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Huaier Regulates Cell Fate by the Rescue of Disrupted Transcription Control in the Hippo Signaling Pathway
... Currently we are launching into the postpandemic COVID-19 era and have found that more problems remain to be solved, such as more efficient preventive strategies and/or complications after invasive treatments. We have started genomic and genetic analysis of mRNA vaccination to our cancer patients from April 2020 [1,[8][9][10][11][12][13][14][15], and examined any alterations in genomic stability and in subsequent translation and transcription processes [12]. Our previous research results demonstrated that extensive genomic flexibility and capability were derived from opportunistic virus infections, and that those genomic potential is the basis to recover from the diseases [12]. ...
... Our previous research results demonstrated that extensive genomic flexibility and capability were derived from opportunistic virus infections, and that those genomic potential is the basis to recover from the diseases [12]. Huaier (Trametes robiniophila murr), natural herb therapy on cancer patients (Chinese Administrative License No. Z-20000109) [8][9][10][11][12][13][14][15], contributes to making this genomic potential efficient for the rescue of disrupted physiological signaling networks. We found that Huaier could compensate for any damage from destructive ribosomal RNA structures after mRNA vaccination against SARS-CoV-2, dependent on the genomic potential of each individual, independent of the severity of cancer or basic health conditions [1]. ...
... A small dose of Huaier, 6 g per day, is enough to prevent SARS-CoV-2 infection as well as any unexpected damage in molecular systems in this normal control. Huaier effects contributed to control kinase function regulation via mTOR/PI3K/AKT signaling networks, cooperated with massive mi-and pi-RNAmedicated transcriptional control [1,[8][9][10][11][12][13][14][15]. The present study thus provides an efficient and non-hazardous strategy to cope with the up-coming postpandemic COVID-19 era. ...
... Although medical doctors usually pursuit and monitor the results of clinical blood tests and CT or MRI image analysis, but the individual recognition of recovery process in daily life has as much impact. Huaier effects on tissue regeneration and repair has been emphasized in the previous reports through a series of our clinical research [1][2][3][4][5][6][7][8][9][10][11][12] , which seemed to contribute more than the effects on cancer cellspeci c cell death. The inhibition of cancer progression observed in Huaier therapy depends on the transcriptional regulation of pluripotency of stem cell 1,[4][5][6][7][8][9][10][11] . ...
... Through our clinical research from 2018, we have reported sequentially about the molecular basis of anticancer effects of Huaier, and that the results on the minimum essential rescue of disruption and dysfunctions in the integrated signaling pathway networks by miRNA-mediated transcriptional control [1][2][3][4][5][6][7][8][9][10][11][12] . Hair growth and restoration is a typical proof of tissue regeneration. ...
... The present study successfully identi ed the Huaier effects on regulating Hedgehog signaling pathway functions 22, 23 within 1-3 months after administration, and that in a dose dependent manner. Huaier administration initiated massive hair growth with an increase of body weights t over 5 Kg /month, with healthy appetite, and totally cure of skin lesions as shown in the previous report 12 . Although Huaier has an effect to rewind the life time (anti-ageing) [1][2][3][4][5][6][7][8][9][10][11][12] , as an effect as (tyrosine) kinase regulator 2,8 , it is noteworthy to observe the hair restoration with black hair (typical for Japanese population) even over the age of 70's. ...
In the process of caner recovery, Huaier simultaneously promotes significant hair growth and restoration as a typical evidence of its efficacy on induced tissue regeneration and repair function. Hair growth after conventional chemotherapy was a first and significant observation, and that in dose-dependent manner. The molecular basis of hair growth was identified as the activation of Hedgehog signaling pathway and among various perturbed signal transfer network, which initiates systematic tissue repair not only in pathogenic lesions but also to skin problems and inter/intra neural signal transfer. The activation followed by the inhibition at 3-6 months interval resulted in the prevention of subsequent complications, such as recurrence and metastasis by transcriptional control in cancer stem cell production. This regulation was demonstrated to occur irrespectively to SMO (smoothhead) function. The present study thus provides a simple and clear proof of Huaier effects on tissue regeneration, additional to the reported cancer specific-cell death.
... We ourselves have confirmed the drastic improvements not only in cancer patients, but also in the other disorders in skin, liver, kidney diseases and disorders, and more importantly, neurodegenerative diseases such as Parkinson's disease [16]. In China, the basic investigations revealed the effects by in vitro experiments and using both mice and rats inoculated with cancer cells, in which. ...
... Not only in cancer cases, we observed multi-dimensional efficacies of Huaier, especially on liver dysfunction, nephritis, bronchiectasis, constipation and skin lesions in a dose-dependent manner [16]. ...
... We made a hypothesis that Huaier efficacy should be related to the multi-dimensional oncogenic potential such as Hippo signaling pathway [21,22], and proved this hypothesis by a simple experiment using mutant Drosophila strain with disrupted Hippo signaling pathway. The Huaier treatment successfully rescued disrupted function in the mutant fly, with a dose-dependent manner, just the same as shown in cancer patients [16]. This rescue of Hippo signaling pathway function was derived from the rescue in disrupted transcriptional control, and accompanied with a strong shift of the metabolome profile to the early embryonic pattern to "rejuvenescence" form, as predicted in B. C. 220. ...
Preface Cell is full of signals and networks, which link intensively each other. In Figure 1, the symbolic network, intra-cellular multi-signal trafficking on metabolic pathways were shown with a comparison to integrated circuit (IC) chart. These panels showed strong similarity between integrated signal traffics in the cell and the superconductor substrate.
... At the beginning of our investigation on molecular basis of anti-cancer efficacy of Huaier, we used transgenic Drosophila flies with overexpressing nonphosphorylatable Yorkie (Yki:V5S168A) as a cancer model [3]. This model was later revealed to be useful to screen the candidate compound for amyotrophic lateral sclerosis (ALS), however, nearly 5 years' research resulted in vain. ...
... Total RNA-and small non-coding RNA-sequencing on the MGISEQ-2000 and BGISEQ-500 platform [26,27] were processed in BGI, Shenzhen, China, as descrived previously [4][5][6][7][8][9][10]. The subseuent bioinformatics work was also processed in BGI, [3,4,9,28]. ...
... The post-transcriptional control, epigenetic processes seemd to matter the differences in pathogenecity among family members, from the observation of the altered levels of DNA repair enzymes such as MGMT, MLH1-3, and p53 [5,7]. In addition, miRNA genes are also drastically altered which associated with CpG islands, that may be repressed by epigenetic methylation [3,7]. were detected to support the compensation of defected functions in signaling pathways such as; ...
