ArticlePDF Available

Artemisia absinthium (AA): A novel potential complementary and alternative medicine for breast cancer

  • Manipal Academy of Higher Education - Dubai Campus

Abstract and Figures

Natural products have become increasingly important in pharmaceutical discoveries, and traditional herbalism has been a pioneering specialty in biomedical science. The search for effective plant-derived anticancer agents has continued to gain momentum in recent years. The present study aimed to investigate the role of crude extracts of the aerial parts of Artemisia absinthium (AA) extract in modulating intracellular signaling mechanisms, in particular its ability to inhibit cell proliferation and promote apoptosis in a human breast carcinoma estrogenic-unresponsive cell line, MDA-MB-231, and an estrogenic-responsive cell line, MCF-7. Cells were incubated with various concentrations of AA, and anti-proliferative activity was assessed by MTT assays, fluorescence microscopy after propidium iodide staining, western blotting and cell cycle analysis. Cell survival assays indicated that AA was cytotoxic to both MDA-MB-231 and MCF-7 cells. The morphological features typical of nucleic staining and the accumulation of sub-G1 peak revealed that the extract triggered apoptosis. Treatment with 25 μg/mL AA resulted in activation of caspase-7 and upregulation of Bad in MCF-7 cells, while exposure to 20 μg/mL AA induced upregulation of Bcl-2 protein in a time-dependent response in MDA-MB-231 cells. Both MEK1/2 and ERK1/2 was inactivated in both cell lines after AA treatment in a time-dependent manner. These results suggest that AA-induced anti-proliferative effects on human breast cancer cells could possibly trigger apoptosis in both cell lines through the modulation of Bcl-2 family proteins and the MEK/ERK pathway. This might lead to its possible development as a therapeutic agent for breast cancer following further investigations.
Content may be subject to copyright.
Artemisia absinthium (AA): a novel potential complementary
and alternative medicine for breast cancer
Gowhar Shafi Tarique N. Hasan Naveed Ahmed Syed
Amal A. Al-Hazzani Ali A. Alshatwi
A. Jyothi Anjana Munshi
Received: 16 May 2011 / Accepted: 25 January 2012
ÓSpringer Science+Business Media B.V. 2012
Abstract Natural products have become increasingly
important in pharmaceutical discoveries, and traditional her-
balism has been a pioneering specialty in biomedical science.
The search for effective plant-derived anticancer agents has
continued to gain momentum in recent years. The present
study aimed to investigate the role of crude extracts of the
aerial parts of Artemisia absinthium (AA) extract in modu-
lating intracellular signaling mechanisms, in particular its
ability to inhibit cell proliferation and promote apoptosis in a
human breast carcinoma estrogenic-unresponsive cell line,
MDA-MB-231, and an estrogenic-responsive cell line, MCF-
7. Cells were incubated with various concentrations of AA,
and anti-proliferative activity was assessed by MTT assays,
fluorescence microscopy after propidium iodide staining,
western blotting and cell cycle analysis. Cell survival assays
indicated that AA was cytotoxic to both MDA-MB-231 and
MCF-7 cells. The morphological features typical of nucleic
staining and the accumulation of sub-G1 peak revealed that
the extract triggered apoptosis. Treatment with 25 lg/mL AA
resulted in activation of caspase-7 and upregulation of Bad in
MCF-7 cells, while exposure to 20 lg/mL AA induced
upregulation of Bcl-2 protein in a time-dependent response in
MDA-MB-231 cells. Both MEK1/2 and ERK1/2 was inacti-
vated in both cell lines after AA treatment in a time-dependent
manner. These results suggest that AA-induced anti-prolif-
erative effects on human breast cancer cells could possibly
trigger apoptosis in both cell lines through the modulation of
Bcl-2 familyproteinsand the MEK/ERK pathway.This might
lead to its possible development as a therapeutic agent for
breast cancer following further investigations.
Keywords Artemisia absinthium Caspase-7 Bad
Bcl-2 MEK/ERK MDA-MB-231 MCF-7
Apoptosis Cancer therapy
Artemisia absinthium (AA) is commonly called worm-
wood, and is locally known as ‘Tethwen’ in the Kashmir
Valley, India. It is used in indigenous medicine as a ver-
mifuge, an insecticide, an antispasmodic, an antiseptic, and
in the treatment of chronic fevers and inflammation of the
liver [1]. Its essential oil has antimicrobial [2] and anti-
fungal activity [3]. Chemical analysis of AA extracts has
shown that its volatile oil is rich in thujone, which has been
reported as an anthelmintic [4]. In Turkish folk medicine,
AA has been used as an antipyretic, antiseptic, anthel-
mintic, tonic, diuretic, and for the treatment of stomach-
aches [5].
Breast cancer is the most common oncological disease in
women worldwide, and its incidence and mortality rates
G. Shafi A. Jyothi A. Munshi (&)
Department of Molecular Biology, Institute of Genetics and
Hospital for Genetic Diseases, Osmania University, Begumpet,
Hyderabad 500016, Andhra Pradesh, India
G. Shafi A. Munshi
Dr. NTR University of Health Sciences, Vijayawada,
Andhra Pradesh, India
G. Shafi T. N. Hasan N. A. Syed A. A. Alshatwi
A. Munshi
Molecular Cancer Biology Research Lab (MCBRL),
Department of Food Sciences and Nutrition, College of Food
and Agricultural Sciences, King Saud University, Riyadh,
Saudi Arabia
A. A. Al-Hazzani
Department of Botany and Microbiology,
King Saud University, Riyadh, Saudi Arabia
Mol Biol Rep
DOI 10.1007/s11033-012-1569-0
may be explained by differences in the relative risk or
prevalence of risk factors, including dietary factors [6].
However, the efficacy of currently available drugs is very
limited, and anticancer agents that can target multiple points
in the apoptotic cascade to achieve synergistic actions are
urgently required. Chinese herbs have obtained considerable
attention for the prevention and treatment of certain cancer
types in clinical studies [710]. In many cases however,
extracts obtained from plants are not highly effective and
require chemical modification for improved potency and
toxicity profile [1113]. Several phytochemicals that have
been used in clinical cancer chemotherapy were originally
derived from herbs and plants, such as paclitaxel [9,14],
etoposide [15], camptothecin [8] and vinca alkaloids [16].
Thus, studies of naturally-occurring plant-based agents
could lead to the development of new strategies for the
management of cancer and related diseases. Specific com-
pounds in certain foods have been shown to reduce human
breast cancer cell proliferation through apoptosis and cell
cycle arrest [17]. Interestingly, the apoptosis pathway was a
novel target for cancer chemoprevention in recent studies of
several chemopreventive agents [18]. The current study was
designed to investigate the in vitro anticancer activity of
crude extracts of the aerial parts of AA on estrogen-
responsive MCF-7 and estrogen-unresponsive MDA-MB-
231 human breast cancer cell lines.
