阻抑素基因在诱导胃癌BGC823细胞凋亡过程中的作用研究  

The role of prohibitin gene in inducing apoptosis of gastric cancer BGC823 cells

作　　者：张隆[1] 季青[1] 吴蓉[2] 孙健[1] 刘丽丽[2] 康向东[2] 

机构地区：[1]上海中医药大学附属普陀医院实验中心,上海200062 [2]上海中医药大学附属普陀医院检验科,上海200062

出　　处：《肿瘤》2010年第11期929-933,共5页Tumor

基　　金：上海市教育委员会基金资助项目(编号:07cz016)

摘　　要：目的:研究阻抑素(prohibitin,PHB)基因在诱导胃癌BGC823细胞凋亡过程中的作用。方法:使用RNA干扰技术和基因转染技术,分别获得PHB基因低表达和高表达的胃癌BGC823细胞。然后采用MTT法和FCM法分别检测PHB基因低表达和高表达的胃癌细胞的增殖率、细胞周期和凋亡情况,Western印迹法检测Bax、Bcl-2和细胞质内细胞色素c的表达水平,并且检测caspase-3和caspase-9的激活情况。结果:PHB基因转染胃癌BGC823细胞后,细胞生长速度减缓,G1期细胞所占比例明显下降,G2期细胞比例略有上升,S期细胞比例和细胞凋亡率明显上升,Bax表达上调,Bcl-2表达下调,细胞质内细胞色素c水平显著上升,caspase-3和caspase-9的激活程度增加。PHB基因特异性RNA干扰胃癌细胞BGC823后,细胞生长加速,G1期和G2期细胞所占比例变化不大,S期细胞比例略有上升,细胞凋亡率小幅下降,Bax表达下调,Bcl-2表达上调,caspase-3和caspase-9的激活程度减弱。结论:PHB是抑制胃癌细胞生长和诱导细胞凋亡的重要因子之一,推测该作用可能是通过线粒体途径实现的。Objective:To study the function of prohibitin（PHB） gene in inducing apoptosis of gastric cancer BGC823 cells.Methods:PHB gene was silenced using RNA interference technology and was transfected using gene transfer techniques to obtain the two cell lines with the low or high expression of PHB,respectively.The cell proliferation was measured using MTT assay.Cell cycle and apoptosis were determined using flow cytometry.Bax,Bcl-2,and cytochrome c protein expressions and the activations of caspase-3 and caspase-9 were detected using Western blotting.Results:When the PHB gene was transfected into the BGC823 cells,the cells grew slowly;The proportion of cells in G1 phase decreased significantly,and the proportion of cells in G2 phase increased slightly;The proportion of cells in S phase was significantly elevated,and the apoptotic rate was significantly increased;The Bax protein and cytochrome c protein expressions were up-regulated,the expression of Bcl-2 protein was down-regulated;the activation levels of caspase-3 and caspase-9 were increased.When the PHB gene was silenced by RNA interference in BGC823 cells,the cells grew quickly;the proportion of cells in G1 and G2 phase had little changes;the proportion of cells in S phase increased slightly;the rate of apoptosis slightly decreased;the Bax protein expression was down-regulated and Bcl-2 expression was up-regulated;the activation levels of caspase-3 and caspase-9 were decreased.Conclusion:PHB was one of the important factors in inhibiting growth and inducing apoptosis of gastric cancer cells.The effects may be exerted through mitochondrial pathway.

关 键 词：基因低表达 诱导细胞凋亡 胃癌细胞 BGC823细胞 凋亡过程 cells gastric cancer caspase-9 caspase-3 癌细胞生长 细胞凋亡率 PHB Bcl-2表达 细胞质内 细胞色素 基因转染 RNA干扰技术 BAX表达 WESTERN印迹法 检测 

分 类 号：R73[医药卫生—肿瘤]