... Here we report the further analysis of Huaier efficacy on the prevention of cancer progression and on the tissue regeneration with normal differentiated cells. We have already reported the significant impact of Huaier on the transcriptional control mediated by microRNA and small non-coding RNAs, and the potential for tissue regeneration by the rescue of Hippo signaling pathway [13,14]. In the present study, we provide further insights into the none the less essential effects of Huaier for promoting cancer recovery by utilizing stem cell production and normal differentiation. ...
... For the repair function, there should be enough cells to regenerate, including the potential of cancer stem cells for normal differentiation. We have already indicated the potential of Huaier to determine the cell fate to normal differentiation by the rescue of Hippo signaling pathway functions [13] and here we present more detailed information related to tissue regeneration in the process of Huaier treatment. ...
... Huaier compounds were provided by the manufacturer for this purpose with a strict control on transfer to Japan, good condition for maintenance, and provision to the patient volunteers, just as the same as the previous reports [3][4][5][6]13]. ...
The induced normal tissue regeneration was the results of transcriptional control of iPS/ES cell production. Among the gene families to control iPS/ES production, especially c-my expression level played a major role among the other genes or gene families. These results provide a clue to clarify the Huaier effects not only for recovery from cancer, but also for the prevention of many related diseases and disorders caused by daily accumulation of environmental stresses and ageing by controlling normal tissue regeneration by stem cell control.
... After we succeeded to prove the molecular basis of anti-cancer effects of Huaier by a simple method using Drosophila mutants [4], we initiated clinical research total RNA and small non-coding RNA analysis to confirm the reality of significance of Huaier treatment on cancer [5][6][7]. The results provided the massive alteration and modification of the transcripts by upand down-regulation, based on the miRNA-mediated transcriptional control [5][6][7]. ...
... The identification of EGFR as an oncogene has led to the series of development of highly toxic anticancer therapeutics directed against EGFR (called "EGFR inhibitors", EGFRi), including recent development of Osimertinib, a third-generation tyrosine kinase inhibitor[27]. Huaier treatment was simultaneously proved its significant efficacy on breast cancer, chiefly by the rescue of defected function in the transcriptional control of Hippo signaling pathway[4,28,29]. ...
... Data on molecular level of each patient and controlling agents are strongly lacking to determine the combination of chemotherapeutic agents for their efficacy and safety. Huaier (Trametes robiniophila murr) has been introduced its significant anti-cancer efficacy, and more importantly, without any adverse events and/or toxicity [3][4][5][6][7][8]. We have proved the molecular basis for Huaier effects based on the rescue of disrupted Hippo signalling pathway in Drosophila mutants, especially through the rescue of transcriptional dysregulation [5,9]. ...
... The results provided estimated transcribed genes of >27,000; mean number of 27, 447 per sample, and the average mapping ratio with reference genome is 89.64 % (varies from 71.49% to 94.51%) as previously reported. The obtained additional novel genes and small nuclear non-coding RNA obtained in the present study have been uploaded to the genes already deposited to the NCBI GEO (GSE157086)[6,7]. ...
:We have initiated genome-scope project for precise comprehension of anti-cancer effects of Huaier, with a comparison to the results by the treatment with platinum (II) complex (FOLFOX6, FORLFIRINOX, and Cisplatin:CDDP).
... In the course of our clinical research to investigate molecular basis of anti-cancer efficacy of Huaier [6][7][8][9][10][11][12][13][14][15][16], we happened to observe several patients suddenly deteriorated without any considerable factors. The patients with Huaier therapy maintained good quality of life (QOL) for more than expected by any means, and confirmed the recovery judged by any medical tests including CT and MRI images. ...
Although striking effects of vaccination strategy against COVID-19 world-wide, a
long-term influence by sequential viral mRNA injections are unknown. We analysed
biological alterations by total RNA sequencing in Pfizer-BioNTech vaccinated normal
healthy volunteers and cancer patients, with or without adjuvant Huaier therapy.
A significant destruction in ribosomal RNA structures was identified, enhanced
by serial shots. Unlike the destruction caused by chemotherapy with platinum
(II) complex, progressive destruction in 18S ribosome was identified even at
6 months after vaccination. The influence resulted in massive inhibition of translation
and transcription, significantly in intra/inter neural signaling transfer and in lipid
metabolism, related to ageing process. Huaier compensated these dysfunctions by
miRNA-mediated transcriptional control, by typical activation in PI3K/AKT signaling
pathway. Gene Ontology analysis revealed spontaneous virion production in number
even after 3 months of the first vaccination. Present study indicated that the adjuvant
therapy like Huaier compensates accelerated ageing process by mRNA vaccination.
... We have reported with simple and successful evidence that the rescue of Hippo signaling pathway was a molecular basis of anticancer effects of Huaier [14] in 2017. Thereafter many researches followed for the confirmation, to provide the evidence in vitro that deregulation in Hippo signaling pathway, disruption of transcription control is the major course of carcinogenesis [15][16][17][18][19][20][21][22]. ...
Prostate cancer is classified as mild malignant tumor, since the
growth is slow, and also the early detection and monitoring of PSA
and other cancer cell-specific markers are available among the target
population in walk-in clinic even effect the bones and lymph nodes,
it requires long term treatments which significantly decrease the
lifespan in patients. Trametes robiniophila Murr (Huaier) has proved
broad spectrum anti-cancer effects, which initiates the recovery of
damaged bio physiological functions in the end by dose-dependent
manner which proved the molecular basis of Huaier effects by
total RNA and small non-coding RNA sequencing (“genomescope”
project) and that is based on the rescue of the disrupted
transcriptional control based on individual capability and flexibility of
genomic potential. Here we focused to show MEGA-DATA genome
analysis results on inhibition of active progression in cancer in situ, and also prevents relapse of prostate cancer. Total sequencing
of RNAs revealed massive SNP variances (average 89,473SNP
variances per individual), however, it is unlikely that specific type
variants influenced to the malignancy, process, and prognosis.
On the other hand, the significant up-regulation and alteration of
major ontogenesis and tumor suppressor genes were detected in
transcribed genes, especially in altered (normalized) NFκB, TGFb,
BRCA2 and p53 genes, and their leading signaling pathways.
These genetic alterations in transcriptomes were based on miRNAmediated
transcriptional control as reported drugs have not shown
any effect. Thus, the present study provides the safe and effective
treatment to prevent and inhibit prostate cancer progression, and
also to maintain homeostasis in a long-range of stressful human life
without excessive medical treatments.
... Similarly, there are some novel mutations reported by other countries and researchers, such as c.457G > A (p.E153K) from Iran [96]; compound heterozygous genotype p.Y140C/p.R297X from Canada [97]; c.1705C > A (p.Q569K) novel and c.1562C > T/c.1705C > A heterozygote mutations from China [98]; and K255Rfs*18, E153Rfs*39, and Q350* from Egypt [91]. ...
Mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders caused by a lysosomal enzyme deficiency or malfunction, which leads to the accumulation of glycosaminoglycans in tissues and organs. If not treated at an early stage, patients have various health problems, affecting their quality of life and life-span. Two therapeutic options for MPS are widely used in practice: enzyme replacement therapy and hematopoietic stem cell transplantation. However, early diagnosis of MPS is crucial, as treatment may be too late to reverse or ameliorate the disease progress. It has been noted that the prevalence of MPS and each subtype varies based on geographic regions and/or ethnic background. Each type of MPS is caused by a wide range of the mutational spectrum, mainly missense mutations. Some mutations were derived from the common founder effect. In the previous study, Khan et al. 2018 have reported the epidemiology of MPS from 22 countries and 16 regions. In this study, we aimed to update the prevalence of MPS across the world. We have collected and investigated 189 publications related to the prevalence of MPS via PubMed as of December 2020. In total, data from 33 countries and 23 regions were compiled and analyzed. Saudi Arabia provided the highest frequency of overall MPS because of regional or consanguineous marriages (or founder effect), followed by Portugal, Brazil, the Netherlands, and Australia. The newborn screening is an efficient and early diagnosis for MPS. MPS I has been approved for newborn screening in the United States. After the newborn screening of MPS I, the frequency of MPS I increased, compared with the past incidence rates. Overall, we conclude that the current identification methods are not enough to recognize all MPS patients, leading to an inaccurate incidence and status. Differences in ethnic background and/or founder effects impact on the frequency of MPS, which affects the prevalence of MPS. Two-tier newborn screening has accelerated early recognition of MPS I, providing an accurate incidence of patients.
... Similarly, there are some novel mutations reported by other countries and researchers, such as c.457G > A (p.E153K) from Iran [96]; compound heterozygous genotype p.Y140C/p.R297X from Canada [97]; c.1705C > A (p.Q569K) novel and c.1562C > T/c.1705C > A heterozygote mutations from China [98]; and K255Rfs*18, E153Rfs*39, and Q350* from Egypt [91]. ...
Mucopolysaccharidoses (MPS) are a group of lysosomal storage disorders caused by a lysosomal enzyme deficiency or malfunction, which leads to the accumulation of glycosamino-glycans in tissues and organs. If not treated at an early stage, patients have various health problems , affecting their quality of life and lifespan. Two therapeutic options for MPS are widely used in practice: enzyme replacement therapy and hematopoietic stem cell transplantation. However, early diagnosis of MPS is crucial, as treatment may be too late to reverse or ameliorate the disease progress. It has been noted that the prevalence of MPS and each subtype varies based on geographic regions and/or ethnic background. Each type of MPS is caused by a wide range of the mutational spectrum, mainly missense mutations. Some mutations were derived from the common founder effect. In the previous study, Khan et al. 2018 have reported the epidemiology of MPS from 22 countries and 16 regions. In this study, we aimed to update the prevalence of MPS across the world. We have collected and investigated 189 publications related to the prevalence of MPS via PubMed as of December 2020. In total, data from 33 countries and 23 regions were compiled and analyzed. Saudi Arabia provided the highest frequency of overall MPS because of regional or consanguineous marriages (or founder effect), followed by Portugal, Brazil, the Netherlands, and Australia. The newborn screening is an efficient and early diagnosis for MPS. MPS I has been approved for newborn screening in the United States. After the newborn screening of MPS I, the frequency of MPS I increased, compared with the past incidence rates. Overall, we conclude that the current identification methods are not enough to recognize all MPS patients, leading to an inaccurate incidence and status. Differences in ethnic background and/or founder effects impact on the frequency of MPS, which affects the prevalence of MPS. Two-tier newborn screening has accelerated early recognition of MPS I, providing an accurate incidence of patients.
... Total 7,664 DEGs coding transcriptional factors were altered to rescue and/or modify transcriptional misregulation noted in Figure 3b, within 30 days after Huaier administration. We have already indicated Huaier effects on the rescue of transcription control using Drosophila mutants [3], and the present study provides the systematic and quantitative background. These dynamic changes in transcription factors might explain the enormous range of transcription control rescue by Huaier. ...
... The sampling materials were total blood, the same as reported previously7,8 , by the consideration of as follows; 1) easy to obtain from any candidate patients, 2) non-or less-hazardous procedure, and 3) applicable to any patients of any stages, even after the surgical operation. To compare with the other sampling, RNA extraction using nuclear cell components in peripheral blood rapidly reflects the biophysiological changes, and that more sensitively5,6 to monitor the course of any treatment than any other samples such as dissected organs.Patient characteristics and sample collection. Basically, the volunteers were chosen on the demand of each individual, with a condition as written below; 1) diagnosed as cancer in the clinics, 2) exclusive for anti-cancer chemotherapy, radiotherapy, or any measures to influence genomic or genetic events in physiological condition for 90 days research period, 3) between ages of 35 to 85. ...
Clinical significance of anti-cancer effects of Huaier has been emphasized recently. We have proved that a broad spectrum of Huaier effects was based on the rescue of the disrupted Hippo signalling pathway, especially through the rescue of transcriptional dysregulation. We initiated clinical research for thorough understanding of mode of action by total RNA- and small non-coding RNA-sequencing. Here we provide the surprising genomic plasticity by Huaier observed in the time course of recovery from cancer. The extensive changes in RNA editing events were first observed, such as average 92,427 SNP variances per individual (22,688 in normal control). The subsequent changes in the process of translation and transcription resulted in the drastic changes in number of up/down-regulated transcripts. The ratio of the changes at maximum was 85% (23,210/27,447). With the advent of many novel sequences found in small nuclear RNAs and the consequent gene-silencing derived from miRNA-mediated post-transcriptional control, quantitative and qualitative changes of expression in transcriptional factors (mean 1,115/person) contributed to rescue dysregulated functions occurred in almost all required physiological functions for cancer recovery. The transcription control was demonstrated as functional lineage map between the core transcriptional factor and functionally linked transcripts, and the recovery could be clearly shown as the silencing of those massive changes in the end. The genetic alterations, especially related to the inhibition of NFκB and TGF-β signalling pathways showed significant effects on the improvement of prognosis. Thus, Huaier therapy contributes to cancer cell death with damaged tissue regeneration by extensive and systematic genomic modulations.