Materials and methods
Preparation of plant material
Plants were selected on the basis of ethnopharmacology,
and 3 kg of plant material was collected locally. It was
identified by a botanist from the Department of Botany,
Osmania University, and a specimen was deposited at the
herbarium. The aerial parts of AA were air-dried in shade,
then powdered using a milling machine. The powdered
plant material was extracted with methanol as described
previously [19]. Briefly, powdered plant material was
soaked in ten times the volume of methanol for extraction.
Extraction was performed three times, and each extraction
was performed for 24 h. Methanolic filtrate was then
evaporated under reduced pressure to obtain a residue
(500 g of AA yielded 29.15 g of residue). The residue was
dried using a rotary evaporator to obtain the powder/paste,
and the required quantity was dissolved in dimethyl sulf-
oxide (DMSO).
Preparation of drug
A stock of plant extract was prepared to a concentration of
1 mg/mL in DMSO and sterilized by autoclaving at 121°C
and 15 lb for 15 min. Then, five concentrations of test
drug (5, 20, 50, and 100 lg/mL) were prepared by dilut-
ing stock with DMEM. DMEM alone was used as a
vehicle control.
Maintenance of cell lines
MDA-MB-231 and MCF-7 cells were procured from the
National Centre for Cell Science (Pune, India). The cell
lines were maintained and propagated in 90% Dulbecco’s
Modified Eagle’s Medium (DMEM) supplemented with
10% fetal bovine serum (FBS) and 1% penicillin/strepto-
mycin. Cells were cultured as adherent monolayers to
approximately 70–80% confluence, and maintained at 37°C
in a humidified atmosphere of 5% CO
. Cells were har-
vested after brief trypsinization. All chemicals used were of
research grade.
Cell toxicity and viability assays
MDA-MB-231 and MCF-7 cells were grown in DMEM at
37°C under 5% CO2 in a humidified incubator. Cells were
harvested, counted and transferred to 96-well plates, and
incubated for 24 h prior to addition of the test compounds.
The extracted compounds were processed and applied in
various concentrations, and the treated cells were incubated
for 72 h. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-
tetrazolium bromide) (5 mg) was dissolved in 1 mL
phosphate-buffered saline (PBS), and 25 lL MTT solution
was added to each of the wells. The plates were wrapped in
aluminum foil and incubated at 37°C for 4 h. The solution
in each well, containing media, unbound MTT and dead
cells, was removed by suction, and replaced with 200 lL
DMSO. The plates were then shaken, and the optical
density was measured using a microplate reader at 575 nm.
Three independent experiments were performed for each
study and all measurements were performed in triplicate.
Results were expressed as the percentage proliferation with
respect to vehicle-treated cells. Cells were further treated
with IC
concentrations of AA (25 lg/mL for MCF-7
cells and 20 lg/mL for MDA-MB-231 cells) or vehicle
(0.1%) for 24, 48 or 72 h.
Apoptotic assays
The apoptotic effects of AA on MDA-MB-231 and MCF-7
cells were analyzed by nuclear DNA staining assays. AA-
treated and untreated cells were fixed in 4% paraformal-
dehyde in PBS for 20 min, washed twice with PBS, and
stained with 1 lg/mL propidium iodide (PI) (Sigma) for
15 min. Stained cells were then washed twice with PBS.
The changes in nuclei were observed under an ultraviolet
fluorescent microscope (Carl Zeiss).
Mol Biol Rep
Flow cytometry analysis
MDA-MB-231 and MCF-7 cells in the exponential phase
of growth were treated with AA extract (20 lg/mL for
MDA-MB-231 cells and 25 lg/mL for MCF-7 cells) for 24
and 48 h, then harvested by trypsinization, washed twice
with ice-cold PBS and fixed by 70% ethanol at -20°C for
at least 30 min. The fixed cells were then washed twice
with ice-cold PBS and stained with 50 lg/mL PI for
30 min. Cell cycle distribution was analyzed by using a
FACSCalibur (Becton–Dickinson, USA). Data from
10,000 cells per sample were collected and analyzed using
the Cell Fit Cell analysis program.
Western blot analysis
MDA-MB-231 and MCF-7 cells were grown in 6-well
plates, and when cell density reached 80–90% confluence,
cells were treated with AA (20 lg/mL for MDA-MB-231
cells and 25 lg/mL for MCF-7 cells) for 24, 48 or 72 h.
After treatment, cells were collected and washed twice
with cold PBS. The cells were then lysed in lysis buffer
(50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1% Nonidet
P-40, 2 mM EDTA, 1 mM EGTA, 1 mM NaVO3, 10 mM
NaF, 1 mM DTT, 1 mM PMSF, 25 lg/mL aprotinin, and
25 lg/mL leupeptin) and kept on ice for 30 min. The lysates
were then centrifuged at 12,0009gat 4°Cfor20minand
the supernatants were stored at -70°C until use. Protein
concentration was determined by the Bradford method.
Aliquots of the lysates (30 lgofprotein)wereseparatedin
12% SDS-PAGE and transferred onto a nitrocellulose
membrane using transfer buffer (192 mM glycine, 25 mM
Tris–HCl, pH 8.8, and 20% methanol [v/v]). After blocking
with 5% non-fat dried milk, the membranes were subse-
quently incubated with the corresponding primary antibod-
ies, as indicated: a rabbit anti-PARP p85 fragment, anti-
active ERK1/2, anti-active MEK1/2 and anti-MEK1/2
polyclonal antibody, a mouse anti-caspase-7, anti-Bcl-2 and
anti-Bad monoclonal antibody, rabbit anti-ERK 1/2 poly-
clonal antibody (Abcam Inc, USA); Antibody recognition
was detected with the respective secondary antibody, anti-
mouse IgG antibodies linked to horseradish peroxidase
(Abcam Inc, USA). Primary antibodies were used at a
1:1,000 dilution, while horseradish peroxidase-conjugated
horse anti-rabbit IgG (Sigma Chemicals, USA) was used at a
1:5,000 dilution as the secondary antibody. The membrane
was then exposed and protein bands were detected using
Enhanced Chemiluminescence (Abcam Inc, USA).
Statistical analysis
The data were expressed as the mean (±SD). Comparisons
between groups were performed by Student’s ttest and
one-way analysis of variance (ANOVA). All statistical
analyses were performed using the statistical software
PASW 18.0. Pvalues of \0.05 were regarded as statisti-
cally significant.
Inhibition of cell proliferation in AA-treated MCF-7
and MDA-MB-231 cells
To investigate the effects of AA methanolic extract on
MDA-MB-231 and MCF-7 cell proliferation, the cells were
treated with various concentrations of AA for 72 h. AA at
25 lg/mL caused almost 50% inhibition of cell prolifera-
tion in MCF-7 cells compared with controls, while 20 lg/
mL caused 50% inhibition in MDA-MB-231 cells with
(Figs. 1,2a, b). The anti-proliferative activity was time-
dependent in both cell types. Therefore, we used respective
concentrations to perform the downstream experiments.
