线粒体途径在温热化疗诱导胃癌细胞AGS凋亡中的作用

目的探讨线粒体途径在温热化疗诱导胃癌细胞AGS凋亡中的作用。方法应用人胃癌细胞株AGS,根据不同实验处理分为常温对照(NT)、单纯温热(HT)、常温化疗(NTCT)和温热化疗(HTCT)四组。Western blotting法检测AGS细胞中Bax、Bcl-2蛋白及其线粒体胞质蛋白中细胞色素C的表达,实时PCR检测其Bax、Bcl-2的mRNA表达;分别测定各组AGS细胞Caspase3和Caspase9的酶活性变化。结果与NT、HT、NTCT组比较,HTCT组AGS细胞Bcl-2蛋白表达下调而Bax蛋白表达上调,Bax/Bcl-2显著上升;线粒体胞质蛋白中细胞色素C表达增加;Caspase9和Caspase3的酶活性显著增高。结论HTCT可引起AGS细胞Bcl-2、Bax基因和蛋白表达水平发生改变,促使其细胞色素C由线粒体释放进入细胞质,从而诱导细胞凋亡。

Role of mitochondria pathway in apoptosis induction of gastric cancer cell line AGS under hyperthermic chemotherapy

Objective To evaluate the role of mitochondria pathway in the apoptosis induction of gastric cancer cell line AGS under hyperthermic chemotherapy(HTCT).Methods Human gastric cancer cell line AGS was used in this study.Treatment conditions were categorized into 4 modes:normothermic control(NT),hyperthermia(HT),normothermic chemotherapy(NTCT)and HTCT.The expressions of Bax,Bcl-2 and cytochrome C in gastric cancer cell line AGS were determined by Western blotting,and the expressions of Bax and Bcl-2 mRNA were determined by real-time PCR.The changes for enzyme activity of Caspase 3 and Caspase 9 were also detected.Results Compared with the NT,HT and NTCT modes,under HTCT,the expression of Bax protein in AGS cells was up-regulated and the expression of Bcl-2 protein was down-regulated,resulting in the significant increase of Bax/Bcl-2 ratio.Cytochrome C was released from mitochondria to cytosol,with a significant increase of enzyme activities of Caspase 9 and Caspase 3.Conclusion HTCT may lead to the change of Bax and Bcl-2 mRNA and protein experessions in gastric cancer cell line AGS,facilitate the release of cytochrome C from mitochondria to cytosol and induce the cell apoptosis.