曲古霉素A对肺腺癌细胞株NCI-H1299增殖的抑制作用及其机制

背景与目的：曲古霉素A（trichostatin A,TSA）是具有组蛋白去乙酰化酶（histone deacetylases,HDAC@强效非竞争性抑制剂,对血液系统肿瘤和实质性肿瘤均有较强的生长抑制作用。本文观察HDACs抑制剂TSA对体外培养的肺腺癌NCI-H1299细胞株的增殖、凋亡和周期以及相关基因表达的影响,并探讨其可能的作用机制。方法：MTT法检测不同浓度（0.1、0.2、0.4、2.0μmol/L@的TSA对人肺腺癌NCI-H1299细胞株体外增殖的影响,流式细胞术检测药物处理后细胞周期及凋亡率的变化;Western blot法检测细胞内组蛋白H4乙酰化水平的变化;Real-time PCR检测NCI-H1299细胞内p21、CyclinB1、Bcl-2和Bax的基因表达。结果：TSA能明显抑制NCI-H1299细胞的体外生长,其抑制作用呈明显的剂量和时间依赖性。TSA诱导后,流式细胞术检测结果显示细胞阻滞于G2/M期,细胞凋亡增加。TSA可明显提高NCI-H1299细胞内组蛋白H4的乙酰化水平,诱导p21和Bax的mRNA表达增加,同时抑制Bcl-2和CyclinB1表达。结论：TSA可通过诱导细胞凋亡及阻滞细胞周期而发挥体外抗肺腺癌细胞生长的作用,其机制可能与组蛋白乙酰化水平的提高以及调控相关基因p21、Bax、Bcl-2和CyclinB1的表达变化有关。     

Cell growth inhibition and gene expression induced by the histone deacetylase inhibitor, trichostatin A, on human hepatoma cells

OBJECTIVE: Histone deacetylase (HDAC) inhibitors have been reported to induce cell growth arrest, apoptosis and differentiation in tumor cells. The effect of the HDAC inhibitor, trichostatin A (TSA), on hepatoma cells, however, has not been well studied. In this study, we examined cell viability and gene expression profile in hepatoma cell lines treated with TSA. METHODS: To study cell growth inhibition and induction of apoptosis by TSA on human hepatoma cell lines including HuH7, Hep3B, HepG2, and PLC/PRF/5, cells were treated with TSA at various concentrations and analyzed by the 3-(4, 5-dimethyl-2-thiazolyl)-2H-tetrazolium bromide (MTT) and TUNEL assays, respectively. Changes in gene expression profile after exposure to TSA were assessed using a cDNA microarray consisting of 557 distinct cDNA of cancer-related genes. The levels of acetylated histones were examined by the chromatin immunoprecipitation (ChIP) assay using anti-acetylated histone H3 or H4 antibody. RESULTS: The MTT assay demonstrated that TSA showed cell growth inhibition not only in a concentration-dependent but also a time-dependent manner on all cell lines studied. The TUNEL assay also revealed the potential of TSA to induce apoptosis. The microarray analysis revealed that 8 genes including collagen type 1, alpha2 (COL1A2), insulin-like growth factor binding protein 2 (IGFBP2), integrin, alpha7 (ITGA7), basigin (BSG), quiescin Q6 (QSCN6), superoxide dismutase 3, extracellular (SOD3), nerve growth factor receptor (NGFR), and p53-induced protein (PIG11) exhibited substantial induction (ratio >2.0) after TSA treatment in multiple cell lines. ChIP assay, in general, showed a good correlation between the expression level of mRNA and levels of acetylated histones in these upregulated genes. CONCLUSIONS: This study showed cell growth inhibition and the gene expression profile in hepatoma cell lines exposed to TSA. The alteration in levels of acetylated histones was closely associated with expression of specific cancer-related genes in hepatoma cells.     