Microscopic signs of apoptosis in AA-treated cells
The ability of AA to alter cell morphology indicative of
apoptosis in the cancer cell lines was assessed using PI
staining (Fig. 3). After incubation for 48 h with AA, the
cells became rounded in appearance, exhibited nuclear
condensation and significant nuclei fragmentation indicat-
ing apoptosis.
Flow cytometry analysis
Cell cycle analysis by flow cytometry was used to confirm
the AA-induced cell death in MDA-MD-231 and MCF-7
Fig. 1 Cell viability was determined by MTT assays. Cells were
treated with various concentrations of AA, and results are expressed
as percentages of proliferation compared with untreated control
(mean ±SD, n=3)
Mol Biol Rep
cells. Indeed, both cell lines accumulated in the sub-G1
phase gradually from 24 to 72 h after treatment with AA,
whereas the number of cells in the G2/M phase decreased
in a similar fashion. These results indicate that apoptosis
was induced in MDA-MD-231 (Fig. 4a) and MCF-7 cells
(Fig. 4b).
Effects of AA on PARP cleavage and caspase-7
MDA-MB-231 and MCF-7 cells were treated with 25 and
20 lg/ml AA, respectively, and were harvested after 24, 48
Fig. 2 Typical effects of AA in reducing the viability of MCF-7
(a) and MDA-MB-231 cells (b). Cells were treated with 25 lg/mL of
AA for 24, 48 and 72 h (mean ±SD, n=3)
Fig. 3 Detection of apoptotic morphological changes in MDA-MB-
231 and MCF-7 cells treated with AA for 72 h. Nuclei were stained
with PI and examined by fluorescence microscopy. Untreated MCF-7
cells (left) and MCF-7 cells treated with 25 lg/mL AA (right);
untreated MDA-MB-231 cells (left) and MDA-MB-231 cells treated
with 20 lg/mL AA (right)
Fig. 4 MDA-MB-231 (a) and MCF-7 (b) cells were treated with 20
and 25 lg/mL AA, respectively for 24, 48 and 72 h. Cell cycle
distribution was determined in samples stained with propidium iodide
and measured by flow cytometry; percentage of cells in sub-G1, G1/S,
and G2/M phase was then calculated. Data shown are from a
representative experiment repeated three times
Mol Biol Rep
and 72 h of exposure. Caspase-7 cleavage was detected
after 24 h of exposure to AA, and PARP was cleaved to its
active 85 kDa form in both cell lines (Fig. 5a, b). These
results showed that AA induced apoptosis through the
cleavage of PARP, a substrate of several interleukin-1b-
converting enzyme-like proteases.
Modulation of regulatory apoptotic proteins Bad
and Bcl-2
Cell lines were treated with their respective IC
tration of AA, harvested after 24, 48 and 72 h of exposure,
and analyzed as described above. Pro-apoptotic Bad was
upregulated in MCF-7 cells treated for 24 h (Fig. 5c) but
anti-apoptotic Bcl-2 protein was decreased in MDA-MB-
231 cells after treatment (Fig. 5d). No changes were
observed Bcl-2 protein levels in MCF-7 cells or Bad pro-
tein levels in MBA-MB-231 cells after treatment (data not
included). The results suggest that AA-induced apoptosis
in human breast cancer cells might be mediated through the
modulation of the Bcl-2/Bad pathway.
Effects of AA on the MEK/ERK pathway
The involvement of mitogen-activated protein kinases
(MAPKs) in AA-induced apoptosis in MCF-7 and MDA-
MB-231 cells was analyzed by assessing the time-depen-
dent effect on the ERK1/2 MAPK pathway. As shown in
Fig. 6a, AA-induced inactivation of ERK1/2 began at 48 h
and lasted up to 72 h in MCF-7 cells. In contrast, phospho-
ERK1/2 was slowly downregulated by AA over 72 h in
MDA-MB-231 cells (Fig. 6b). The same blots were sub-
sequently stripped and re-blotted with an antibody that
recognized total ERK to verify equal amounts of the pro-
tein in various samples. MEK1/2 phosphorylation was
decreased in a time-dependent fashion following AA
treatment over the same timeframe, as seen for phosphor-
ylated ERK1/2 (Fig. 6c, d). Given that ERK1/2 activation
is considered to be a determining factor in cell survival and
apoptosis, these results suggest that AA-induced apoptosis
in both cell lines may be associated with activation of the
MEK/ERK pathway.
The drug discovery process is becoming more complex and
capital-intensive, and systematic and critical review of the
methods and approach towards the entire process is
required to rediscover the discovery process afresh.
Screening of medicinal plants for potential anticancer
properties has increased greatly over the past few decades.
For instance, the US National Cancer Institute has imple-
mented a large-scale project of acquisition and screening of
compounds isolated from medicinal plants originating from
various regions of the world. These medicinal plants are
identified based on ethnopharmacological, chemosystemic
and ecological information. In the present study, irrevers-
ible inhibition of proliferation was observed in AA-treated
Fig. 5 Effects of AA on cleaved PARP and caspase-7 proteins.
MCF-7 cells were treated with 25 lg/mL AA (a) and MDA-MB-231
cells were treated with 20 lg/mL AA (b) for indicated times. Total
cell lysates were prepared and western blotting was performed with
antibodies specific for corresponding proteins. Effects of AA on Bad
protein; MCF-7 cells were treated with 25 lg/mL AA for indicated
times (c). Effects of AA on Bcl-2 protein; MDA-MB-231 cells were
treated with 20 lg/mL AA for indicated times (d)
Fig. 6 Effects of AA on the levels of ERK1/2, pERK1/2, MEK1/2
and pMEK1/2 in MCF-7 (a,c) and MDA-MD-231 (b,d) cells. Cells
were cultured as described in the ‘Materials and methods’ section.
MCF-7 and MDA-MB-231 cells were treated with 25 and 20 lg/mL
AA, respectively, for 24, 48 and 72 h. Cells were harvested and total
cell lysates were prepared. Western blotting was performed with
antibodies specific for corresponding proteins. Experiments were
repeated three times with similar results
Mol Biol Rep
breast cancer cells in a concentration and time-dependent
fashion. Thus, it is clear that AA inhibits cell proliferation
via apoptotic death, as visualized by nuclear fragmentation
and condensation, as well as increased sub-G1 phase.
Apoptosis is an active physiological process resulting in
cellular self-destruction that involves specific morphological
and biochemical changes in the nucleus and cytoplasm [20].
Agents that suppress the proliferation of malignant cells by
inducing apoptosis may represent a useful mechanistic
approach to both cancer chemoprevention and chemotherapy.
While many anticancer agents have been developed, unfa-
vorable side-effects and resistance are serious problems [21].
Thus, there is growing interest in the use of plant materials for
the treatment of various cancers, and for the development of
safer and more effective therapeutic agents [22]. AA has been
used as a folk remedy for several diseases without any
understanding of the underlying mechanisms. Therefore, we
chose to investigate the role of AA in the inhibition of pro-
liferation and promotion of apoptosis in human breast cancer
MDA-MB-231 and MCF-7 cells. AA treatment induced the
activation of caspase-7 in these cell lines. An inactive caspase-
7 precursor was cleaved to form the active protease during
apoptosis, resulting in PARP degradation.
