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Effect of Genistein in Comparison with trichostatin a on Reactivation of DNMTs Genes in Hepatocellular Carcinoma

Journal of Clinical and Translational Hepatology 2018年第2期6卷 141-146页

作者： Masumeh Sanaei Fraidoon Kavoosi Abazar Roustazadeh Fatemeh Golestan Research Center for Non-Communicable Diseases Jahrom University of Medical SciencesJahromFars provinceIran Student Research Committee Jahrom University of Medical SciencesJahromFars provinceIran

Background and Aims:DNA methylation and histone mod-ification are epigenetic modifications essential for normal function of mammalian *** processes are mediated by biochemical interactions between DNA methyltransferases(DNMTs)and histone *** hypermethyla-tion and deacetylation of tumor suppressor genes play major roles in cancer induction,through transcriptional silencing of these *** hypermethylation is carried out by a family of DNMTs including DNMT1,DNMT3a and *** hepatocellular carcinoma,a significant positive correlation bet-ween over-expression of these genes and cancer induction has been *** DNA demethylating agent genistein(GE)has been demonstrated to reduce different ***-viously,we reported that GE can induce apoptosis and inhibit proliferation in hepatocellular carcinoma PLC/PRF5 and HepG2 cell ***,histone deacetylase inhibitors,such as trichostatin a(TSA),were successfully used to inhibit cancer cell *** present study was designed to assess the effect of GE in comparison with TSA on DNMT1,DNMT3a and DNMT3b gene expression,cell growth inhibition and apoptosis induction in the HepG2 cell ***:Cells were seeded and treated with various doses of GE and *** MTT assay,flow cytometry assay,and real-time RT-PCR were used to determine viability,apoptosis,and DNMT1,DNMT3a and DNMT3b gene expression ***:Both agents inhibited cell growth,induced apoptosis and re-activated DNMT1,DNMT3a and DNMT3b gene ***,TSA demonstrated a significantly greater apop-totic effect than the other agent,whereas GE improved gene expression more significantly than ***:Our findings suggest that GE and TSA can significantly inhibit cell growth,induce apoptosis and restore DNMT1,DNMT3a and DNMT3b gene reactivation.

关键词： Genistein trichostatin a DNA methyltransferases Apoptosis Hepatocellular carcinoma

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Microarray Study of Mechanism of trichostatin a Inducing Apoptosis of Molt-4 Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2009年第4期29卷 445-450页

作者：洪振亚韩志强肖敏杨漾夏曦周剑峰 Department of Hematology Department of Obstetrics and Gynecology Tongji HospitalTongji Medical College Huazhong University of Science and Technology

Histone deacetylase was overexpressed in a variety of cancers and was closely correlated with oncogenic factors. The histone deacetylase inhibitor, trichostatin a （TSA） was shown to induce apoptosis in many cancer cells. However, the mechanism of TSA on induction of cancer cells apoptosis is poorly understood. This study was designed to characterize the global gene expression profiles before and after treatment of human leukemia cell line Molt-4 with TSA. Flow cytometry, MTT and DNA ladder were used to observe the effect of TSA on the apoptosis of MOLT-4 cells and normal human peripheral blood mononuclear cells （PBMC）. Microarray, reverse transcription-polymerase chain reaction （RTopCR） and Western blotting were used to detect the difference of gene and protein expressions of Molt-4 cells after incubation of the cells with TSA. The results showed that TSA could induce Molt-4 apoptosis in dose- and time-dependent manners but spared PBMCs. Microarray analysis showed that after incubation with TSA for 9 h, 310 genes were upregulated and 313 genes were deregulated. These genes regulate the growth, differentiation and survival of cells. Among these genes, STATSA was down-regulated by 80.4% and MYC was down-regulated by 77.3%. It was concluded that TSA has definite growth-inhibiting and apoptosis-inducing effects on Molt-4 cells in time- and dose-dependent manners, with weak cytotoxic effects on PBMCs at the same time. The mechanism of TSA selectively inducing apoptosis and inhibiting growth may be ascribed to the changes of pro-proliferation genes and anti-apoptosis genes.

