognized, the cellular and molecular mechanisms behind them are not yet fully clarified, probably because the action of EGCG impacts multiple cellular pathways, thus simultaneously affecting several processes [21]. Nonetheless, EGCG has emerged as a chemopreventive, and potentially antioncogenic product, for the same reason of being capable to target different signaling cascades. ...
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Autophagy is an evolutionarily conserved process for the degradation of redundant or damaged cellular material by means of a lysosome-dependent mechanism, contributing to cell homeostasis and survival. Autophagy plays a multifaceted and context-dependent role in cancer initiation, maintenance, and progression; it has a tumor suppressive role in the absence of disease and is upregulated in cancer cells to meet their elevated metabolic demands. Autophagy represents a promising but challenging target in cancer treatment. Green tea is a widely used beverage with healthy effects on several diseases, including cancer. The bioactive compounds of green tea are mainly catechins, and epigallocatechin-gallate (EGCG) is the most abundant and biologically active among them. In this review, evidence of autophagy modulation and anti-cancer effects induced by EGCG treatment in experimental cancer models is presented. Reviewed articles reveal that EGCG promotes cytotoxic autophagy often through the inactivation of PI3K/Akt/mTOR pathway, resulting in apoptosis induction. EGCG pro-oxidant activity has been postulated to be responsible for its anti-cancer effects. In combination therapy with a chemotherapy drug, EGCG inhibits cell growth and the drug-induced pro-survival autophagy. The selected studies rightly claim EGCG as a valuable agent in cancer chemoprevention.
... Enzymes of histone deacetylase (HDACs) and DNA methyltransferase (DNMTs) have the main role in suppressing transcriptional genes and histone acetyltransferase (HAT) enzymes positively modulate gene expression regulation. Several studies have indicated the EGCG effect in epigenetic management acting on HATs, HDACs, and DNMTs expression/activity in different types of cancer [30,31]. Sheng et al. proved that EGCG (20 μM EGCG) reduced DNMT expression in the treated MDA-MB-231 and MCF-7 breast cancer cells [32]. ...
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Anaplastic thyroid cancer cases with poor prognosis are associated with epigenetic modifications such as abnormal DNA methylation. Epigallocathesin-3-gallate (EGCG) is a polyphenol compound of green tea that is still under investigation on its role in cancer prevention. EGCG is known as an epigenetic diet in DNA methyltransferase inhibitor. The cytotoxic effects of Dabrafenib, EGCG, and dabrafenib in combination with EGCG were assessed by using WST-8 assay; and also, Flow cytometry was utilized to identify cells undergoing apoptosis after treatments of the SW-1736 cells. We investigated the mRNA expression of genes involved in epigenetic events in SW-1736 cells by real-time qRT-PCR following the treatments. We demonstrated for the first time that the Dabrafenib–EGCG combination reduced cell viability significantly depending on concentration and induced apoptosis by 8.49-fold through investigating the additive effect together on SW-1736 cells. The IC50 doses of Dabrafenib and EGCG for 48 h were determined as 6.7 μM and 22.5 μM, respectively. The results of qRT-PCR demonstrated that the Dabrafenib–EGCG combination significantly caused the down-regulation of genes involved in epigenetic regulation. We suggest that the combination of Dabrafenib and EGCG following in vivo phase studies will contribute as an alternative treatment option for the treatment of ATC.
... Besides these examples, the range of polyphenols that have been shown to modulate major inflammatory pathways, pathway-dependent effector molecules, and gene expression is huge and include calebin A, kaempferol, anthocyanins, genistein, various phenolic acids, caffeine, allicin, cinnamon polyphenols, and many more [14,36,[119][120][121][122][123]. This demonstrates their great potential for preventing and treating the inflammation and pathogenesis of chronic diseases, and explains their great anti-inflammatory properties and anabolic effects [56,124]. ...