Breast cancer is one of the most common female malignant tumors today and represents a serious health risk for women. Although the survival rate and quality of life of patients with breast cancer are improving with the continuous development of medical technology, metastasis, recurrence, and drug resistance of breast cancer remain a significant problem. Huaier, a traditional Chinese medicine (TCM) fungus, is a type of Sophora embolism fungus growing on old Sophora stems. The polysaccharides of Trametes robiniophila Murr (PS-T) are the main active ingredient of Huaier. There is increasing evidence that Huaier has great potential in breast cancer treatment, and its anti-cancer mechanism may be related to a variety of biological activities, such as the inhibition of cell proliferation, metastasis, tumor angiogenesis, the promotion of cancer cell death, and regulation of tumor-specific immunity. There is growing evidence that Huaier may be effective in the clinical treatment of breast cancer. This review systematically summarizes the basic and clinical studies on the use of Huaier in the treatment of breast cancer, providing useful information to guide the clinical application of Huaier and future clinical studies.
Although striking effects of vaccination strategy against COVID-19 world-wide, a long-term influence by sequential viral mRNA injections are unknown. We analysed biological alterations by total RNA sequencing in Pfizer-BioNTech vaccinated normal healthy volunteers and cancer patients, with or without adjuvant Huaier therapy. A significant destruction in ribosomal RNA structures was identified, enhanced by serial shots. Unlike the destruction caused by chemotherapy with platinum (II) complex, progressive destruction in 18S ribosome was identified even at 6 months after vaccination. The influence resulted in massive inhibition of translation and transcription, significantly in intra/inter neural signaling transfer and in lipid metabolism, related to ageing process. Huaier compensated these dysfunctions by miRNA-mediated transcriptional control, by typical activation in PI3K/AKT signaling pathway. Gene Ontology analysis revealed spontaneous virion production in number even after 3 months of the first vaccination. Present study indicated that the adjuvant therapy like Huaier compensates accelerated ageing process by mRNA vaccination.
Huntington's disease (HD) is a progressive, dominantly inherited neurological disorder caused by an abnormal expansion of polyglutamine (polyQ) repeat within the Huntingtin (Htt) protein with no disease modifying treatments. In a Drosophila model of HD, expression of mutant Huntingtin (Htt) protein with expanded polyQ leads to formation of inclusion bodies (IBs), increase in cellular toxicity, progression of motor disabilities and reduced viability. Multiple cellular events such as oxidative stress, mitochondrial dysfunction, inflammation and transcriptional dysregulation are reported to contribute to pathology, however, till date there are no disease-modifying treatments with least side effects. Therefore, we investigated effect of the phytochemical curcumin on HD pathogenesis. Curcumin, a phytochemical and commonly used ingredient in Asian food has a wide spectrum of anti-oxidant, anti-inflammatory and anti-fibrilogenic properties. In this study, we provide evidence that curcumin significantly ameliorates disease symptoms in a Drosophila model of HD by suppressing cell death and can be a key to halting the progression of Huntington's disease with least side effects.
Huaier extract is attracting increased attention due to its biological activities, including antitumor, anti-parasite and immunomodulatory effects. Here, we investigated the role of autophagy in Huaier-induced cytotoxicity in MDA-MB-231, MDA-MB-468 and MCF7 breast cancer cells. Huaier treatment inhibited cell viability in all three cell lines and induced various large membranous vacuoles in the cytoplasm. In addition, electron microscopy, MDC staining, accumulated expression of autophagy markers and flow cytometry revealed that Huaier extract triggered autophagy. Inhibition of autophagy attenuated Huaier-induced cell death. Furthermore, Huaier extract inhibited the mammalian target of the rapamycin (mTOR)/S6K pathway in breast cancer cells. After implanting MDA-MB-231 cells subcutaneously into the right flank of BALB/c nu/nu mice, Huaier extract induced autophagy and effectively inhibited xenograft tumor growth. This study is the first to show that Huaier-induced cytotoxicity is partially mediated through autophagic cell death in breast cancer cells through suppression of the mTOR/S6K pathway.
In recent years, aqueous extract of Trametes robiniophila Murr. (Huaier), a traditional Chinese medicine, has been frequently used in China for complementary cancer therapy. However, the mechanisms underlying its anticancer effects have yet to be elucidated. The present study aimed to evaluate the ability of Huaier extract to inhibit proliferation, promote apoptosis and suppress mobility in the fibrosarcoma HT1080 cell line in vitro. The cells were treated with gradient doses of Huaier extract at concentrations of 0, 4, 8 or 16 mg/ml for 24, 48 or 72 h. The cell viability and motility were measured in vitro using MTT, invasive, migration and scratch assays. The distribution of the cell cycle and the extent of cellular apoptosis were analyzed by flow cytometry. The apoptotic pathways were detected using a mitochondrial membrane potential transition assay and western blotting. The results revealed that the cellular viability decreased significantly with increasing concentrations of Huaier extract. In addition, cell invasiveness and migration were also suppressed significantly. It was demonstrated that Huaier extract induced G2 cell-cycle arrest and cellular apoptosis in a time- and dose-dependent manner. The decreased mitochondrial membrane potential, the downregulation of B-cell lymphoma 2 and pro-caspase-3, and upregulation of Bcl-2-associated X protein, cleaved caspase-9 and caspase-3 suggested that Huaier extract induced the apoptosis of HT1080 cells through the mitochondrial pathway. The results of the present study indicate that Huaier extract is a potential complementary agent for the treatment of fibrosarcoma.
The DRE/DREF transcriptional regulatory system has been demonstrated to activate a wide variety of genes with various functions. In Drosophila, the Hippo pathway is known to suppress cell proliferation by inducing apoptosis and cell cycle arrest through inactivation of Yorkie, a transcription co-activator. In the present study, we found that half dose reduction of the hippo (hpo) gene induces ectopic DNA synthesis in eye discs that is suppressed by overexpression of DREF. Half reduction of the hpo gene dose reduced apoptosis in DREF-overexpressing flies. Consistent with these observations, overexpression of DREF increased the levels of hpo and phosphorylated Yorkie in eye discs. Interestingly, the diap1-lacZ reporter was seen to be significantly decreased by overexpression of DREF. Luciferase reporter assays in cultured S2 cells revealed that one of two DREs identified in the hpo gene promoter region was responsible for promoter activity in S2 cells. Furthermore, endogenous hpo mRNA was reduced in DREF knockdown S2 cells, and chromatin immnunoprecipitation assays with anti-DREF antibodies proved that DREF binds specifically to the hpo gene promoter region containing DREs in vivo. Together, these results indicate that the DRE/DREF pathway is required for transcriptional activation of the hpo gene to positively control Hippo pathways.