In general, apoptosis is controlled by the complex inter-
play between regulatory proteins of the Bcl-2 family [23].
These pro- and anti-apoptotic proteins are key regulators of
the intrinsic apoptotic pathway, controlling the point of no
return and setting the threshold for engagement of the death
machinery [24]. Previous reports have shown that the ratio
of Bax to Bcl-2 determines, in part, the susceptibility of cells
to death signals [25]. Therefore, Bcl-2 proteins have
emerged as attractive targets for the development of novel
anticancer drugs [26]. Changes in the Bcl-2/Bax ratio have
been reported to be caused by downregulation of Bcl-2 and
slight downregulation of Bax [27], downregulation of Bcl-2
and upregulation of Bax [28,29], and downregulation of
Bcl-2 with no change in the level of Bax [30]. However,
other members of the Bcl-2 family, Bax, Bad, Bak, Bik and
Bid, promote cell death [31]. Anti-apoptotic Bcl-2 expres-
sion was significantly downregulated after AA exposure for
48 h in MDA-MB-231 cells. However, our results clearly
showed upregulation of Bad after treatment of MCF-7 for
24 h. Thus it is reasonable to conclude that AA treatment
may induce apoptosis by regulating the Bcl-2 family.
In addition, MAPK cascades transmit and amplify sig-
nals involved in cell proliferation and apoptosis. Three
major MAPK pathways exist in human tissues, but the
ERK1/2 cascade is most relevant to breast cancer because
if the appropriate receptors are present, upregulation of
ligands may be responsible for increased MAPKs in these
cancers [32]. Exposure of MDA-MB-231 and MCF-7 cells
to AA resulted in highly significant inhibition of MEK and
ERK1/2. The mechanism of action of many anticancer
drugs is based on their ability to induce apoptosis [33,34].
Hence, we were interested in identifying if cancer cells
treated with methanolic extracts of AA utilized apoptosis
as their mode of cell death. This was approached by
studying distinct morphological features (nuclear chroma-
tin condensation, fragmentation of nuclear material) and
molecular features (expression of certain crucial genes).
The MAPK pathway represents a cascade of phosphor-
ylation events including three pivotal kinases, namely Raf,
MEK and ERK; activated MEK1/2 can activate ERK1/2.
This concurs with the observation of Agarwal et al. [35]
who showed that a polyphenolic fraction isolated from
grape seeds induced growth inhibition of breast MDA-MB-
468 carcinoma cells by inhibiting MAPK activation. Taken
together, we therefore suggest that AA plays critical roles
in apoptotic and MAPK pathways. The benefit of herbal
decoctions is that they can nourish the body as a whole by
supporting various organ systems. More importantly as
they have no synthetic elements, there is very less likeli-
hood of an unexpected adverse effect.
In conclusion, AA inhibits proliferation of human breast
cancer MDA-MB-231 and MCF-7 cells through the
induction of apoptosis by regulating Bcl-2 family proteins
and MEK/MAPK signaling. We also found in this study
that p53-independent cell death induced by AA is regulated
Fig. 7 Schematic summary of our findings. On the basis of the results
of the current study, we propose that AA treatment triggers activation
of the MEK/ERK pathway, which then initiates the mitochondrial
pathway of caspase activation, and regulates Bad and Bcl-2 family
proteins, culminating in the apoptotic death of MCF-7 and MDA-MB-
231 cells
Mol Biol Rep
through an MEK–ERK–mitochondria–caspase cascade that
may be critical to improvement of the clinical outcome
(Fig. 7). This suggests AA extract has an anticancer effect
that is mediated via the apoptotic pathway in human breast
cancer cell lines. However, the potent extracts need to be
tested on several other cancer cell lines, and animal models
should be used to establish the therapeutic efficacy and
toxicity of this substance. In addition, extracts must be
subjected to HPLC and GC–MS analyses to identify and
characterize the efficacious phytotherapeutic and/or bio-
active compound(s) in AA.
1. Koul MK (1997) Medicinal Plants of Kashmir and Ladakh,
Temperate and Cold Arid Himalaya, Indus Publishing Company,
FS-5, Tagore Garden, New Delhi p 102, ISBN: 81-7837-061-6
2. Juteau F, Jerkovic I, Masotti V, Milos M, Mastelic J, Bessiere
JM, Viano J (2003) Composition and anti-microbial activity of
essential oil of Artemisia absinthium from Croatia and France.
Planta Med 69:158–161
3. Saban K, Recep M, Ahmet CAA, Ali Y (2005) Determination of
the chemical composition and antioxidant activity of the essential
oil of Artemisia dracunculus and of the antifungal and antibac-
terial activities of Turkish Artemisia absinthium,Artemisia dra-
cunculus,Artemisia santonicum, and Artemisia spicigera
essential oils. J Agric Food Chem 53:9452–9458
4. Meschler JP, Howlett AC (1999) Thujone exhibits low affinity for
cannabinoid receptors but fails to evoke cannabimimetic
responses. Pharm Biochem Behav 62:413–480
5. Baytop (1984) Therapy with Medicinal Plants in Turkey. Istanbul
University Press, Istanbul, pp 166–167
6. Jo EH, Kim SH, Ra JC, Kim SR, Cho SD, Jung JW et al (2005)
Chemopreventive properties of the ethanol extract of Chinese
licorice (Glycyrrhiza uralensis) root: induction of apoptosis and
G1 cell cycle arrest in MCF-7 human breast cancer cells. Cancer
Lett 230:239–247
7. Wu CC, Chan ML, Chen WY, Tsai CY, Chang FR, Wu YC
(2005) Pristimerin induces caspase-dependent apoptosis in MDA-
MB-231 cells via direct effects on mitochondria. Mol Cancer
Ther 4:1277–1285
8. Garcia-Carbonero R, Supko JG (2002) Current perspectives on
the clinical experience, pharmacology, and continued develop-
ment of the camptothecins. Clin Cancer Res 8:641–661
9. Rowinsky EK, Donehower RC (1995) Paclitaxel (taxol). N Engl J
Med 332:1004–1014
10. Zhu JY, Lavrik IN, Mahlknecht U, Giaisi M, Proksch P, Kram-
mer PH, Li- Weber M (2007) The traditional Chinese herbal
compound rocaglamide preferentially induces apoptosis in leu-
kemia cells by modulation of mitogen-activated protein kinase
activities. Int J Cancer 121:1839–1846
11. Huang M, Gao H, Chen Y, Zhu H, Cai Y, Zhang X, Miao Z, Jiang
H, Zhang J, Shen H, Lin L, Lu W, Ding J (2007) Chimmitecan, a
novel 9-substituted camptothecin, with improved anticancer
pharmacologic profiles in vitro and in vivo. Clin Cancer Res
12. Tamvakopoulos C, Dimas K, Sofianos ZD, Hatziantoniou S, Han
Z, Liu ZL, Wyche JH, Pantazis P (2007) Metabolism and anti-
cancer activity of the curcumin analogue, dimethoxycurcumin.