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Tendinitis (tendinopathy) is a pro-inflammatory and painful tendon disease commonly linked with mechanical overuse and associated injuries, drug abuse, and lifestyle factors (including poor diet and physical inactivity) that causes significant healthcare expenditures due to its high incidence. Nuclear factor kappa B (NF-kB) is one of the major pro-inflammatory transcription factors, along with other inflammation signaling pathways, triggered by a variety of stimuli, including cytokines, endotoxins, physical and chemical stressors, hypoxia, and other pro-inflammatory factors. Their activation is known to regulate the expression of a multitude of genes involved in inflammation, degradation, and cell death. The pathogenesis of tendinitis is still poorly understood, whereas efficient and sustainable treatment is missing. Targeting drug suppression of the key inflammatory regulators represents an effective strategy for tendinitis therapy, but requires a comprehensive understanding of their principles of action. Conventional monotherapies are often ineffective and associated with severe side effects in patients. Therefore, agents that modulate multiple cellular targets represent therapeutic treatment potential. Plant-derived nutraceuticals have been shown to act as multi-targeting agents against tendinitis via various anti-oxidant and anti-inflammatory mechanisms, whereat they were able to specifically modulate numerous signaling pathways, including NF-kB, p38/MAPK, JNK/STAT3, and PI3K/Akt, thus down-regulating inflammatory processes. This review discusses the utility of herbal nutraceuticals that have demonstrated safety and tolerability as anti-inflammatory agents for the prevention and treatment of tendinitis through the suppression of catabolic signaling pathways. Limitations associated with the use of nutraceuticals are also described.
... EGCG aids in declining glaucoma symptoms by inhibiting inflammatory responses by NF-kB pathway suppression in a rat glaucoma model . EGCG also directly interacts with TGFR-II, Pin1, and metalloproteinases (primarily MMP2, and MMP9, as mentioned above), which control epithelialmesenchymal transaction (EMT), EGCG-dependent inhibition of NF-kB, and cellular invasion, respectively, by its anticancerous effect (Negri et al., 2018). This illustrates that the mechanism of action of EGCG encompasses numerous signal transduction pathways. ...
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Epigallocatechin gallate (EGCG), a green tea catechin, has gained the attention of current study due to its excellent health‐promoting effects. It possesses anti‐obesity, antimicrobial, anticancer, anti‐inflammatory activities, and is under extensive investigation in functional foods for improvement. It is susceptible to lower stability, lesser bioavailability, and lower absorption rate due to various environmental, processing, formulations, and gastrointestinal conditions of the human body. Therefore, it is the foremost concern for the researchers to enhance its bioactivity and make it the most suitable therapeutic compound for its clinical applications. In the current review, factors affecting the bioavailability of EGCG and the possible strategies to overcome these issues are reviewed and discussed. This review summarizes structural modifications and delivery through nanoparticle‐based approaches including nano‐emulsions, encapsulations, and silica‐based nanoparticles for effective use of EGCG in functional foods. Moreover, recent advances to enhance EGCG therapeutic efficacy by specifically targeting its molecules to increase its bioavailability and stability are also described. The main green tea constituent EGCG possesses several health‐promoting effects making EGCG a potential therapeutic compound to cure ailments. However, its low stability and bioavailability render its uses in many disorders. Synthesizing EGCG prodrugs by structural modifications helps against its low bioavailability and stability by overcoming premature degradation and lower absorption rate. This review paper summarizes various strategies that benefit EGCG under different physiological conditions. The esterification, nanoparticle approaches, silica‐based EGCG‐NPs, and EGCG formulations serve as ideal EGCG modification strategies to deliver superior concentrations with lesser toxicity for its efficient penetration and absorption across cells both in vitro and in vivo. As a result of EGCG modifications, its bioactivities would be highly improved at lower doses. The protected or modified EGCG molecule would have enhanced potential effects and stability that would contribute to the clinical applications and expand its use in various food and cosmetic industries. EGCG is a bioactive compound present in green tea. EGCG contains pyrogallol as B‐rings while it contains an additional gallate moiety as a D‐ring, which increases the number of hydroxyl groups. The EGCG, due to galloyl moieties increasing in their hydroxyl groups, possesses more excellent antioxidant activities than EC and EGC. Considerable challenges in EGCG utilization are the low systemic bioavailability, less stability in alkaline media, temperature, high oxidative degradation, metabolic transformations, as well as toxicity at higher concentrations. Hence, some effective strategies are introduced to overcome its lower bioavailability and stability issues.