The Drosophila melanogaster embryo has been widely utilized as a model for genetics and developmental biology due to its small size, short generation time, and large brood size. Information on embryonic metabolism during developmental progression is important for further understanding the mechanisms of Drosophila embryogenesis. Therefore, the aim of this study is to assess the changes in embryos' metabolome that occur at different stages of the Drosophila embryonic development. Time course samples of Drosophila embryos were subjected to GC/MS-based metabolome analysis for profiling of low molecular weight hydrophilic metabolites, including sugars, amino acids, and organic acids. The results showed that the metabolic profiles of Drosophila embryo varied during the course of development and there was a strong correlation between the metabolome and different embryonic stages. Using the metabolome information, we were able to establish a prediction model for developmental stages of embryos starting from their high-resolution quantitative metabolite composition. Among the important metabolites revealed from our model, we suggest that different amino acids appear to play distinct roles in different developmental stages and an appropriate balance in trehalose-glucose ratio is crucial to supply the carbohydrate source for the development of Drosophila embryo.
The theory of targeting cancer stem-like cells (CSCs) provides novel strategy for cancer treatment. In the present study, we examined the inhibitory effect of Huaier aqueous extract on eradicating breast cancer stem cells and explored the underlying mechanisms. Our data demonstrated that various concentrations of Huaier extract significantly decreased the viabilities, numbers, and sizes of mammospheres. After incubation with Huaier extract for 24 h, the clonogenicity of MCF7 cell line was obviously impaired, along with less holoclones. In addition, Huaier extract reduced the number of cells expressing CD44+/CD24- and decreased the level of stem cell markers (OCT-4, NESTIN, and NANOG). The hedgehog (Hh), notch, and Wnt/β-catenin pathways were essential stem cell signaling pathways involved in regulating CSC renewal and maintenance. We reported that the inhibitory effect of Huaier extract was partly depended on the inactivation of Hh pathway. These findings provided experimental evidence that Huaier extract was a promising therapeutic drug for eliminating the breast cancer stem cells.
Various studies have reported that Huaier possesses anti-tumor effects. However, the mechanisms are not completely elucidated. Here, we found 66 differentially expressed miRNAs in Huaier-treated pulmonary adenocarcinoma A549 cells, with upregulation of miR-26b-5p. Transfection of A549 cells with miR-26b-5p mimic inhibited proliferation and induced apoptosis, while transfection of Huaier-treated A549 cells with a miR-26b-5p inhibitor reversed the effects of Huaier. EZH2 was verified as the target of miR-26b-5p. Thus, our findings indicate that Huaier might suppress proliferation and induce apoptosis in lung cancer cells via a miR-26b-5p-EZH2-mediated approach, which provides a new perspective for understanding the anti-tumor effects of Huaier.
Traditional Chinese medicine has gained popularity due to its ability to kill tumor cells. Recently, the apoptotic and anti-angiogenic effects of Trametes robiniophila murr (Huaier) have been investigated. The aim of this study was to investigate its effect on cell mobility and tumor growth in ovarian cancer. Cell viability and motility were measured using SRB, scratch and migration assays. Cell apoptosis was analysed by annexin V/PI staining. Using a reverse-phase protein array (RPPA) assay, we analyzed the levels of 153 proteins and/or phosphorylations in Huaier-treated and untreated cells. Huaier inhibited cell viability and induced both early and late apoptosis in SKOV3, SKOV3.ip1 and Hey cells in a time- and dose-dependent manner. Cell invasiveness and migration were also suppressed significantly. The RPPA results showed significant differences (of at least 30%; P <0.05) in the levels of 7 molecules in SKOV3 cells and 10 in SKOV3.ip1 cells between the untreated and treated cells. Most of the molecules identified play roles in cell proliferation, apoptosis or cell adhesion/invasion. Western blot analysis further validated that Huaier treatment resulted in decreased AKT phosphorylation, enhanced expression of total GSK3β, inhibition of the phosphorylation of GSK3β on S9, reduction of both cytoplasmic β-catenin expression and nuclear β-catenin translocation, and transcriptional repression of several Wnt/β-catenin target genes (DIXDC1, LRP6, WNT5A, and cyclin D1). After knocking down GSK3β, β-catenin expression could not be inhibited by Huaier. Finally, Huaier inhibited the growth of ovarian tumor xenografts in vivo. These studies indicate that Huaier inhibits tumor cell mobility in ovarian cancer via the AKT/GSK3β/β-catenin signaling pathway.
The existence of cancer stem cells (CSCs) is central to the pathogenesis and therapy resistance of colorectal cancer. The aim of this study was to evaluate whether Huaier aqueous extract, a Chinese medicine, has efficacy against CSCs and to investigate the mechanisms of its anticancer effects. It was observed that the Huaier extract significantly inhibited the spheroid formation potential (P<0.05) and decreased the aldehyde dehydrogenase (ALDH)-positive cell population in colorectal primary cancer cells (P<0.05). Western blotting analysis and Wnt/β-catenin reporter assays revealed that the Huaier extract downregulated the Wnt/β-catenin self-renewal pathway. This is the first study to demonstrate that Huaier aqueous extract acts as an effective agent for eradicating colorectal CSCs and identifies the Wnt/β-catenin pathway as its potential target, which may be a new approach for colorectal cancer therapy.
Colonoscopy and fecal immunochemical testing (FIT) are accepted strategies for colorectal-cancer screening in the average-risk population.
In this randomized, controlled trial involving asymptomatic adults 50 to 69 years of age, we compared one-time colonoscopy in 26,703 subjects with FIT every 2 years in 26,599 subjects. The primary outcome was the rate of death from colorectal cancer at 10 years. This interim report describes rates of participation, diagnostic findings, and occurrence of major complications at completion of the baseline screening. Study outcomes were analyzed in both intention-to-screen and as-screened populations.
The rate of participation was higher in the FIT group than in the colonoscopy group (34.2% vs. 24.6%, P<0.001). Colorectal cancer was found in 30 subjects (0.1%) in the colonoscopy group and 33 subjects (0.1%) in the FIT group (odds ratio, 0.99; 95% confidence interval [CI], 0.61 to 1.64; P=0.99). Advanced adenomas were detected in 514 subjects (1.9%) in the colonoscopy group and 231 subjects (0.9%) in the FIT group (odds ratio, 2.30; 95% CI, 1.97 to 2.69; P<0.001), and nonadvanced adenomas were detected in 1109 subjects (4.2%) in the colonoscopy group and 119 subjects (0.4%) in the FIT group (odds ratio, 9.80; 95% CI, 8.10 to 11.85; P<0.001).
Subjects in the FIT group were more likely to participate in screening than were those in the colonoscopy group. On the baseline screening examination, the numbers of subjects in whom colorectal cancer was detected were similar in the two study groups, but more adenomas were identified in the colonoscopy group. (Funded by Instituto de Salud Carlos III and others; ClinicalTrials.gov number, NCT00906997.).
In the National Polyp Study (NPS), colorectal cancer was prevented by colonoscopic removal of adenomatous polyps. We evaluated the long-term effect of colonoscopic polypectomy in a study on mortality from colorectal cancer.
