组蛋白去乙酰化酶1对肝癌细胞增殖凋亡的影响

Effects of HDAC1 on proliferation and apoptosis of hepatocellular carcinoma

目的探讨组蛋白去乙酰化酶1(HDAC1)对肝癌细胞增殖凋亡的影响。方法肝癌细胞SNU-449转染HDAC1小干扰RNA(HDAC1 siRNA)和小干扰RNA阴性对照(siRNA control),同时设置不转染的细胞为对照。采用RT-PCR检测细胞中HDAC1mRNA的表达水平,Western blot检测细胞中HDAC1、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、信号转导与转录因子3(STAT3)、磷酸化的STAT3(p-STAT3)蛋白表达水平,噻唑蓝(MTT)检测细胞增殖情况,流式细胞术检测细胞凋亡情况。结果肝癌细胞转染siRNA control后,细胞存活率、凋亡率及细胞中HDAC1、Cleaved Caspase-3、STAT3、p-STAT3表达水平与对照细胞相比无显著差异(P>0.05)。肝癌细胞转染HDAC1 siRNA后,细胞中HDAC1mRNA和蛋白表达水平与对照细胞相比均受到抑制,并且细胞存活率只有(63.74±8.37)%,而细胞凋亡率升高为(32.84±6.92)%,细胞中Cleaved Caspase-3蛋白水平升高,p-STAT3蛋白水平降低。结论干扰HDAC1表达能够促进肝癌细胞凋亡,抑制肝癌细胞增殖,作用机制可能与STAT3信号通有关。

Objective To investigate the effects of histone deacetylase 1 （HDAC1） on proliferation and apoptosis of human hepatocellular carcinoma cells. Methods Hepatocellular carcinoma cell line SNU -4 49 was transfected with HDAC1 small interfering RNA （HDAC1 siRNA） and small interfering RNA negative control （siRNA control） , at the same time, the non transfected cells were used as controls, RT - PCR was used to detect the expression level of HDAClmRNA in cells, the expression levels of HDAC1 , Cleaved Caspase - 3 , STAT3 and p - STAT3 in the cells were detected by Western blot, cell proliferation was detected by MTT, cell apoptosis was detected by flow cytometry. Results After hepatocellu-lar carcinoma cells were transfected with siRNA control, the cell survival rate, apoptosis rate and the expression levels of HDAC1, Cleaved Caspase - 3 , STAT3 and p - STAT3 in the cells were not significantly different with those in the control group （ P 〉 0. 0 5 ） . After the hepatocellu-lar carcinoma cells were transfected with HDAC1 siRNA, the expression levels of HDAClmRNA and protein in cells were inhibited compared with control cells, and the cell survival rate was only （63 ？ 74 ± 8 ？ 37 ） %,the apoptosis rate was increased （32 . 84 ± 6 ？ 92 ） % ,levels of Cleaved Caspase -3 protein in cells elevated, p - STAT3 protein level decreased. Conclusion Interference of HDAC1 expression can promote apoptosis of hepatocellular carcinoma cells, inhibiting proliferation of hepatoma cells, the mechanism of action may be related to STAT3 signal transduction.