... Natural products have historically contributed to pharmacotherapy, especially for cancer and infectious diseases (5,6). (-)-Epigallocatechin-3-gallate (EGCG), is a polyphenolic component extracted from green tea, has been demonstrated to exhibit a variety of health benefits such as anti-tumor, anti-oxidant, antiinflammatory, cardiovascular protection, and neuroprotection (7,8). The recent studies revealed that EGCG inhibited proliferation and improved the sensitization of NPC cells to radiotherapy, which suggested the therapeutic potential of EGCG on NPCs (9,10). ...
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Epigallocatechin-3-gallate (EGCG), a frequently studied catechin in green tea, has been shown involved in the anti-proliferation and apoptosis of human nasopharyngeal carcinoma (NPC) cells. However, the underlying molecular mechanism of the apoptotic effects of EGCG has not been fully investigated. Recent literature emphasized the importance of Sirtuin 1 (SIRT1), an NAD+-dependent protein deacetylase, in regulating cellular stress responses, survival, and organismal lifespan. Herein, the study showed that EGCG could significantly inhibit cell proliferation and promote apoptosis of 2 NPC (CNE-2 and 5-8F) cell lines. Moreover, it was also found that SIRT1 is down-regulated by EGCG, and the SIRT1-p53 signaling pathway participates in the effects of EGCG on CNE-2 and 5-8 F cells. Taken together, the findings of this study provided evidence that EGCG could inhibit the growth of NPC cell lines and is linked with the inhibition of the SIRT1-p53 signaling pathway, suggesting the therapeutic potential of EGCG in human NPC.
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The beneficial health effects of green tea have been attributed to tea catechins. However, the molecular mechanisms of action, especially those in vivo, remain unclear. This article reviews the redox and other activities of tea catechins, using (-)-epigallocatechin-3-gallate (EGCG), as an example. EGCG is a well-known antioxidant. However, EGCG can be oxidized to generate reactive oxygen species and EGCG quinone. We propose that EGCG quinone can react with Keap-1 to activate Nrf2-regulated cytoprotective enzymes. Tissue levels of catechins are important for their biological activities; a section is devoted to reviewing the biological fates of tea catechins after ingestion. Possible EGCG oxidation in vivo and whether the oligomeric forms are biologically active in animals are discussed. We also review the effects of EGCG on the activities of enzymes, receptors, and other signaling molecules through binding and raise a question about whether the autoxidation of EGCG in vitro may lead to artifacts or misinterpretation in some studies. Finally, we discuss the challenges in the extrapolation of in vitro results to situations in vivo and the translation of laboratory studies to humans.
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Epigallocatechin gallate (EGCG) is an active catechin in green tea and has multiple biological functions, such as anti-inflammation, anti-cancer, and immune regulation. This work aimed to study the protective function of EGCG against asthma and the mechanism. Asthma in mice was induced by house dust mite (HDM) challenge. EGCG treatment alleviated tissue injury, inflammation, mucus production and collagen deposition, and it reduced M2 macrophage infiltration in mouse lung tissues induced by HDM. The bioinformatics analyses in this study suggested that target genes of EGCG were enriched in the hypoxia inducible factor-1 (HIF-1) pathway, EGCG treatment targeted HIF-1α and thereby suppressed vascular endothelial growth factor A (VEGFA) activation. Adenovirus (AAV) overexpression vectors of HIF-1α and VEGFA were administrated into mice after EGCG treatment. Either restoration of HIF-1α or VEGFA significantly blocked the protective functions of EGCG treatment against HDM-induced allergic asthma. In conclusion, this study demonstrates that EGCG treatment relieves asthmatic symptoms in mice by suppressing HIF-1α/VEGFA-mediated M2 skewing of macrophages.