We included in this analysis all patients prospectively referred for initial colonoscopy (between 1980 and 1990) at NPS clinical centers who had polyps (adenomas and nonadenomas). The National Death Index was used to identify deaths and to determine the cause of death; follow-up time was as long as 23 years. Mortality from colorectal cancer among patients with adenomas removed was compared with the expected incidence-based mortality from colorectal cancer in the general population, as estimated from the Surveillance Epidemiology and End Results (SEER) Program, and with the observed mortality from colorectal cancer among patients with nonadenomatous polyps (internal control group).
Among 2602 patients who had adenomas removed during participation in the study, after a median of 15.8 years, 1246 patients had died from any cause and 12 had died from colorectal cancer. Given an estimated 25.4 expected deaths from colorectal cancer in the general population, the standardized incidence-based mortality ratio was 0.47 (95% confidence interval [CI], 0.26 to 0.80) with colonoscopic polypectomy, suggesting a 53% reduction in mortality. Mortality from colorectal cancer was similar among patients with adenomas and those with nonadenomatous polyps during the first 10 years after polypectomy (relative risk, 1.2; 95% CI, 0.1 to 10.6).
These findings support the hypothesis that colonoscopic removal of adenomatous polyps prevents death from colorectal cancer. (Funded by the National Cancer Institute and others.).
In a screen for gene copy-number changes in mouse mammary tumors, we identified a tumor with a small 350-kb amplicon from a region that is syntenic to a much larger locus amplified in human cancers at chromosome 11q22. The mouse amplicon contains only one known gene, Yap, encoding the mammalian ortholog of Drosophila Yorkie (Yki), a downstream effector of the Hippo(Hpo)-Salvador(Sav)-Warts(Wts) signaling cascade, recently identified in flies as a critical regulator of cellular proliferation and apoptosis. In nontransformed mammary epithelial cells, overexpression of human YAP induces epithelial-to-mesenchymal transition, suppression of apoptosis, growth factor-independent proliferation, and anchorage-independent growth in soft agar. Together, these observations point to a potential oncogenic role for YAP in 11q22-amplified human cancers, and they suggest that this highly conserved signaling pathway identified in Drosophila regulates both cellular proliferation and apoptosis in mammalian epithelial cells.
The Hippo pathway plays a key role in organ size control by regulating cell proliferation and apoptosis in Drosophila. Although recent genetic studies have shown that the Hippo pathway is regulated by the NF2 and Fat tumor suppressors, the physiological regulations of this pathway are unknown. Here we show that in mammalian cells, the transcription coactivator YAP (Yes-associated protein), is inhibited by cell density via the Hippo pathway. Phosphorylation by the Lats tumor suppressor kinase leads to cytoplasmic translocation and inactivation of the YAP oncoprotein. Furthermore, attenuation of this phosphorylation of YAP or Yorkie (Yki), the Drosophila homolog of YAP, potentiates their growth-promoting function in vivo. Moreover, YAP overexpression regulates gene expression in a manner opposite to cell density, and is able to overcome cell contact inhibition. Inhibition of YAP function restores contact inhibition in a human cancer cell line bearing deletion of Salvador (Sav), a Hippo pathway component. Interestingly, we observed that YAP protein is elevated and nuclear localized in some human liver and prostate cancers. Our observations demonstrate that YAP plays a key role in the Hippo pathway to control cell proliferation in response to cell contact.
TAZ is a WW domain containing a transcription coactivator that modulates mesenchymal differentiation and development of multiple
organs. In this study, we show that TAZ is phosphorylated by the Lats tumor suppressor kinase, a key component of the Hippo
pathway, whose alterations result in organ and tissue hypertrophy in Drosophila and contribute to tumorigenesis in humans. Lats phosphorylates TAZ on several serine residues in the conserved HXRXXS motif
and creates 14-3-3 binding sites, leading to cytoplasmic retention and functional inactivation of TAZ. Ectopic expression
of TAZ stimulates cell proliferation, reduces cell contact inhibition, and promotes epithelial-mesenchymal transition (EMT).
Elimination of the Lats phosphorylation sites results in a constitutively active TAZ, enhancing the activity of TAZ in promoting
cell proliferation and EMT. Our results elucidate a molecular mechanism for TAZ regulation and indicate a potential function
of TAZ as an important target of the Hippo pathway in regulating cell proliferation tumorigenesis.
Alzheimer's disease (AD) is the most common neurodegenerative disorder, characterized by progressive neuronal loss with amyloid β-peptide (Aβ) plaques. Despite several drugs currently used to treat AD, their beneficial effects on AD progress remains under debate. Here, we established a rapid in vivo screening system using Drosophila AD models to assess the neuroprotective activities of medicinal plants that have been used in traditional Chinese medicine. Among 23 medicinal plants tested, the extracts from five plants, Coriandrum sativum, Nardostachys jatamansi, Polygonum multiflorum (P. multiflorum), Rehmannia glutinosa, and Sorbus commixta (S. commixta), showed protective effects against the Aβ42 neurotoxicity. We further characterized the neuroprotective activity of ethanol extracts from P. multiflorum and S. commixta. Aβ42-expressing flies that we used showed AD neurological phenotypes, such as decreased survival and motility and increased cell death and reactive oxygen species level. However, feeding these flies extracts from P. multiflorum or S. commixta showed strong suppression of such phenotypes. Similar results were observed in human cells, so that the treatment of P. multiflorum and S. commixta extracts increased the viability of Aβ-treated SH-SY5Y cells. Moreover, 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside, one of the main constituents of P. multiflorum, also showed similar protective activity against Aβ42 cytotoxicity in both Drosophila and human cells. Taken together, our results suggest that both P. multiflorum and S. commixta have therapeutic potential for the treatment of neurodegenerative diseases, such as AD.
Hepatocellular carcinoma (HCC) is one of leading causes of death in the world. Although various treatments have been developed, the therapeutic side effects are far from desirable. Chinese medicines (CMs, including plants, animal parts and minerals) have drawn a great deal of attention in recent years for their potential in the treatment of HCC. Most studies have shown that CMs may be able to retard HCC progression with multiple actions, either alone or in combination with other conventional therapies to improve quality of life in HCC patients. Additionally, CMs are used for preventing HCC occurrence. The aim of this study is to review the potential prophylactic and curative effects of CMs on human HCC and the possible mechanisms that underlie these pharmacological actions. Publications were collected and reviewed from PubMed and China National Knowledge Infrastructure from 2000 to 2014. Keywords for literature searches include "Chinese medicine", "Chinese herb", "traditional Chinese Medicine", "hepatocellular carcinoma" and "liver cancer". CMs in forms of pure compounds, isolated fractions, and composite formulas are included. Combination therapies are also considered. Both in vitro and in vivo efficacies of CMs are being discussed and the translational potential to bedside is to be discussed with clinical cases, which show the actions of CMs on HCC may include tumor growth inhibition, antimetastatic activities, anti-inflammation, anti-liver cancer stem cells, reversal on multi-drug resistance and induction/reduction of oxidative stress. Multiple types of molecules are found to contribute in the above actions. The review paper indicated that CMs might have potential to both prevent HCC occurrence and retard HCC progression with several molecular targets involved.