TRICHOSTATIN A INHIBITS PROLIFERATION, INDUCES APOPTOSIS AND CELL CYCLE ARREST IN HELA CELLS

Objective： The histone deacetylase inhibitors （HDACIS） have been shown to inhibit cancer cell proliferation, stimulate apoptosis, an induce cell cycle arrest. Our purpose was to investigate the antiproliferative effects of a HDACI, trichostatin A （TSA）, against human cervical cancer cells （HeLa）. Methods： HeLa cells were treated in vitro with various concentrations of TSA. The inhibitory effect of TSA on the growth of HeLa cells was measured by MTT assay. To detect the characteristic of apoptosis chromatin condensation, HeLa cells were stained with Hoechst 33258 in the presence of TSA. Induction of cell cycle arrest was studied by flow cytometry. Changes in gene expression of p53, p21wafl and p27Kipl were studied by semiquantitative RT-PCR. Results： TSA inhibited cell growth in a time- and dose-dependent manner. Hoechst 33258 staining assay showed that TSA induced apoptosis. Cell cycle analysis indicated that treatment with TSA decreased the proportion of cells in S phase and increased the proportion of cells in G0/G1 and/or G2/M phases of the cell cycle. This was concomitant with overexpression of genes related to malignant phenotype, including an increase in p53, p21wall and p27Kipl. Conclusion： These results suggest that TSA is effective in inhibiting growth of HeLa cells in vitro. The findings raise the possibility that TSA may prove particularly effective in treatment of cervical cancers.     

Histone Deacetylase Inhibitor Trichostatin A Enhances Antitumor Effects of Docetaxel or Erlotinib in A549 Cell Line

Background and Objective: Histone deacetylase (HDAC) inhibitors represent a promising class of potentialanticancer agents for treatment of human malignancies. In this study, we investigated the effect of trichostatinA (TSA), one such HDAC inhibitor, in combination with docetaxel (TXT), a cytotoxic chemotherapy agent orerlotinib, a novel molecular target therapy drug, on lung cancer A549 cells. Methods: A549 cells were treated withTXT, erlotinib alone or in combination with TSA, respectively. Cell viability, apoptosis, and cell cycle distributionwere evaluated using MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assay, Hochst33258staining and flow cytometry. Moreover, immunofluorescent staining and Western blot analysis were employedto examine alterations of α-tubulin, heat shock protein 90 (hsp90), epidermal growth factor receptor (EGFR),and caspase-3 in response to the different exogenous stimuli. Results: Compared with single-agent treatment,co-treatment of A549 cells with TSA/TXT or TSA/erlotinib synergistically inhibited cell proliferation, inducedapoptosis, and caused cell cycle delay at the G2/M transition. Treatment with TSA/TXT or TSA/erlotinib ledto a significant increase of cleaved caspase-3 expression, also resulting in elevated acetylation of α-tubulin orhsp90 and decreased expression of EGFR, which was negatively associated with the level of acetylated hsp90.Conclusions: Synergistic anti-tumor effects are observed between TXT or erlotinib and TSA on lung cancercells. Such combinations may provide a more effective strategy for treating human lung cancer.     

吉西他滨抑制骨肉瘤细胞系MG-63增殖和诱导凋亡的实验研究

目的研究吉西他滨对骨肉瘤细胞系MG-63的增殖抑制和凋亡诱导作用。方法利用光镜检测吉西他滨作用后MG-63的形态学变化;MTT法检测吉西他滨对MG-63的增殖抑制;流式细胞仪AnnexinV-FITC法检测细胞凋亡,琼脂糖凝胶电泳检测DNA ladder。结果吉西他滨可明显地抑制MG-63细胞生长,各实验组24、48、72小时抑制率随时间的增加而增加,差异有统计学意义（P〈0.05）,0.1 mg/L及以上浓度的吉西他滨对MG-63骨肉瘤细胞的抑制率可达50%,0.1-100 mg/L的吉西他滨对MG-63的抑制率相似。吉西他滨可诱导MG-63细胞凋亡,在不同浓度吉西他滨（0.1,1.0,10.0,100 mg/L）作用48小时后,MG-63的凋亡率差异无统计学意义（P〉0.05）。并且经琼脂糖凝胶电泳可以显示DNA ladder。结论吉西他滨对骨肉瘤细胞系MG-63具有增殖抑制作用,并且各实验组24、48、72小时抑制率随时间的增加而增加;吉西他滨对骨肉瘤细胞系MG-63具有诱导凋亡作用;0.1-100 mg/L吉西他滨对MG-63的增殖抑制和诱导凋亡作用相似。     