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Polyphenols, as widely existing natural bioactive products, provide a vast array of advanced biomedical applications attributing to their potential health benefits that linked to antioxidant, anti-inflammatory, immunoregulatory, neuroprotective, cardioprotective function, etc. The polyphenol compounds could dynamically interact and bind with diverse species (such as polymers, metal ions, biomacromolecules, etc.) via multiple interactions, including hydrogen bond, hydrophobic, π—π, and cation—π interactions due to their unique chemical polyphenolic structures, providing far-ranging strategies for designing of polyphenol-based vehicles. Natural polyphenols emerged as multifaceted players, acting either as inherent therapeutics delivered to combat diverse diseases or as pivotal assemblies of drug delivery vehicles. In this review, we focused on the rational design and application of metal-phenolic network (MPN) based delivery systems, polyphenol-based coating films, polyphenol hollow capsules, polyphenol-incorporated hydrogels, and polymer-polyphenol-based nanoparticles (NPs) in various diseases therapeutic, including cancer, infection, cardiovascular disease, neurodegenerative disease, etc. Additionally. the versatility and mechanisms of polyphenols in the field of biomacromolecules (e.g., protein, peptide, nucleic acid, etc.) delivery and cell therapy have been comprehensively summarized. Going through the literature review, the remaining challenges of polyphenol-containing nanosystems need to be addressed are involved, including long-term stability, biosafety in vivo, feasibility of scale-up, etc., which may enlighten the further developments of this field. This review provides perspectives in utilizing natural polyphenol-based biomaterials to rationally design next generation versatile drug delivery system in the field of biomedicine, which eventually benefits public health.
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Cervical cancer is the fourth most common gynecological cancer worldwide. Although prophylactic vaccination presents the most effective method for cervical cancer prevention, chemotherapy is still the primary invasive intervention. It is urgent to exploit low-toxic natural anticancer drugs on account of high cytotoxicity and side-effects of conventional agents. As a natural product, (-)-epigallocatechingallate (EGCG) has abilities in anti-proliferation, anti-metastasis and pro-apoptosis of cervical cancer cells. Moreover, EGCG also has pharmaceutical synergistic effects with conventional agents such as cisplatin (CDDP) and bleomycin (BLM). The underlying mechanisms of EGCG suppressive effects on cervical cancer are reviewed in this article. Further research directions and ambiguous results are also discussed.
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The epimutation concept, that is, malignancy is a result of deranged patterns of gene expression due to defective epigenetic control, proposes that in the majority of adult cancers the primary (initiating) lesion adversely affects the mechanism of vertical transmission of the epigenetic pattern existing in the stem cells of differentiated tissue. Such an error-prone mechanism will result in deviant gene expression capable of accumulation at each mitosis of the affected stem cell clone. It is argued that a proportion of these proliferation products will express combinations of genes which endow them with malignant properties, such as the ability to transgress tissue boundaries and migrate to distant locations. Since the likelihood of this occurrence is dependent on the proliferation of cells manifesting the defective epigenetic transmission, the theory predicts that cancer incidence will be strongly influenced by factors regulating the turnover rate of the stem cells of the tissue in question. Evidence relating to this stipulation is examined. In addition, it would be anticipated on the basis of the selection of genes involved that the susceptibility to malignant transformation will vary according to the tissue of origin and this is also discussed.
