TSA对人肝癌HepG2细胞增殖、凋亡及FHIT表达的影响

目的观察曲古抑菌素A（TSA）的体外抗人肝癌HepG2细胞作用及机制。方法取人肝癌HepG2细胞培养至对数生长期后随机分为TSA组、对照组、空白组,TSA组加入TSA至终浓度分别为125、250、500、1 000、2 000nmol/L,对照组加入等量二甲基亚砜（DMSO）,空白组只加培养基、无细胞,每组设6个复孔,作用24~72 h后MTT比色法测定HepG2细胞增殖抑制率（IR）,倒置显微镜和电镜观察HepG2细胞形态变化,TUNEL法检测细胞凋亡率,实时荧光定量PCR检测脆性组氨酸三联体（FHIT）基因表达,Western blot检测FHIT蛋白表达。结果与对照组相比,TSA组IR随TSA浓度增加和作用时间延长而增加（P〈0.05）,透射电镜下TSA组HepG2细胞出现凋亡早期改变;与对照组比较,TSA组细胞凋亡率升高,FHIT mRNA及蛋白表达增强（P均〈0.01）。结论 TSA体外能有效抑制肝癌细胞株HepG2生长并促进其凋亡,机制可能为上调FHIT表达。

Effect of TSA on the proliferation ,apoptosis and expression of FHIT in human hepatoma cell line HepG2

Objective To observe the effects of histone deacetylase inhibitor trichostatin A（TSA） on the growth of human hepatoma cell line HepG2 and its mechanism.Methods Human hepatoma HepG2 cells cultured in logarithmic phase were divided randomly into TSA group,control group and blank group,the TSA group was treated with TSA at the concentrations of 125,250,500,1 000,2 000 nmol/L,the control group was treated with equal volume of DMSO,and the blank group was treated with only medium without cells,24 to 72 hours later,the cell proliferation inhibition rate（IR） was analyzed by MTT assay,the cell morphology was observed by the inverted light microscope and electron-microscopy,and the apoptosis index was determined by using TUNEL technique.The expression of FHIT gene and protein was quantified using real-time PCR and Western blot.Results Compared with the control group,IR of the TSA group increased in dose and time-depended manner（P0.05）,early apoptosis of the HepG2 cells was observed under electron microscope;compared with the control group,the apoptosis rate of the TSA group increased,the expression of FHIT mRNA and protein increased（P0.01）.Conclusion TSA can inhibit the proliferation and promote apoptosis of hepatocarcinoma cell HepG2,the mechanism may relate to the up-regulating of FHIT expression.