曲古抑霉素Ａ通过HIF-2α途径抗肿瘤作用的体内外实验研究

目的 探讨组蛋白去乙酰化酶（histone deacetylase,HDAC）抑制剂曲古抑霉素Ａ （trichostatin A,TSA）通过HIF-2α途径抗肿瘤作用机制。方法 体外模拟低氧环境培养骨肉瘤MG-63细胞,RT-PCR、western blot检测不同浓度的TSA对骨肉瘤MG-63细胞HIF-2α、VEGF及Glut-1 mRNA、蛋白表达的影响。使用蛋白酶体抑制剂MG132处理MG-63细胞和VHL基因缺陷的786-0细胞,western blot检测TSA对HIF-2α表达的影响。骨肉瘤MG-63细胞注入裸鼠皮下,待肿瘤体积长至20 mm3时,分为TSA组（N＝6）和生理盐水组（N＝6）,每３天给药１次,17天后镜下观察肿瘤组织形态;裸鼠处死后肿瘤组织提取蛋白,western blot检测HIF-2α表达情况。结果 体外低氧状态下,在培养的MG-63细胞系中,TSA呈浓度依赖性抑制HIF-2α蛋白及靶基因VEGF和Glut-1mRNA的表达;在蛋白酶体抑制剂MG132存在的情况下,可以逆转TSA对HIF-2α的抑制作用。在VHL基因缺陷的786-0细胞系中,TSA呈浓度依赖性抑制HIF-2α蛋白表达,MG132可以逆转TSA的作用。在动物移植瘤中,TSA抑制HIF-2α的表达。结论 实验表明,TSA通过一个VHL非依赖性的蛋白酶体依赖的途径抑制HIF-2α表达,TSA的抗肿瘤作用有一部分是通过抑制HIF-2α介导的,这为研究组蛋白去乙酰化酶抑制剂抗肿瘤作用的分子机制提供了一个新的方向。     

黄芩苷对骨肉瘤MG-63细胞增殖、凋亡的影响及其机制研究

目的:检测黄芩苷对骨肉瘤MG-63细胞增殖活性、凋亡指数、超微结构以及Survivin、VEGF、CAS-3蛋白表达水平的影响,评估黄芩苷对骨肉瘤的治疗学效应,并探讨其分子生物学机制。方法:骨肉瘤MG-63细胞培养成功后,分别采用0、50、100、200μg/ml浓度黄芩苷作用于骨肉瘤MG-63细胞。MTT法检测细胞增殖抑制率,流式细胞术检测细胞凋亡指数,透射电镜检测细胞超微结构改变,Western blot法检测Survivin、VEGF及CAS-3蛋白的表达。结果:MTT结果显示,黄芩苷对MG-63细胞的增殖有明显的抑制作用,且呈剂量和时间依赖性。流式细胞仪检测结果显示,治疗组中MG-63细胞凋亡指数显著高于对照组。透射电镜观察细胞超微结构,可见黄芩苷对MG-63超微结构有明显的影响。Western blot分析显示,随着黄芩苷浓度的增加,Survivin及VEGF蛋白的表达逐渐下降,而CAS-3表达增加,呈剂量依赖性。结论:黄芩苷可抑制骨肉瘤细胞增殖,诱导细胞凋亡,其机制可能是通过抑制肿瘤新生血管形成或通过调控CAS-3的表达、下调Survivin信号通路来完成的。     

洛铂对骨肉瘤MG-63细胞增殖、凋亡的影响及其机制

目的 探讨洛铂对骨肉瘤MG-63细胞增殖与凋亡的作用及其机制。方法 取对数生长期的MG-63细胞进行实验,分为对照组和实验组 (加入不同浓度洛铂: 2、4和8 μg/ml)。采用噻唑盐（MTT）比色法、流式细胞技术(FCM)检测洛铂对MG-63细胞的形态改变、生长及增殖抑制、细胞周期阻滞和细胞凋亡诱导等作用,并绘制不同浓度时MG-63细胞的生长曲线,应用Western blot的方法分析洛铂对MG-63细胞的Bcl-2蛋白表达的影响。结果 洛铂能够抑制MG-63细胞的生长、增殖并诱导其凋亡,并呈浓度和时间依赖效应。Western blot检测结果显示洛铂可使MG-63细胞Bcl-2蛋白表达下降。结论 洛铂能显著抑制MG-63细胞增殖,诱导细胞凋亡和细胞周期变化,可能与调节 MG-63细胞Bcl-2蛋白的表达有关。     