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Notch dysregulation has been implicated in numerous tumors, including triple-negative breast cancer (TNBC), which is the breast cancer subtype with the worst clinical outcome. However, the importance of individual receptors in TNBC and their specific mechanism of action remain to be elucidated, even if recent findings suggested a specific role of activated-Notch3 in a subset of TNBCs. Epidermal growth factor receptor (EGFR) is overexpressed in TNBCs but the use of anti-EGFR agents (including tyrosine kinase inhibitors, TKIs) has not been approved for the treatment of these patients, as clinical trials have shown disappointing results. Resistance to EGFR blockers is commonly reported. Here we show that Notch3-specific inhibition increases TNBC sensitivity to the TKI-gefitinib in TNBC-resistant cells. Mechanistically, we demonstrate that Notch3 is able to regulate the activated EGFR membrane localization into lipid rafts microdomains, as Notch3 inhibition, such as rafts depletion, induces the EGFR internalization and its intracellular arrest, without involving receptor degradation. Interestingly, these events are associated with the EGFR tyrosine dephosphorylation at Y1173 residue (but not at Y1068) by the protein tyrosine phosphatase H1 (PTPH1), thus suggesting its possible involvement in the observed Notch3-dependent TNBC sensitivity response to gefitinib. Consistent with this notion, a nuclear localization defect of phospho-EGFR is observed after combined blockade of EGFR and Notch3, which results in a decreased TNBC cell survival. Notably, we observed a significant correlation between EGFR and NOTCH3 expression levels by in silico gene expression and immunohistochemical analysis of human TNBC primary samples. Our findings strongly suggest that combined therapies of TKI-gefitinib with Notch3-specific suppression may be exploited as a drug combination advantage in TNBC treatment.
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Prostate cancer is the most common cancer among men in the western world. Clinical practice is continuously challenged by the pitfalls of the available diagnostic tools. microRNAs may represent promising biomarkers in many types of human cancers, including prostate cancer. The aim of this study was to investigate microRNA expression in tumour tissue and matched plasma in a cohort of patients with primary metastatic prostate cancer. The relative expression of 12 microRNAs was assessed in diagnostic needle biopsies from the prostate and matched plasma samples in two prospective cohorts (screening cohorts) comprising 21 patients with metastatic prostate cancer and 25 control patients. An independent validation cohort of plasma samples was collected prospectively from 149 newly diagnosed patients with local/locally advanced prostate cancer. Analyses were performed using real-time polymerase chain reaction. miRNA-93 showed a significant negative correlation between expression in tumour tissue and plasma in patients with metastatic prostate cancer. Furthermore, the plasma level of miRNA-93 significantly decreased after treatment in patients with local/locally advanced prostate cancer compared to baseline plasma level. The expression of six microRNAs (let-7b, miRNA-34a, -125b, -143, -145 and -221) was downregulated, and three microRNAs (miRNA-21, -25 and miRNA-93) were upregulated in tumour tissue compared to benign prostate tissue. In plasma, six microRNAs were upregulated (miRNA-21, -125b, -126, -141, -143 and -375), while let-7b was downregulated in patients with metastatic prostate cancer compared to the control cohort. In the metastatic prostate cancer cohort, the expression of four microRNAs (miRNA-125b, -126, -143 and -221), and miRNA-141 in tissue was associated with Gleason score and prostate-specific antigen, respectively. The expression of miRNA-93 in tumour tissue was correlated with matched plasma levels and showed a significant decrease in plasma level after intervention in local prostate cancer. Differential expression between tumour and benign prostate was detected for several microRNAs in both tissue and plasma.
