曲古菌素A联合全反式维甲酸对宫颈癌HeLa细胞株杀伤机制的研究

摘要

目的 探讨曲古菌素A(TSA)联合全反式维甲酸(ATRA)对宫颈癌HeLa细胞株的杀伤作用及其作用机制.方法 TSA联合 ATRA 或单独处理HeLa细胞,CellTiter 96 Aqueous 法检测细胞增殖抑制情况;7-AAD荧光染色法观察细胞凋亡改变;流式细胞术检测细胞凋亡率;Western blot 法检测p21、p53、Bcl-2、Caspase-3蛋白表达变化.结果 TSA联合ATRA对HeLa细胞具有明显的生长抑制作用,诱导细胞凋亡,出现核碎裂等典型凋亡改变,细胞凋亡率明显升高,与对照组和单独用药组比较,差异有统计学意义(P<0.05);与对照组比较,TSA联合ATRA组中p21、p53、活化性Caspase-3蛋白的表达显著升高(P<0.01),而Bcl-2蛋白表达显著下降(P<0.05).结论 TSA联合ATRA在体外对HeLa细胞有明显的杀伤作用,推测其机制可能是上调p21、p53、活化性Caspase-3蛋白表达水平,下调Bcl-2蛋白表达水平有关.%Objective To investigate the action mechanism of trichostatin A( TSA ) combined with all-trans retinoic acid( ATRA ) on HeLa cells. Methods HeLa cells were treated with TSA combined with ATRA or alone. CellTiter 96?AQueous assay was used to assess inhibitory rate of proliferation. The apoptosis of the cells were observed by 7-AAD fluorescent staining. The cell apoptosis rate were analyzed by flow cytometry. The protein expression levels of p21 ,p53,Bcl-2 and Caspase-3 were determined by Western blot. Results TSA combined with ATRA significantly inhibited the proliferation of HeLa cells, induced cell apoptosis and nuclear fragmentation. The apoptosis rate was significant rise, the difference was significant compared with control group and individual group( P < 0. 05 ). Compared with control group, protein expression levels of p21 ,p53 and active caspase-3 were significantly up-regulated ( P < 0. 01 ), but Bcl-2 was significantly down-regulated( P < 0. 05 ) in TSA combined ATRA group. Conclusion TSA alone or in combination with ATRA effectively kills cervical cancer HeLa cells in vitro. The effect may be related with up-regulation of p53 ,p21, active caspase-3 and down-regulation of Bcl-2.