Amyotrophic Lateral Sclerosis (ALS) is a progressive neuromuscular disease for which there is no cure. We have previously developed a Drosophila model of ALS based on TDP-43 that recapitulates several aspects of disease pathophysiology. Using this model, we designed a drug screening strategy based on the pupal lethality phenotype induced by TDP-43 when expressed in motor neurons. In screening 1200 FDA-approved compounds, we identified the PPARγ agonist pioglitazone as neuroprotective in Drosophila. Here, we show that pioglitazone can rescue TDP-43-dependent locomotor dysfunction in motor neurons and glia but not in muscles. Testing additional models of ALS, we find that pioglitazone is also neuroprotective when FUS, but not SOD1, is expressed in motor neurons. Interestingly, survival analyses of TDP or FUS models show no increase in lifespan, which is consistent with recent clinical trials. Using a pharmacogenetic approach, we show that the predicted Drosophila PPARγ homologs, E75 and E78, are in vivo targets of pioglitazone. Finally, using a global metabolomic approach, we identify a set of metabolites that pioglitazone can restore in the context of TDP-43 expression in motor neurons. Taken together, our data provide evidence that modulating PPARγ activity, although not effective at improving lifespan, provides a molecular target for mitigating locomotor dysfunction in TDP-43 and FUS but not SOD1 models of ALS in Drosophila. Furthermore, our data also identify several ‘biomarkers’ of the disease that may be useful in developing therapeutics and in future clinical trials.
Trametes robiniophila Murr. (Huaier) is a sandy beige mushroom found on the trunks of trees and has been widely used in traditional Chinese medicine (TCM) for ~1,600 years. The anticancer effects of Huaier have attracted increasing worldwide interest in recent years. Accumulating evidence suggests that the anticancer mechanism of Huaier may be associated with various biological activities, such as inhibition of cell proliferation, anti-metastasis, interference with tumor angiogenesis and tumor-specific immunomodulatory effect. Animal and experimental studies suggest that Huaier is a promising anticancer agent. Further clinical research is warranted to illustrate the untapped chemopreventive and therapeutic potential of Huaier either alone or in conjunction with existing therapies.
This study was carried out to evaluate the effects of a Huaier polysaccharide (TP-1) on the tumor growth and immune function in hepatocellular carcinoma (HCC) H22-based mouse in vivo. Results showed that TP-1 was capable of repressing transplanted H22 solid hepatic tumor cell growth in vivo, prolonging the live time of mice bearing ascetic H22 tumors, and repressing the pulmonary metastasis of H22-bearing mice. Moreover, the relative weight of immune organ (spleen and thymus) and lymphocyte proliferation were improved after TP-1 treatment. Furthermore, the treatment with TP-1 could promote immune-stimulating serum cytokines, such as IL-2 and IFN-γ, but inhibit immune-suppressing serum cytokines IL-10 secretion in H22-bearing mice. Besides, the percentage of CD4+ T cells and NK cells was increased, whereas the number of CD8+ T cells decreased in tumor-bearing mice following TP-1 administration. In addition, this compound displayed little toxic effects to major organ of tumor-bearing mice at the therapeutic dose, such as the liver and kidney. This experimental finding suggested that TP-1 exhibited prominent antitumor activities in vivo via enhancement of host immune system function in H22 tumor-bearing mice. This product could be developed individually as a safe and potent biological response modifier for HCC therapy.
The Hippo signaling pathway, consisting of a highly conserved kinase cascade (MST and Lats) and downstream transcription coactivators (YAP and TAZ), plays a key role in tissue homeostasis and organ size control by regulating tissue-specific stem cells. Moreover, this pathway plays a prominent role in tissue repair and regeneration. Dysregulation of the Hippo pathway is associated with cancer development. Recent studies have revealed a complex network of upstream inputs, including cell density, mechanical sensation, and G-protein-coupled receptor (GPCR) signaling, that modulate Hippo pathway activity. This review focuses on the role of the Hippo pathway in stem cell biology and its potential implications in tissue homeostasis and cancer.
We have recently reported that Astrocyte elevated gene-1 (AEG-1) might be an epithelial-mesenchymal transition (EMT) associated biomarker in human hepatocellular carcinoma (HCC), and play an important role in the progression of hepatocellular carcinoma. To extend our study, we examined here the anti-invasive and metastatic effects of Huaier polysaccharide (HP) on human HCC cell line MHCC97-H and explored its possible mechanism of action. Treatment with HP dose-dependently inhibited the proliferation, adhesion, migration and invasion of MHCC97-H cells in vitro. It achieved this by reducing the expression of AEG-1 and N-cadherin, but enhancing E-cadherin expression. Therefore, these data suggested that HP can inhibit the growth and metastatic potential of MHCC97-H cells through modulation of the AEG-1/EMT pathway.
The Hippo pathway controls organ size in diverse species, whereas pathway deregulation can induce tumours in model organisms and occurs in a broad range of human carcinomas, including lung, colorectal, ovarian and liver cancer. Despite this, somatic or germline mutations in Hippo pathway genes are uncommon, with only the upstream pathway gene neurofibromin 2 (NF2) recognized as a bona fide tumour suppressor gene. In this Review, we appraise the evidence for the Hippo pathway as a cancer signalling network, and discuss cancer-relevant biological functions, potential mechanisms by which Hippo pathway activity is altered in cancer and emerging therapeutic strategies.
A neutral water-soluble polysaccharide (W-NTRP), with a molecular weight of 2.5×10(4)Da, was isolated from the fruit bodies of Trametes robiniophila (Huaier). Gas chromatography (GC) results indicated that W-NTRP was determined to be galactose (Gal), arabinose (Ara) and glucose (Glc), with a relative molar ratio of 4.2:2.5:0.7. Its antitumor and immunomodulatory activity were evaluated in vitro. W-NTRP showed remarkable inhibitory effect on three human cholangiocarcinoma cell lines (QBC939, Sk-ChA-1 and MZ-ChA-1), with respective IC(50) values of 47.8, 75.9, and 43.7μg/mL, but had no cytotoxicity to L-929 normal cells. Furthermore, W-NTRP had proliferation promoting effect on mouse splenocytes with or without concanavalin A (ConA) or lipopolysaccharide (LPS) in a bell-shaped dose-response manner. In addition, W-NTRP could prominently stimulate macrophages to produce nitric oxide (NO) through the up-regulation of inducible NO synthase (iNOS) activity. These results suggest that W-NTRP could be explored as a potential antitumor agent for cholangiocarcinoma.