关键词： trichostatin a apoptosis MOLT-4 cells microarray difference in gene expression

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Cell-cycle-dependent variation in UV absorption spectrum of Hela cells treated with trichostatin a

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The Chinese-German Journal of Clinical Oncology 2012年第12期11卷 737-740页

作者： Fengqiu Zhang Xiaoxia Wang Zhanguo Yang Physical Engineering College Zhengzhou University Zhengzhou 450052 China Henan Province Institute oF Metrology Zhengzhou450008.China

Objective： The aim of our study was to discovery the different cell cycle arrest effect after different densities HeLa cells treated with trichostatin a （TSA）. In addition, this study would find some important relationship between cycle arrest effect and UV absorption spectrum of cell. Methods： 0.2 IJM TSA was applied to act on HeLa cells of different density. Then, the cycle arrest effect and UV absorption spectrum of cells were investigated, which provide support to analyze the effect of TSA on cancer cells. Results： Cell cycle arrest effect in G0/G1 of the lower density cells was more obvious than that in other groups. The other discovery in this work was that the cellular UV absorption value was higher when the density of cultured cell was lower. Conclusion： This experiment would guide the clinical study on early or late stage cancer patients in the future. On the other hand, this work indicates when cells were arrested in G0/G1 phase, the cellular absorption value increased at the same time, so UV absorption spectrum could characterize the change of cell cycle.

关键词： trichostatin a HeLa cells cell cycle UV absorption spectrum

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Anticancer Activities of trichostatin a on Maligant Lymphoid Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第5期26卷 538-541页

作者：孙春艳刘新月陈燕刘芳 Institute of Hematology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

The anticancer activity of trichostain A （TSA） on human B cell non-Hodgkin＇s lymphoma and its mechanism were explored, The effect of TSA on the growth of Raji cells and normal peripheral blood mononuclear cells （NPBMNC） was studied by MTr assay, The effect of TSA on the apoptosis of Raji cells and NPBMNC was studied by flow cytometry and TDT-mediated dUTP nick end labeling （TUNEL）. The effect of TSA on the cell cycle of Raji cells was studied by propidium iodide method. The results showed that TSA potently inhibited proliferation of Raji cells at microgram concentrations and induced apoptosis of Raji cells in a time-and concentration-dependent manner. Treatment with TSA induced accumulation of cells in G0/G1 or G2/M and a concomitant decrease of cell population in S phase. However, NPBMNC was less sensitive to the cytotoxic effect of TSA than Raji cells. It was concluded that TSA may inhibit the proliferation of Raji cells by regulating the cell cycle and inducing the cell apoptosis. Moreover, TSA demonstrates low toxicity in NPBMNC but selectively induces apoptosis of Raji cells.

关键词： trichostatin a Raji cell Apoptosis Cell cycle Cytotoxicity

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Effect and Comparison of Sodium Butyrate and trichostatin a on the Proliferation/Differentiation of K562

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第3期23卷 249-253页

作者：李春蕊刘文励孟凡凯黄伟周剑锋孙汉英冯永东 Department of Hematology

In order to explore the molecular mechanisms of sodium butyrate and trichostatin a on K562 cell proliferation/differentiation, K562 cells were grown in the absence or presence of sodium butyrate or trichostatin a The percentage of viable cells was determined by trypan blue exclusion Differentiation was determined by nitro-blue tetrazolium (NBT) reduction and cell surface adhesion molecules analyzed by FACS Cell cycle distribution was studied after DNA staining by propidium iodide Cell cycle regulatory proteins were detected by Western blot and reverse transcription-polymerase chain reaction The results showed that sodiun butyrate blocked cells mainly at the G0/G1 phase of the cell cycle, whereas trichostatin a arrested the cells at G2 phase Sodium butyrate could down-regulate the mRNA expression of cyclin D1, but not affect its protein expression; down-regulate the protein expression of cyclin D3, but not affect its mRNA expression trichostatin a showed similar effects on cyclin D1 and D3 as sodium butyrate Both sodium butyrate and trichostatin a could stimulate p21 expression of K562 cells at mRNA and protein levels It may be concluded that sodium butyrate and trichostatin a could promote the proliferation/differentiation of the K562 cells, which might be contributed to the induced expression of cyclin D3 and p21 proteins

关键词： sodium butyrate cyclin D p21 trichostatin a

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trichostatin a Induces Apoptosis by Inhibiting Telomerase Activity and Expression of Telomerase Reverse Transcriptase in HL-60 Cells

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Journal of Chinese Pharmaceutical Sciences 2006年第2期15卷 115-120页