Clin Cancer Res 13:1269–1277
13. Li L, Gao Y, Zhang L, Zeng J, He D, Sun Y (2008) Silibinin
inhibits cell growth and induces apoptosis by caspase activation,
downregulating surviving and blocking EGFR-ERK activation in
renal cell carcinoma. Cancer Lett 272:61–69
14. Razis ED, Fountzilas G (2001) Paclitaxel: epirubicin in meta-
static breast cancer–a review. Ann Oncol 12:593–598
15. Hainsworth JD, Greco FA (1995) Etoposide: twenty years later.
Ann Oncol 6:325–341
16. Jordan MA, Thrower D, Wilson L (1991) Mechanism of inhibi-
tion of cell proliferation by Vinca alkaloids. Cancer Res 51:
17. Ganry O (2002) Phytoestrogen and breast cancer prevention. Eur
J Cancer Prev 11:519–522
18. Sun SY, Hail N, Lotan R (2004) Apoptosis as a novel target for
cancer chemoprevention. J Natl Cancer Inst 96:662–672
19. Shafi G, Munshi A, Hasan TN, Alshatwi AA, Jyothy A, Lei DKY
(2009) Induction of apoptosis in HeLa cells by chloroform frac-
tion of seed extracts of Nigella sativa. Cancer Cell Int 9:29
20. Mans DRA, Da Rocha BA (2000) Schwartsmann G. Anti-cancer
drug discovery and development in Brazil: target plant as a
rational strategy to acquire candidate anti-cancer compounds. The
Oncologist 5:185–198
21. Khan MR, Mlungwana SM (1999) c-sitosterol, a cytotoxic sterol
from Markhamia zanzibarica and Kigelia africana. Fitoterapia
22. Panchal RG (1998) Novel therapeutic strategies to selectively kill
cancer cells. Biochem Pharmacol 55:247–252
23. Ramos S (2007) Effects of dietary flavonoids on apoptotic
pathways related to cancer chemoprevention. J Nutr Biochem
24. Williams GT, Smith CA (1993) Molecular regulation of apop-
tosis: Genetic controls on cell death. Cell 74:777–779
25. Marzo I, Naval J (2008) Bcl-2 family members as molecular
targets in cancer therapy. Biochem Pharmacol 76:939–946
26. Chang J, Hsu Y, Kuo P, Kuo Y, Chiang L, Lin C (2005) Increase
of Bax/Bcl-XL ratio and arrest of cell cycle by luteolin in
immortalized human hepatoma cell line. Life Sci 76:1883–1893
27. Mohammad R, Giri A, Goustin AS (2008) Small-molecule
inhibitors of Bcl-2 family proteins as therapeutic agents in cancer.
Recent Pat Anti-cancer Drug Discov 3:20–30
28. Cha YY, Lee EO, Lee HJ, Park YD, Ko SG, Kim DH et al (2004)
Methylene chloride fraction of Scutellaria barbata induces
apoptosis in human U937 leukemia cells via the mitochondrial
signaling pathway. Clin Chim Acta 348:41–48
29. Paris C, Bertoglio J, Breard J (2007) Lysosomal and mitochon-
drial pathways in milterfosine-induced apoptosis in U937 cells.
Apoptosis 12:1257–1267
30. Han MH, Yoo YH, Choi YH (2008) Sanguinarine-induced
apoptosis in human leukemia U937 cells via Bcl-2 downregula-
tion and caspase-3 activation. Chemotherapy 54:157–165
31. Reed JC (1997) Double identity for proteins of the Bcl-2 family.
Nature 387:773–776
32. Santen RJ, Song RX, McPherson R, Kumar R, Adam L, Jeng
MH, Yue W (2002) The role of mitogen-activated protein (MAP)
kinase in breast cancer. J Steroid Biochem Mol Biol 80:239–256
33. Motomura M, Kwon KM, Suh SJ, Lee YC, Kim YK, Lee IS
(2008) Propolis induces cell cycle arrest and apoptosis in human
leukemic U937 cells through Bcl-2/Bax regulation. Environ
Toxicol Pharmacol 26:61–67
34. Sen S, D’Incalci M (1992) Biochemical events and relevance to
cancer chemotherapy. FEBS Lett 307:122–127
35. Agarwal C, Sharma Y, Zhao J, Agarwal R (2000) A polyphenolic
fraction from grape seeds causes irreversible growth inhibition of
breast carcinoma MDA-MB468 cells by inhibiting mitogen-
activated protein kinases activation and inducing G1 arrest and
differentiation. Clin Cancer Res 6:2921–2930
Mol Biol Rep
... It is also cultivated in Mongolia, Korea, Russia's Far East and Japan. As a traditional Chinese herbal medicine, FAA has antipyretic, analgesic, and hemostatic effects (10). For thousands of years, it has been used internally to warm channels, arrest bleeding, dispel cold, and relieve pain, and is applied externally to eliminate dampness and relieve itching (11). ...
... According to pharmacology research, FAA contains multiple active chemical constituents, such as flavonoids, terpenoids, phenolic acids, and volatile oils (15,16), and exhibits a variety of effects, including anticancer, antiinflammation, and anti-oxidation (17,18). For example, Shafi et al. suggested that FAA inhibited the proliferation and promoted apoptosis in breast cancer cells through Bcl-2 family proteins and the MEK/ERK pathway (10). It was also reported that FAA exhibited a dose-dependent inhibitory effect on hepatoma cells (11). ...
... It was also reported that FAA exhibited a dose-dependent inhibitory effect on hepatoma cells (11). However, although many studies have verified that FAA exerts remarkable antitumor functions, the underlying mechanisms have not yet been comprehensively understood (10,11,17,18). ...
Full-text available
Background: Folium Artemisia argyi (FAA) is a traditional Chinese herbal medicine that is widely used in the clinic. However, the underlying mechanisms of its anticancer effects have not been fully elucidated. Methods: In this study, we applied a network pharmacology approach to identify the potential mechanisms of FAA against breast cancer. To be specific, we screened the active ingredients and potential targets of the FAA through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Meanwhile, we employed the oral bioavailability (OB) and drug-likeness (DL) to search for potential bioactive compounds of FAA. Breast cancer-related target genes data were gathered from the GeneCards and Online Mendelian Inheritance in Man (OMIM) databases, and the protein-protein interaction (PPI) data were acquired from the Search Tool for the Retrieval of Interacting Genes (STRING) database. In addition, we constructed the network and performed Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Enrichment Analysis. Results: We obtained a total of nine active ingredients and 236 potential targets from FAA to construct a network, which showed that quercetin served as the major ingredient in FAA. AKT1 (RAC-alpha serine/threonine-protein kinase), MYC (Myc proto-oncogene protein), CASP3 (Caspase-3), EGFR (Epidermal growth factor receptor), JUN (Transcription factor AP-1), CCND1 (G1/S-specific cyclin-D1), VEGFA (Vascular endothelial growth factor A), ESR1 (Estrogen receptor), MAPK1 (Mitogen-activated protein kinase 1), and EGF (pro-epidermal growth factor) were identified as key targets of FAA in the treatment of breast cancer. The PPI cluster demonstrated that AKT1 was the seed in this cluster, indicating that AKT1 played a crucial role in connecting other nodes in the PPI network. This enrichment demonstrated that FAA was highly related to signal transduction, endocrine system, replication and repair, as well as cell growth and death. The enrichment results also verified that the underlying mechanisms of FAA against breast cancer might be attributed to the coordinated regulation of several cancer-related pathways, such as the MAPK and mammalian target of rapamycin (mTOR) signaling pathways, among others. Conclusions: This study identified the potential targets and pathways of FAA in the treatment of breast cancer using a network pharmacology approach, and systematically elucidated the mechanisms of FAA in the treatment of breast cancer.