Expression changes in subsets of genes occur in the course of altering cell fates, i.e., aging, cell death, and carcinogenesis. These changes simultaneously provide the good candidate as a biomarker for monitoring cancer. We have identified a novel human septin family gene, Bradeion, from adult brain cDNA library by a monoclonal antibody CE5. Northern blot and in situ hybridization analysis showed that Bradeion has two distinct transcripts, approximately 2.2 and 1.7 kb length (α and β, respectively) mainly in brain and slightly in heart, and no expression in any fetal organs. Haplotype analysis placed the gene location at 17q23. The gene contains GTPase motifs highly conserved in the septin family genes that are essential for cytokinesis and cell separation. The transcript of β form lacks a hydrophobic region, which suggests that this form arises from a single Bradeion gene through unique RNA splicing. Interestingly, this brain-specific Bradeion gene is also expressed in two human cancers, colorectal cancer and malignant melanoma. Ectopic expression of normal Bradeion α and β transcripts were confirmed both in patients' tumor samples and in in vitro cultured human cancer cell lines. Thus the Bradeion provides valuable tools as a tumor-specific and selective marker.
Gas chromatography coupled to mass spectrometry (GC/MS) is a core analytical method for metabolomics and has been used as a platform in non-targeted analysis, especially for hydrophilic metabolites. Non-targeted GC/MS-based metabolomics generally requires a high-throughput technology to handle a large volume of samples and an accumulated database (reference library) of the retention times and mass spectra of standard compounds for accurate peak identification. In this study, we provide a practical GC/MS platform and an auto peak identification technique that is not restricted to certain types of mass spectrometers. The platform utilizes a quadrupole mass spectrometer capable of high-speed scanning, resulting in greater output compared with Pegasus GC-time of flight (TOF)/MS, which has been an essential instrument for high-throughput experiments. Moreover, we show that our reference library is broadly applicable to other instruments; peak identification can be readily performed using the library without constructing a reference resource. The usefulness and versatility of our system are demonstrated by the analyses of three experimental metabolomics data sets, including standard mixtures and real biological samples.
Aqueous extract of Trametes robiniophila murr (Huaier) has been commonly used in China for cancer complementary therapy in recent years; however, the mechanisms of its anticancer effects are largely unknown. In the present study, we aim to investigate its inhibitory effect on both MCF-7 and MDA-MB-231 cells, and explore the possible mechanisms of its anticancer effect. Cell viability and motility were measured by MTT and invasive assays, migration and scratch assays in vitro, respectively. The distribution of cell cycle, PI-Annexin-V staining and Rhodamine 123 assay were analyzed by flow cytometry, and western blot were used to test the apoptotic pathways. We found that Huaier extract could strongly inhibit cell viability of MCF-7 and MDA-MB-231 cells in a time- and dose-dependent manner; however, MDA-MB-231 cells showed more susceptibility to the treatment. Furthermore, cell invasiveness and migration were also suppressed with exposure to Huaier extract. We also indicated that Huaier could induce G0/G1 cell-cycle arrest, p53 accumulation and activation selectively in MCF-7 cells. Inspiringly, the PI-Annexin-V staining assay and western blot analysis confirmed cell apoptosis executed by caspase-3. Decreased mitochondrial membrane potential by Rhodamine 123 assay and down-regulation of Bcl-2 and up-regulation of BCL2-associated X protein (BAX) indicated that Huaier induced apoptosis through the mitochondrial pathway. Caspase activation during Huaier-induced apoptosis was confirmed by pan-caspase inhibitor, Z-VAD-fmk. As expected, the inhibitor decreased Huaier-induced apoptosis in both cell lines. Based on our findings, Huaier can induce cell apoptosis in both ER-positive and ER-negative breast cancer cell lines and is an effective complementary agent for breast cancer treatment.
The co-activator Yorkie (Yki) mediates transcriptional regulation effected by the Drosophila Fat-Warts (Wts)-Hippo (Hpo) pathways. Yki is inhibited by Wts-mediated phosphorylation, and a Wts phosphorylation site at Ser168 has been identified. Here we identify two additional Wts phosphorylation sites on Yki, and examine the respective contribution of all three sites to Yki nuclear localization and activity. Our results show that although Ser168 is the most critical site, all three phosphorylation sites influence Yki localization and activity in vivo, and can be sites of regulation by Wts. Thus, investigations of the role of Yki and its mammalian homolog Yes-associated protein (YAP) in development and oncogenesis should include evaluations of additional sites. The WW domains of Yki are not required for its phosphorylation, but instead are positively required for its activity. We also identify two potential sites of phosphorylation by an unknown kinase, which could influence phosphorylation of Ser168 by Wts, suggesting that there are additional mechanisms for regulating Yki/YAP activity.
Many pharmaceutical agents have been discovered by screening natural products from plants, animals, marine organisms and microorganisms. Vincristine, irinotecan, etoposide and paclitaxel are examples of plant-derived compounds that are being employed in cancer treatment, and dactinomycin, bleomycin and doxorubicin are anticancer agents derived from microbial sources. Citarabine is an example of an anticancer agent originating from a marine source. Other agents originating from marine sources are bryostatin-1, aplidine, dolastatin 10 and ET-743, which have recently entered phase I and II clinical trials.
Malignant tumor progression is a complex and multi-gene event which can not be easily detected or predicted. The detection of malignant cells using marker genes is hampered by the fact that these markers are only expressed by certain malignancies or lack sensitivity and/or specificity. We have reported a human septin family gene Bradeion, which shows strong cancer-specific expression in colorectal and urologic cancers as a result of carcinogenesis.
Diagnostic efficacy and validity of Bradeion gene expression were tested by two independent systems, one is a protein detection method using monoclonal antibody based immuno-chromatographic membrane strip tests (a nitrocellulose test strip assay), and another is a gene expression detection method, quantitative RT-PCR. The technology has been established using Bradeion fusion proteins, in vitro cultivated human cancer cell lines, and also patients' test samples with controls.
Bradeion test strip by combination with two monoclonal antibodies are valid for the detection of 1 ng/ml Bradeion, and successfully applied for patient urine samples with no false-positive results. Positive detection rates were over 70% of the patient urine samples so far tested (prostate cancer, renal cell carcinoma, and bladder cancer) in 15 to 30 minutes. Quantitative RT-PCR resulted in significantly high copy numbers of 0.4-3.0-3.0 x 10(5) per microg total RNA in patients' tissue samples, whereas those from normal tissue or other cancers found negative.
The present study introduces the practical diagnostic methods using a disease-specific molecular marker, which provides safe, economical, and rapid clinical screening of cancer.