作者：周咏明郭伟周浩李慧玉刘黎琼姚军霞郑金娥郭天南黄士昂华中科技大学同济医学院附属协和医院干细胞研究和应用中心湖北武汉430022 华中科技大学同济医学院附属同济医院内分泌科湖北武汉430030 华中科技大学同济医学院附属协和医院血液科湖北武汉430022

Aim To investigate the effects of trichostatin a （TSA） on telomerase activity and the expression of human telomerase reverse transcriptase （hTERT） during apoptosis in vitro and the mechanisms in HL-60 cells. Methods The proliferative activity of HL-60 cells was assessed by MTT assay. Cell apoptosis was analyzed by flow cytometry. Telomerase activity was examined by TRAP-ELISA. The expression of telomerase subunits was analyzed by RT-PCR. Results A time- and dose-dependent inhibition was detected in HL-60 cells treated with TSA. After treatment with 600 nmol· L^-1 TSA for 48 h, the apoptosis rate in HL-60 cells was 42. 6% and telomerase activity decreased 1.95 ± 0.25, 1.73 ± 0. 12, and 1.52 ± 0. 09 for 12, 24, and 48 h, respectively. The expression of hTERTmRNA decreased. No significant changes were observed in the expression of hTRmRNA and hTPI mRNA. Condusion TSA inhibits telomerase activity and induces apoptosis in HL-60 cells. The underlying mechanism may be related to the down-regulation of hTERT transcription.

关键词： trichostatin a apoptosis telomerase human telomerase reverse transcriptase

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The enhancement of Arsenic Trioxideby trichostatin a in inducing differentiation of HL-60 cells

The enhancement of Arsenic Trioxideby Trichostatin A in indu...

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第十届全国免疫学学术大会

作者： Gaofeng Bi Xia Ren Guanhua Song Lianlian Li Xiaoyu Zhang Guosheng Jiang Institute of Basic Medicine Shandong Academy of Medical Sciences

Objective:In order to study if trichostatin a facilitate Arsenic Trioxide to induce differentiation of HL-60 cells and its underlying ***:the alteration in the cell proliferation was determined by MTT *** reduction assay and CD11b analysis by FACS were used to detect the differentiation of HL60 *** cycle arrest was measured by *** expressions of m RNA of P21,P27 and Cyclin D3,Cyclin E in HL60 cells after exposure to TSA and(or)Arsenic Trioxide were evaluated by semi-quantitative ***:It was found that the combinationof TSAand Arsenic Trioxide could inhibit proliferation and induce differentiation of HL60 cells obviously than Arsenic Trioxide *** up-regulationof the m RNA expression of P21,P27 but down-regulation of the expression of Cyclin D3,Cyclin E were related to the differentiation of ***:TSA has synergistic effect with Arsenic Trioxide in inducing differentiation of HL60 and it’s associated with the change of P21,P27 and Cyclin D3,Cyclin E expression.

关键词： trichostatin a Arsenic Trioxide HL-60 cell induce differentiation synergistic effect

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Regulation of Histone Acetylation and Apoptosis by trichostatin in HL-60 Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第6期24卷 572-574页

作者：李新刚陈维凯谷俊侠崔国惠陈燕 The Institute of Hematology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China

Summary: In order to examine the strong anticancer action and low toxicity of trichostatin a (TSA), the effect of TSA was examined on the growth inhibition, acetylation of histone H_3 and apoptosis in HL-60 cells by employing MTT, immunocytochemical techniques, and Annexin-V-FITC/PI assay. Our results showed that TSA could inhibit proliferation of HL-60 cells in a time-and dose-dependent manner, and the IC_~50 at the 36th h was 100 ng/ml. The apoptosis-inducing effect of TSA on HL-60 cells was also time-and dose-dependent. But it didn't demonstrate apparent apoptosis induction in NPBMNCs within specific dose and time range. Both of the acetylation of histone H_3 in HL-60 cells and NPBMNCs increased significantly (P0.05). It is concluded that TSA can inhibit growth and induce apoptosis of HL-60 cells in a time-and dose-dependent manner, and is able to selectively induce apoptosis in HL-60 cells but does not respond in NPBMNCs under the same conditions. The difference of TSA between HL-60 cells and NPBMNCs can't be explained by the regulation of histone acetylation.