... Noscapine of Papaver somniferum is also reported to inhibit tumor growth and progression and interfering microtubules, has undergone phase I and phase II clinical trials (Henary et al, 2014;Chen et al, 2015). Artemisia absinthium, another novel source of phenolics compounds, flavonoids and essential oils with reported anti-proliferative and anti-apoptotic activity via Caspase 3 activation in HeLa cell lines (Kordali et al, 2005;Shafi et al, 2012). Chelidonium majus, the top natural cure for gallbladder stones was also reported to have cytotoxic effects on breast cancer cell lines (Aljuraisy et al, 2012). ...
... References used / extracts. Shafi et al (2012). absinthium) Parts usedaerial parts. ...
Full-text available
Plant extracts or plant products have been used from thousands of years all over the world in traditional medicine(TM) system includes Ayurveda, Unani, Siddha, Homeopathy, ancient Iran medicines, Islamic medicines, Chinese TM, Korean TM and African TM which are widely used for cancer prevention. They act as a biological response modifier or adaptogens, potentially enhancing the efficacy of the conventional therapies by interfering signaling pathways associated with cell growth and development.A range of clinical studies and research have done to identify the chemopreventive spectrum of the plants.However, some medicinal plants are also inherently toxic and various problems associated with them like quality issues, herb-drug interactions, bioavailability and lack of clinical trials which is essential for its safety and efficacy determination.The advent of nanomedicines has provided a way to deal with the poor bioavailability problems associated with herbal drugs.The approval of "PHY906" by USFDA for phase II clinical trial pave a way for future development of botanical drugs.
... Most of the studies are concentrated on A. absinthium, which have confirmed that A. absinthium extracts have an influence on the digestion system, due to their appetite-stimulating, antiulcer, and hepatoprotective effects, among others activities [13,19,[180][181][182][183][184]. Additionally, they have also shown, inter alia, cytotoxic, anthelmintic, antiprotozoal, analgesic, immunostimulating, cytotoxic, neuroprotective, and antidepressant activities [14][15][16][17][18]25,26,[30][31][32][33][34][35][36][37]86,122,130,[185][186][187][188][189][190][191]. ...
Full-text available
Artemisia species play a vital role in traditional and contemporary medicine. Among them, Artemisia abrotanum, Artemisia absinthium, Artemisia annua, Artemisia dracunculus, and Artemisia vulgaris are the most popular. The chemical composition and bioactivity of these species have been extensively studied. Studies on these species have confirmed their traditional applications and documented new pharmacological directions and their valuable and potential applications in cosmetology. Artemisia ssp. primarily contain sesquiterpenoid lactones, coumarins, flavonoids, and phenolic acids. Essential oils obtained from these species are of great biological importance. Extracts from Artemisia ssp. have been scientifically proven to exhibit, among others, hepatoprotective, neuroprotective, antidepressant, cytotoxic, and digestion-stimulating activities. In addition, their application in cosmetic products is currently the subject of several studies. Essential oils or extracts from different parts of Artemisia ssp. have been characterized by antibacterial, antifungal, and antioxidant activities. Products with Artemisia extracts, essential oils, or individual compounds can be used on skin, hair, and nails. Artemisia products are also used as ingredients in skincare cosmetics, such as creams, shampoos, essences, serums, masks, lotions, and tonics. This review focuses especially on elucidating the importance of the most popular/important species of the Artemisia genus in the cosmetic industry.
Full-text available
Introduction Artemisia absinthium (wormwood) exhibits anticancer properties by inhibiting proliferation and causing cell death in breast cancer. Targeted drug delivery of A. absinthium nanoformulation using N-isopropyl acrylamide, N-vinyl pyrrolidone, and acrylic acid-based polymeric nanoparticles (NVA-AA NPs) was ensured by utilizing features of the tumor microenvironment, although their mechanism of action involved in cytotoxicity remains unknown. Methods The present study employed nano LC-MS/MS to identify differences in secretory protein expression associated with the treatment of breast cancer cell lines (MCF-7; MDA-MB-231) by NVA-AA NPs for the determination of affected pathways and easily accessible therapeutic targets. Different bioinformatics tools were used to identify signature differentially expressed proteins (DEPs) using survival analysis by GENT2 and correlation analysis between their mRNA expressions and sensitivity toward small-molecule drugs as well as immune cell infiltration by GSCA. Results Analysis by GENT2 revealed 22 signature DEPs with the most significant change in their expression regulation, namely, gelsolin, alpha-fetoprotein, complement component C3, C7, histone H2B type 1-K, histone H2A.Z, H2AX, heat shock cognate 71 kDa protein, heat shock 70 kDa protein 1-like, cytochrome c somatic, GTP-binding nuclear protein Ran, tubulin beta chain, tubulin alpha-1B chain, tubulin alpha-1C chain, phosphoglycerate mutase 1, kininogen 1, carboxypeptidase N catalytic chain, fibulin-1, peroxiredoxins 4, lactate dehydrogenase C, SPARC, and SPARC-like protein 1. Correlation analysis between their mRNA expressions versus immune cell infiltrates showed a positive correlation with antitumor immune response elicited by these NPs as well as a correlation with drug response shown by the GDSC and CTRP drugs in different cancer cells. Discussion Our results suggest that NVA-AA NPs were able to invade the tumor microenvironment; transformed the communication network between the cancer cells; affected potential drivers of microtubular integrity, nucleosome assembly, and cell cycle; and eventually caused cell death.