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Recognition and internalisation of intracellular pathogens by host cells is a multifactorial process, involving both stable and transient interactions. The plasticity of the host cell plasma membrane is fundamental in this infectious process. Here, the participation of macrophage lipid microdomains during adhesion and internalisation of the fungal pathogen Histoplasma capsulatum (Hc) was investigated. An increase in membrane lateral organisation, which is a characteristic of lipid microdomains, was observed during the first steps of Hc–macrophage interaction. Cholesterol enrichment in macrophage membranes around Hc contact regions and reduced levels of Hc–macrophage association after cholesterol removal also suggested the participation of lipid microdomains during Hc–macrophage interaction. Using optical tweezers to study cell-to-cell interactions, we showed that cholesterol depletion increased the time required for Hc adhesion. Additionally, fungal internalisation was significantly reduced under these conditions. Moreover, macrophages treated with the ceramide-glucosyltransferase inhibitor (P4r) and macrophages with altered ganglioside synthesis (from B4galnt1−/− mice) showed a deficient ability to interact with Hc. Coincubation of oligo-GM1 and treatment with Cholera toxin Subunit B, which recognises the ganglioside GM1, also reduced Hc association. Although purified GM1 did not alter Hc binding, treatment with P4 significantly increased the time required for Hc binding to macrophages. The content of CD18 was displaced from lipid microdomains in B4galnt1−/− macrophages. In addition, macrophages with reduced CD18 expression (CD18low) were associated with Hc at levels similar to wild-type cells. Finally, CD11b and CD18 colocalised with GM1 during Hc–macrophage interaction. Our results indicate that lipid rafts and particularly complex gangliosides that reside in lipid rafts stabilise Hc–macrophage adhesion and mediate efficient internalisation during histoplasmosis.
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Non‐small cell lung cancer (NSCLC) remains one of the most aggressive tumors with low life expectancy worldwide. The existence of cancer stem cells (CSCs) contributes to the failure of cancer treatment resulted from drug resistance. Altered microRNA expression has been observed in human tumors due to its role in tumor growth, progression and metastasis. Hence, the aim of our present study was to investigate the effects of miR‐485 on the CSC‐like traits in NSCLC A549‐cisplatin resistant cells and concentrate on the underlying molecular mechanism. It was found that CSC‐like phenotypes were much more enriched in A549/cisplatin (A549/CDDP) cells compared to A549‐parental cells. In addition, we observed that miR‐485 was greatly decreased in A549/CDDP cells and miR‐485 overexpression was able to decrease the stemness of A549/DDP cells. Meanwhile, epigallocatechin‐3‐gallate (EGCG), a green tea polyphenol which has been identified as an effective anticancer compound was able to increase miR‐485 expression dose‐dependently in A549/CDDP cells. Inhibitors of miR‐485 remarkably increased CSC‐like phenotypes, which could be reversed by indicated doses of EGCG. Moreover, CD44 was predicted as downstream target of miR‐485 and the correlation between them was validated by performing dual‐luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Subsequently, in vivo experiments were employed to confirm that EGCG restrained CSC‐like characteristics by increasing miR‐485 and decreasing CD44 expression. Taken together, it was implied that stemness features and CSC population were suppressed by EGCG‐modulated miR‐485/CD44 axis in A549/CDDP cells. This article is protected by copyright. All rights reserved
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In this study, antitumor activity of epigallocatechin gallate (EGCG; major component of green tea polyphenol), eugenol (active component of clove), and amarogentin (active component of chirata plant) either alone or in combination were evaluated in Hela cell line. It was evident that EGCG with eugenol-amrogentin could highly inhibit the cellular proliferation and colony formation than individual treatments. Induction of apoptosis was also higher after treatment with EGCG in combination with eugenol-amrogentin than individual compound treatments. The antiproliferative effect of these compounds was due to downregulation of cyclinD1 and upregulation of cell cycle inhibitors LIMD1, RBSP3, and p16 at G1/S phase of cell cycle. Treatment of these compounds could induce promoter hypomethylation of LimD1 and P16 genes as a result of reduced expression of DNA methyltransferase 1 (DNMT1). Thus, our study indicated the better chemotherapeutic effect of EGCG in combination with eugenol-amarogentin in Hela cell line. The chemotherapeutic effect might be due to the epigenetic modification particularly DNA hypomethylation through downregulation of DNMT1.