Induction of apoptosis by Trichostatin A in human breast cancer cell lines: involvement of 15-Lox-1

15-Lipoxygenase-1 (15-Lox-1) is a key enzyme mediating oxidative metabolism of polyunsaturated fatty acids and has attracted considerable interest as a potential target for the induction of apoptosis in cancer cells. Knowledge of relationship between 15-Lox-1 and histone deacetylase inhibitors is lacking in the breast cancer. This study is aimed to investigate the role of Trichostatin A (TSA) and 13(S)-HODE, as a metabolite of 15-Lox-1, in the regulation of breast cancer cell growth. The cytotoxic effect of TSA, as a potent HDAC inhibitor, was measured using MTT assay. Annexin V–FITC and PI staining were performed to detect apoptosis and cell cycle distribution using Flow cytometry. The role of 15-Lox-1 in the regulation of cell growth was assessed by 15-Lox-1 inhibitor and the level of 15-Lox-1 metabolite was measured to determine 15-Lox activity after treatment by TSA. The results demonstrated that TSA induced cell growth inhibition via 15-Lox-1, in a dose- and time-dependent manner, and subsequently accompanied by the cell cycle arrest and induction of apoptosis. Moreover, growth inhibitory effect of TSA was associated with the elevation of 15-Lox-1 metabolite (13(S)-HODE). This study provided evidences that the inhibitory effect of TSA on the breast cancer cell growth occurs via the induction of 15-Lox-1 activity and 13(S)-HODE production. Our findings underline the possible role of 15-Lox-1/13(S)-HODE pathway as a promising molecular approach for the induction of apoptosis in breast cancer cells.     

重组腺病毒载体CRT联合紫杉醇对乳腺癌细胞周期、增殖及凋亡的影响

目的:研究Ad-CRT联合紫杉醇对乳腺癌MDA-MB-231细胞周期、细胞增殖及凋亡的影响.方法:MTT法检测细胞增殖,相差显微镜观察细胞形态学变化;PI单染流式细胞术检测细胞周期和凋亡.结果:Ad-CRT联合紫杉醇对乳腺癌细胞MDA-MB-231增殖的抑制率明显高于单纯Ad-CRT组和单纯紫杉醇组,且抑制作用呈时间依赖关系(P<0.05),且细胞形态发生明显的改变.Ad-CRT联合紫杉醇将乳腺癌细胞MDA-MB-231的细胞周期明显的阻滞在G2/M期,并且促进细胞的凋亡.结论:Ad-CRT联合紫杉醇能增强乳腺癌细胞MDA-MB-231增殖抑制、阻滞细胞周期并且诱导细胞凋亡.     

低分子量肝素抑制骨肉瘤细胞Livin表达并诱导细胞凋亡的实验研究

Objective： To investigate the inhibitory effects of LMWH suppressing the expression of Livin and inducing the apoptosis of the osteosarcoma cells. Methods： Osteosarcoma cells line MG-63 was cultured in vitro. MTT assay and flow cytometry were used to study the effect of LMWH with different concentration suppressed the prolifetation and induced apoptosis in osteosarcoma cells line MG-63. The expression of Livin of osteosarcoma cells line MG-63 was analysed by the immunohistochemistrical method and PT-PCR. Results： Low molecular weight heparin could inhibit the growth of osteosarcoma cell line MG-63. With the LMWH＇s increasing, the apoptosis rate was increased significantly. Immunohistochemistrical method and PT-PCR showed that the expression of Livin of osteosarcoma cells line MG-63 declined obviously than that before medication. Conclusion： LMWH has very strong anti-tumor effect in vitro. The possible mechanisms of LMWH anti-tumor effect are associate with the effect of suppressing the expression of Livin and inducing cell apoptosis.     