Full-text available
Objective: A polyherbal medicine, Habb-e-Ustukhuddus (HU), is used for its anti-inflammatory properties. However, the anticancer and chemopreventive properties of HU were not known, and Therefore, investigated in the present study. Methods: Cancer cells were treated with 50-400 µg/ml HU and MTT, trypan blue, and clonogenic assays were performed. Propidium iodide (PI) staining, annexin V-FITC assay, and JC-1 staining were done for cell cycle progression, apoptosis, and mitochondrial membrane potential, respectively, using flow cytometry. Immunoblotting, cell migration and invasion assays were performed. Chemical characterization of HU was done through GC-MS and HPLC analyses. C57BL/6 mice were used to assess the in vivo toxicity of HU. Results: While evaluating the anticancer activity, the methanolic extract of HU (50-400 µg/ml) strongly inhibited the growth and survival (P<0.05-0.001) of lung and breast cancer cells and increased the cell population in the sub-G1 phase of the cell cycle. HU caused apoptotic death of cancer cells (P<0.05-0.001), which was associated with the depolarization of mitochondrial membrane potential (Δψ) (P<0.001) and an increase in Bax to Bcl-2 protein ratio. Further, HU inhibited the invasion and migration of cancer cells, which was accompanied by an increase in the epithelial marker, E-cadherin, and a decrease in the mesenchymal marker, vimentin. The HU characterization by GC-MS and HPLC analyses showed the abundance of bioactive compounds including flavonoids and alkaloids. In the chemopreventive study, the oral administration of methanolic extract of the formulation HU (50 and 100 mg/kg body weight) to mice did not cause any toxicity and significantly increased the specific activities of hepatic drug metabolizing phase I and phase II enzymes, which suggested for its detoxification potential of xenobiotic compounds. Conclusion: Together, these results demonstrated the anticancer potential HU, without any apparent toxicity in mice, and thus HU could be further explored for its clinical utility in cancer control.
Full-text available
Abstract Aim: This study was performed to evaluate the cytotoxic effects of hydroalcoholic extract of Artemisia sieberi and its effect on the cell cycle in the SKBr3 breast cancer cell line. Material and Methods: In this study, hydroalcoholic extract of Artemisia was prepared. SKBr3 cells were exposed to different concentrations (1, 10, 100, 1000 µgml-1) of extract at two different time intervals of 24 and 48 h. We employed MTT assay and flow cytometry analysis to evaluate effects of the prepared extract on the cell cycle characteristics and its cytotoxic properties. Results: Based on the data obtained from the MTT test, the highest toxicity of the extract observed at the concentration of 1000 µgml-1 within 48 h after the extract exposure. The IC50 of hydroalcoholic extract was 150 and 50 µgml-1 at 24 and 48 h, respectively. According to the data obtained from flow cytometry analysis, the extract arrests cell cycle in 24 and 48 h treatment groups. Conclusion: The hydroalcoholic extract of Artemisia at certain concentrations inhibits the growth of SKBr3 breast cancer cells by ceasing cell cycle step. Keywords: Artemisia, Breast cancer, Cell cycle, Hydroalcoholic extract
Objective(s): Increased quinolinic acid (QA) accumulation has been found in many neurodegenerative diseases. Artemisia absinthium (A. absinthium) has been reported to have neuroprotective and antioxidant activities. This study was designed to evaluate the effect of A. absinthium in QA-induced neurotoxicity in OLN-93 Cells. Methods: OLN-93 cells were cultured in a DMEM medium containing 10% (v/v) fetal bovine serum, 100 units/ml penicillin, and 100 μg/ml streptomycin. The cells were pretreated with concentrations of A. absinthium extract for two h and then exposed to QA for 24 h. After 24 h cell viability, the level of malondialdehyde (MDA), reactive oxygen species (ROS), and apoptotic cells were quantitated in OLN-93 Cells. Results: Pretreatment with A. absinthium extract prevented the loss of cell viability in OLN-93 cells. ROS generation, lipid peroxidation, and apoptosis in QA -injured OLN-93 cells were reduced following A. absinthium extract pretreatment. Conclusion: A. absinthium extract exerts its neuroprotective effect against QA-induced neurotoxicity via oxidative stress and apoptosis modulation.
Introduction. Recently, the anticancer activity of representatives of the genus Artemisia L. has been actively studied, and most studies are devoted to Artemisia annua L., which has been used since ancient times in the folk medicine of several countries as an antimalarial and anticancer agent. The similarity of the chemical composition predetermines the study of the anticancer activity of other species of the genus Artemisia L. The information about this is still not fully presented in scientific publications, is very diverse and sometimes even contradictory. Review of modern studies of anticancer activity of species of the genus Artemisia L., generalization of available data and providing information for future research is relevant. Text. The article presents a review of experimental data on the study of anticancer activity of representatives of the genus Artemisia L. It is noted that the main mechanism of such activity is apoptosis. Apoptosis is triggered by the increase of reactive oxygen species (ROS) inside cancer cells, reduction of mitochondrial membrane potential, activation of pro-apoptotic and, on the contrary, inhibition of anti-apoptotic proteins, as well as by formation of membrane bubbles, cell compression and by activation caspase. Conclusion. In the presented review, about 30 species of the genus Artemisia L. With the presented degree of study of this area, a number of questions remain unresolved. The most studied with respect to cytotoxic activity are Artemisia absinthium L. and Artemisia vulgaris L. In this aspect, the study of other closely related species of the genus Artemisia L. Also relevant is the study of cytotoxicity of representatives of the genus Artemisia L. on normal cell cultures and in comparison with positive control. In addition, a detailed study of the pool of secondary metabolites of different species of the genus Artemisia L. remains significant in order to reliably determine the components responsible for the manifestation of anticancer action. The pronounced effectiveness against cancer cells and, at the same time, a weak effect on healthy cells of the body of representatives of the genus Artemisia L. opens up the prospect of their use as sources of partner drugs with a synergistic effect and means of augmentation of antitumor therapy.
Artemisia absinthium leaves were utilized as a reducing agent for green synthesis of Zinc oxide nanoparticles (particle size 17 nm). Synthesized green-ZnO (g-ZnO) were characterized by SEM/EDX, FTIR, XRD, UV, and BET analyses and then further used as an adsorbent to remove Cr(VI) ions from simulated wastewater. Optimal pH, temperature and adsorbent dosage were determined through batch mode studies. High removal efficiency and adsorption capacity were observed at pH 4, 0.25 g L−1 dosage, and 25 mg L−1 concentration of Cr(VI). Experimental data were modelled with different adsorption kinetics (Elovich model, PFO, PSO, IDP model) and isotherms (Langmuir, Freundlich, and Temkin), and it was found the adsorption process was well fitted to Langmuir with an R2 value greater than>0.99. Computational calculation showed that the g-ZnO nanoparticles became ∼14 times more dynamic with delocalized surface states making them a relevant platform to adsorb Cr with greater work function compatibility supporting the experimental findings. The Qmax adsorption capacity of g-ZnO was 315.46 mg g−1 from Langmuir calculations. Thermodynamic calculations reveal that the Cr (VI) adsorption process was spontaneous and endothermic, with a positive ΔS value representing the disorder at the solid-solution interface during the adsorption. In addition, the present study has demonstrated that these g-ZnO nanoparticles show strong antibacterial activities against P. aeruginosa (MTCC 1688) and E. coli (MTCC 1687). Also, the novel g-ZnO adsorbent capacity to remove Cr(VI) from simulated water revealed that it could be reused at least six times with higher removal rates during regeneration experiments. The results obtained from adsorption and antimicrobial activities suggest that g-ZnO nanoparticles could be used effectively in real-time wastewater and agricultural safety applications.