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Chemotherapeutic drugs exert systemic toxicity in lung cancer cells and therefore novel treatment strategies are warranted. Epigallocatechin 3-gallate (EGCG), though possessing beneficial effects in alleviating cancer, its effect has been limited due to ineffective systemic delivery, toxicity and bioavailability. To attain the maximum therapeutic response of EGCG, we have synthesized bovine serum albumin (BSA) encapsulated magnetite nanoparticle (MNPs) loaded with EGCG (nano EGCG). The synthesized nano EGCG was characterized using HR-TEM, XRD and FT-IR. Cytotoxicity analysis of BSA-MNP and nano EGCG using flow cytometry was evaluated in lung adenocarcinoma A549 cells. The effect of native and nano EGCG modulating apoptosis and Nrf2/Keap1 signaling was analysed. Nano EGCG exhibited increased ROS/RNS levels and decreased mitochondrial membrane potential, as evaluated by DCFH and JC1 staining, respectively. Expression of pro-apoptotic Bcl-2 family proteins (Bcl-2, Bax, Bak, Bim and Puma) was evaluated. This study demonstrates that native and nano EGCG induces apoptosis through the involvement of ROS leading to loss in mitochondrial membrane potential. EGCG exhibited an increased expression of Nrf2 and Keap1 that could regulate apoptosis in A549 cells. This study, for the first time reveals the potential of BSA-MNPs loaded EGCG as drug target and renders better efficacy against lung cancer cells.
Article
Objective: To investigate the role of phosphatidylinositol-3-kinase protein kinase B (PI3K/Akt) signaling pathway in the apoptosis of H1299 lung cancer cells induced by epigallocatechin gallate (EGCG). Materials and methods: H1299 lung cancer cells were treated with EGCG at a dose of 10 µM, 20 µM, and 40 µM, respectively. Cell culture was performed for 72 h and then: 1, cell proliferation was detected by MTT assay; 2, cell apoptosis rate was detected by flow cytometry; 3, expression of Caspase-3, Bax, and Bcl-2 was detected by Western blot; 4, expression of PI3K, p-PI3K, Akt, and p-Akt was detected by Western blot. Results: The proliferation of H1299 cells was significantly inhibited 72 h after treatment with different doses of EGCG, and cell apoptosis rate was significantly increased (p<0.05). Compared with those in the control group, expression of PI3K and Akt in the lung cancer cells H1299 after EGCG treatment showed no significant differences (p>0.05), while expression levels of p-PI3K and p-Akt were significantly reduced (p<0.05). Conclusions: EGCG can inhibit the proliferation and induce apoptosis of H1299 lung cancer cells, and the effect is related to the inhibition of the activation of PI3K/Akt signaling pathway.
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The tumor microenvironment is complex with the cancer stem cell (CSC) as a member within its community. This population possesses the capacity to self-renew and to cause cellular heterogeneity of the tumor. CSCs are resistant to conventional anti-proliferative drugs. In order to be curative, it is imperative that CSCs must be eliminated by cancer therapy. A variety of dietary phytochemicals and repositioned drugs can act synergistically with conventional anti-cancer agents. In this review, we advocate the development of a novel approach, namely combination therapy by incorporating both phytochemicals and repositioned drugs to target CSCs. We cover select dietary phytochemicals (curcumin, resveratrol, EGCG, genistein) and repurposed drugs (metformin, niclosamide, thioridazine, chloroquine). Five of the eight (curcumin, resveratrol, EGCG, genistein, metformin) are listed in "The Halifax Project", that explores "the concept of a low-toxicity 'broad-spectrum' therapeutic approach that could simultaneously target many key pathways and mechanisms" [1]. For these compounds, we discuss their mechanisms of action, in which models their anti-CSC activities were identified, as well as advantages, challenges and potentials of combination therapy.