番茄红素对人骨肉瘤细胞生长的影响及机制研究

目的 研究番茄红素(Lycopene,LP)对人骨肉瘤MG-63细胞增殖和凋亡的影响并初步探讨其机制。方法 取处于对数生长期的人骨肉瘤MG-63细胞,设空白对照组,LP低、中、高剂量组(5、10、20μg/mL)和顺铂(40μg/mL)组;给药干预48h,观察细胞生长状况,MTT比色法检测细胞增殖抑制率,流式细胞术(Flow cytometry,FCM)检测细胞周期、细胞凋亡状况并计算凋亡率,逆转录PCR(RT-PCR)法检测凋亡相关基因Bax mRNA、bcl-2 mRNA表达并计算Bax/bcl-2值,免疫蛋白印迹法(Western blot)检测凋亡相关蛋白caspase-3表达。结果 光学显微镜下观察可见,空白对照组人骨肉瘤MG-63细胞呈贴壁生长,增殖迅速,而LP组细胞增殖受到抑制,细胞数明显减少,呈现回缩、脱落现象。与空白对照组比较发现经LP干预48h能够显著提高人骨肉瘤MG-63细胞增殖抑制率,显著延长细胞周期G₀/G₁期并缩短S期、G₂/M期,显著提高细胞凋亡率(Apoptosis index,AI),上调Bax mRNA表达并下调bcl-2 mRNA表达,显著提高Bax/bcl-2比值,显著上调caspase-3蛋白表达,差异均具有统计学意义(P＜0.05或P＜0.01)。结论 LP具有抑制人骨肉瘤细胞增殖并促进其凋亡的药理学作用,机制可能与阻滞细胞周期和影响凋亡调控基因、蛋白表达有关。     

华蟾素和顺铂对骨肉瘤MG-63细胞联合杀伤作用的研究

目的：探讨比较华蟾素（ cinobufagin ）协同顺铂（ cisplatin ）对人骨肉瘤细胞株 MG-63的杀伤作用。方法体外培养骨肉瘤细胞株 MG-63细胞,取对数生长期细胞进行实验;采用 MTT 比色法测定MG-63细胞的生长抑制作用并计算细胞抑制率;流式细胞仪检测骨肉瘤细胞的凋亡率和细胞周期,观察细胞凋亡率的变化情况和药物对细胞周期的抑制作用;原位末端标记法（ TUNEL ）观察骨肉瘤细胞的凋亡情况并检测细胞凋亡率;相差显微镜观察骨肉瘤细胞生长状况及形态学上的改变。结果华蟾素和顺铂均对骨肉瘤细胞有生长抑制作用,华蟾素组、顺铂组及华蟾素＋顺铂组在药物浓度不同时对骨肉瘤细胞的生长抑制作用各不相同,在药物浓度低于80μg/ml时,各实验组药物对细胞的生长抑制作用随药物浓度的增加而上升;当药物浓度达到80μg/ml时,华蟾素组和顺铂组药物对骨肉瘤细胞的生长抑制作用不再增加,但华蟾素＋顺铂组药物对细胞的生长抑制作用在药物浓度达到160μg/ml时,抑制作用仍强于其浓度为80μg/ml时。各实验组在药物浓度相同时华蟾素＋顺铂对骨肉瘤细胞的生长抑制作用明显强于单独应用华蟾素或者顺铂。结论华蟾素＋顺铂对骨肉瘤 MG-63细胞的联合杀伤作用明显强于单独应用顺铂或者华蟾素;在抑制骨肉瘤 MG-63细胞生长方面,华蟾素协同顺铂对细胞的杀伤作用效果显著,并且具有时间和浓度依赖性。     

氧化砷对NB4和Jurkat细胞周期的影响

目的：研究氧化砷(As2O3)对白血病细胞系NB4、Jurkat细胞周期的影响及对细胞周期及凋亡相关蛋白的调节作用。方法：以MTT、基因组DNA电泳、蛋白／DNA双参数流式细胞术等方法研究As2O3对白血病细胞的作用及机制。结果：随着As2O3作用时间的延长，白血病细胞增殖明显受到抑制，DNA电泳出现“梯”状条带，流式细胞术检出亚G1峰，细胞周期阻滞于G1和G2／M期，部分细胞周期及凋亡相关的调控蛋白表达改变。结论：As2O3通过改变细胞周期及凋亡相关蛋白的表达而抑制白血病细胞增殖和诱导其凋亡。     