Full-text available
Throughout medical history, plant products have been shown to be valuable sources of novel anti-cancer drugs. Examples are the Vinca alkaloids, the taxanes, and the camptothecins, derived from the Madagscan periwin-kle plant Catharantus roseus, the Pacific yew Taxus brevi-folia, and the Chinese tree Camptotheca acuminata, respectively. For this reason, the South-American Office for Anti-Cancer Drug Development has implemented a large-scale project of acquisition and testing of compounds isolated from South American medicinal plants. The species are selected on the basis of a potentially useful phytochemical composition by consulting ethnopharma-cological, chemosystemic, and ecological information. The collected samples are dried and first extracted with an organic solvent, then with distilled water. These crude extracts are evaluated at a concentration of 50 µg/ml for antiproliferative activity against one cell line. Extracts that significantly inhibit the growth of the cells (≥50%) at relatively low concentrations (≤50 µg/ml) are submitted to the more comprehensive disease-oriented screen of the U.S. National Cancer Institute. In parallel, these samples are further purified by bioassay-guided purification, involving repeated fractionation by diverse chromatography methods. If the active substance is expected to represent a novel structure, it is identified by appropriate chemical techniques, mechanistic studies are performed with a wide diversity of tumor models and laboratory techniques, and efforts are undertaken for the synthesis of potentially more useful analogs.
Full-text available
Cancer remains one of the most dreaded diseases causing an astonishingly high death rate, second only to cardiac arrest. The fact that conventional and newly emerging treatment procedures like chemotherapy, catalytic therapy, photodynamic therapy and radiotherapy have not succeeded in reverting the outcome of the disease to any drastic extent, has made researchers investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. This study progresses in the direction of identifying component(s) from Nigella sativa with anti cancer activity. In the present study we investigated the efficacy of Organic extracts of Nigella sativa seed powder for its clonogenic inhibition and induction of apoptosis in HeLa cancer cell. Methanolic, n-Hexane and chloroform extracts of Nigella sativa seedz effectively killed HeLa cells. The IC50 values of methanolic, n-hexane, and chloroform extracts of Nigella sativa were 2.28 microg/ml, 2.20 microg/ml and 0.41 ng/ml, respectively. All three extracts induced apoptosis in HeLa cells. Apoptosis was confirmed by DNA fragmentation, western blot and terminal transferase-mediated dUTP-digoxigenin-end labeling (TUNEL) assay. Western Blot and TUNEL results suggested that Nigella sativa seed extracts regulated the expression of pro- and anti- apoptotic genes, indicating its possible development as a potential therapeutic agent for cervical cancer upon further investigation.
Ve have used a structure-activity approach to investigate whether the Vinca alkaloids inhibit cell proliferation primarily by means of their effects on mitotic spindle microtubules or by another mechanism or by a combination of mechanisms. Five Vìnca alkaloids were used to investigate the relationship in Urla cells between inhibition of cell proliferation and blockage of mitosis, alteration of spindle organization, and depolymeri- zation of microtubules. Indirect immunofluorescence staining of micro- tubules and 4,6-diamidino-2-phenylindole staining of chromatin were used to characterize the effects of the drugs on the distributions of cells in stages of the cell cycle and on the organization of microtubules and chromosomes in metaphase spindles. The microtubule polymer was iso lated from cells and quantified using a competitive enzyme-linked im- munoadsorbent assay for tubulin. We observed a nearly perfect coinci dence between the concentration of each Vinca derivative that inhibited cell proliferation and the concentration that caused 50% accumulation of cells at metaphase, despite the fact that the antiproliferative potencies of the drugs varied over a broad concentration range. Inhibition of cell proliferation and blockage of cells at metaphase at the lowest effective concentrations of all Vinca derivatives occurred with little or no micro- tubule depolymerization or spindle disorganization. With increasing drug concentrations, the organization of microtubules and chromosomes in arrested mitotic spindles deteriorated in a manner that was common to all five congeners. These results indicate that the antiproliferative activity of the Vinca alkaloids at their lowest effective concentrations in Ik-la cells is due to inhibition of mitotic spindle function. The results suggest further that the Vinca alkaloids inhibit cell proliferation by altering the dynamics of tubulin addition and loss at the ends of mitotic spindle microtubules rather than by depolymerizing the microtubules. The spe cific alterations of spindle microtubule dynamics appear to differ among the five Vinca congeners, and such differences may be responsible for differences in the antitumor specificities of the drugs.
Epidemiological studies have described the beneficial effects of dietary polyphenols (flavonoids) on the reduction of the risk of chronic diseases, including cancer. Moreover, it has been shown that flavonoids, such as quercetin in apples, epigallocatechin-3-gallate in green tea and genistein in soya, induce apoptosis. This programmed cell death plays a critical role in physiological functions, but there is underlying dysregulation of apoptosis in numerous pathological situations such as Parkinson's disease, Alzheimer's disease and cancer. At the molecular level, flavonoids have been reported to modulate a number of key elements in cellular signal transduction pathways linked to the apoptotic process (caspases and bcl-2 genes), but that regulation and induction of apoptosis are unclear. The aim of this review is to provide insights into the molecular basis of the potential chemopreventive activities of representative flavonoids, with emphasis on their ability to control intracellular signaling cascades responsible for regulating apoptosis, a relevant target in cancer-preventive approach.
We investigated mechanism(s) where propolis induces apoptosis in human leukemic U937 cells. Propolis inhibited the proliferation of U937 cells in a dose-dependent manner by inducing apoptosis and blocking cell cycle progression in the G2/M phase. Western blot analysis showed that propolis increases the expression of p21 and p27 proteins, and decreases the levels of cyclin B1, cyclin A, Cdk2 and Cdc2, thereby contributing to cell cycle arrest. DAPI staining assay revealed typical morphology features of apoptotic cells. Propolis-induced apoptosis was also confirmed by assays with annexin V-FITC, PI-labeling and DNA fragmentation assay. The increase in apoptosis level induced by propolis was associated with down-regulation of Bcl-2 and activation of caspase-3, but not with Bax. These results suggests that propolis-induced apoptosis is related to the selective activation of caspase-3 and induction of Bcl-2/Bax regulation.
Escape from apoptosis is often a hallmark of cancer cells, and is associated to chemotherapy resistance or tumor relapse. Proteins from the Bcl-2 family are the key regulators of the intrinsic pathway of apoptosis, controlling the point-of no-return and setting the threshold to engage the death machinery in response to a chemical damage. Therefore, Bcl-2 proteins have emerged as an attractive target to develop novel anticancer drugs. Current pharmacological approaches are focused on the use of peptides, small inhibitory molecules or antisense oligonucleotides to neutralize antiapoptotic Bcl-2 proteins, lowering the threshold and facilitating apoptosis of cancer cells. We discuss here recent advances in the development of Bcl-2 targeted anticancer therapies.