关键词： trichostatin a deacetylase inhibitor histone acetylation apoptosis HL-60 cells

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Effect of histone deacetylase inhibitor on proliferation of biliary tract cancer cell lines

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World Journal of Gastroenterology 2008年第16期14卷 2578-2581页

作者： Li-Ning Xu Xin Wang Sheng-Quan Zou Institute of Hepatobiliary Surgery of PLA GeneralHospital Beijing 100853 China Department of General Surgery of Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei Province China

AIM: To explore the effect of histone deacetylase inhibitor, trichostatin a (TSA) on the growth of biliary tract cancer cell lines (gallbladder carcinoma cell line and cholangiocarcinoma cell line) in vivo and in vitro, and to investigate the perspective of histone deacetylase inhibitor in its clinical application. METHODS: The survival rates of gallbladder carcinoma cell line (Mz-ChA-l cell line) and cholangiocarcinoma cell lines (QBC939, KMBC and OZ cell lines) treated with various doses of TSA were detected by methylthiazoy tetrazolium (MTT) assay. A nude mouse model of transplanted gallbladder carcinoma (Mz-ChA-l cell line) was successfully established, and changes in the growth of transplanted tumor after treated with TSA were measured. RESULTS: TSA could inhibit the proliferation of gallbladder carcinoma cell line (Mz-ChA-l cell line) and cholangiocarcinoma cell lines (QBC939, KMBC and OZ cell lines) in a dose-dependent manner. After the nude mouse model of transplanted gallbladder carcinoma (Mz- ChA-l cell line) was successfully established, the growth of cancer was inhibited in the model after treated with TSA. CONCLUSION: TSA can inhibit the growth of cholangiocarcinoma and gallbladder carcinoma cell lines in vitro and in vivo.

关键词： Biliary tract cancer Gallbladder carcinoma Cholangiocarcinoma Proliferation trichostatin a

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Histone deacetylase inhibitors inducing human cervical cancer cell apoptosis by decreasin~ DNA-methyltransferase 3B

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Chinese Medical Journal 2012年第18期125卷 3273-3278页

作者： LIU Ning ZHAO Li-jun LI Xiao-ping WANG Jian-liu CHAI Guo-lin WEI Li-hui Department of Obstetrics and Gynecology Peking UniversityPeople's Hospital China Beijing 100044 Department of Hematology China-Japan Friendship Hospital China Beijing 100029

Background Histone deacetylase （HDAC） inhibitors are a group of small chemical molecules that inhibit histone deacetylase. At cell level, HDAC inhibitors have multiple biological effects such as cell cycle arrest, apoptosis, cell differentiation and auotophagy. At molecular level, HDAC inhibitors cause histone and nonhistone acetylation and induce gene expression. HDAC inhibitors are widely used in cancer therapy because of its function of inducing apoptosis. However, the mechanisms of apoptosis effect are not fully understood. TSA is a classical HDAC inhibitor and widely used in epigenetic and anti-cancer research. In this study, we selected trichostatin a （TSA） to investigate the mechanisms of HDAC inhibitors apoptotic effect on cancer cells. Methods Cervical cancer cell lines such as Hela, Caski and normal human keratinocyte line HaCaT were treated with various concentrations of TSA. Crystal violent assay and 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay were performed to determine cell number. PARP cleavage and FITC-AnexinV were performed to determine apoptosis. DNA-methyltransferase （DNMT）I, DNMT3A and DNMT3B were determined by regular PCR, qPCR and Western Blotting. Small interfering RNA （SiRNAi） was used to knock down DNMT3B. Results HDAC inhibitors only induce cervical cancer cell apoptosis. At 1 Iumol/L of TSA, 86% of Hela cell and 76% of Caski went apoptosis. For normal cells, HDAC inhibitors have no cytotoxic effect at therapeutic dosage, （90.0＋8.4）% of normal cell survive after treated with 1 IJmol/L of TSA. We compared 1 pmol/L group with untreated control with t-test. There was no significance between 1 pmol/L group and untreated control for normal cell （P 〉0.05）. HDAC inhibitors decreased DNMT3B in cancer cell but not in normal cell. Manually knock-down of DNMT3B induced Hela and Caski cell apoptosis. More than 99% of Hela and Caski cell went apoptosis after deprived of DNMT3B. Conclusions DNMT3B was essential to cervical ca

关键词： histone deaeetylase inhibitors human cervical cancer apoptosis DNA-methyltransferase 3B trichostatin a DNA methyl transferase

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