曲古抑菌素A通过p53转录激活途径诱导尤文肉瘤细胞凋亡

目的：探讨组蛋白去乙酰化酶抑制剂曲古抑菌素A（trichostatin A,TSA）诱导尤文肉瘤细胞株WE-68和VH-64凋亡及作用机制.方法：四甲基偶氮唑蓝法（MTT法）测定细胞增殖抑制率.流式细胞计数法测量TSA给药后细胞周期中sub-G1含量的变化.免疫印迹法（Western-blot）检测细胞中活化型多聚ADP核糖多聚酶（cleaved-PARP）,p53-lys382残基乙酰化和p53蛋白总量的表达.实时定量PCR和siRNA转染技术测定p53多个下游基因的mRNA水平改变和p53表达下调对TSA诱导凋亡的影响.结果：TSA抑制了尤文肉瘤细胞的增殖,诱导细胞周期中sub-G1含量和凋亡终产物cleaved-PARP蛋白表达的增加.TSA给药后,p53-lys382残基乙酰化表达量呈浓度依存性增加,同时上调p53下游因子p21,mdm2,Bax和PUMA的mRNA水平.另一方面,p53蛋白表达的下调明显削弱了TSA介导的p21表达的上调和cleaved-PARP的产生.结论：组蛋白去乙酰化酶抑制剂TSA能够通过激活p53高乙酰化表达来恢复p53转录功能,从而诱导尤文肉瘤细胞株产生凋亡.     

Cox-2抑制剂对骨肉瘤细胞抑制作用的实验研究

目的研究环氧化酶-2抑制剂Celecoxib对人骨肉瘤MG-63细胞的抑制作用及作用机制。方法用四唑盐（MTT）比色法检测不同浓度和不同作用时间对骨肉瘤细胞增殖的影响,用流式细胞仪检测药物对细胞周期和凋亡率的影响,透射电镜观察细胞的形态学改变。结果Celecoxlb能明显抑制细胞增殖,并呈剂量和时间依赖性。细胞凋亡率随药物浓度的增加逐渐增高,并引起细胞周期比例的改变。透射电镜显示细胞呈凋亡形态。结论Celecoxib对人骨肉瘤MG-63细胞有细胞毒作用,其机制与抑制肿瘤细胞增殖,使细胞阻滞于G0／G1期,并能诱导细胞凋亡有关。     

Histone deacetylase inhibitors such as sodium butyrate and trichostatin A induce apoptosis through an increase of the bcl-2-related protein Bad

The effects of sodium butyrate (SB) and trichostatin A (TSA) on cell proliferation and apoptosis against human glioma T98G, U251MG, and U87MG cells were investigated. Upon exposure to either SB or TSA, cell proliferation was reduced, and apoptosis detected by DNA fragmentation analysis and the cleavage of CPP32 was induced. Previously, we reported that SB increased the expression levels of p21 (WAF-1) and inhibited G1-S transition of the cell cycle. In this study, we showed that TSA also increased p21 expression, suggesting that histone deacetylase (HDAC) inhibitors may up-regulate p21 protein in common and thus arrest proliferation in the G1 phase of the cell cycle. To further determine the underlying molecular mechanisms of apoptosis with either SB or TSA treatment, we studied the expression levels of apoptosisrelated proteins in human glioma cells. SB increased the expression of the Bad protein, although the expression of Bcl-2, Bcl-xL, Bax, and Fas was not changed by the addition of SB. TSA treatment also up-regulated the expression of Bad protein. The results suggest that HDAC inhibitors such as SB and TSA induce apoptosis through an increase in Bad protein in human glioma cells in vitro.     

Effect of DNA Methyltransferase in Comparison to and in Combination with Histone Deacetylase Inhibitors on Hepatocellular Carcinoma HepG2 Cell Line

Background: DNA demethylating agents and histone deacetylase inhibitors can affect reactivation of geneexpression and apoptosis induction by DNA acetylation and demethylation. The aim of the present study was to analyzethe effects of DNA demethylating agent genistein (GE) and histone deacetylase inhibitor valproic acid VPA), aloneand combined, on hepatocellular carcinoma Hep G2 cell line. Methods: The cells were treated with various doses ofgenistein and valproic acid (alone and combined) and the MTT assay and flow cytometry were used to determine cellviability and apoptosis. Results: Genistein and valproic acid inhibited the growth of HepG 2 cells significantly. Resultof flow cytometry demonstrated that genistein and valproic acid (alone and combined) induce apoptosis significantly ina time‑dependent manner. Conclusions: Genistein and valproic acid can significantly inhibit proliferation and induceapoptosis in HepG2 cell line. The apoptotic effects of GE in combination with VPA were more significant that of eachcompound alone.