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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 role of prohibitin (PHB) gene in inducing gastric cancer BGC823 cell apoptosis. Methods: BGC823 cells with low and high expression of PHB gene were obtained by RNA interference and gene transfection respectively. The proliferation, cell cycle and apoptosis of PHB gene were detected by MTT assay and FCM respectively. The expression of Bax, Bcl-2 and cytoplasmic cytochrome c were detected by Western blot, and The activation of caspase-3 and caspase-9 were detected. Results: After transfection of PHB gene into BGC823 gastric cancer cells, the cell growth rate slowed down, the proportion of cells in G1 phase decreased significantly, the proportion of cells in G2 phase increased slightly, the proportion of S phase cells and apoptosis rate increased significantly, Bax increased, Bcl -2 was down-regulated, cytochrome c levels in cytoplasm increased significantly, and the activation of caspase-3 and caspase-9 increased. PHB gene-specific RNA interference of gastric cancer cells BGC823 accelerated cell growth, G1 and G2 phase cells accounted for little change in the proportion of S phase cells increased slightly, the apoptosis rate decreased slightly, Bax expression down, Bcl-2 The expression of caspase-3 and caspase-9 activation was weakened. Conclusion: PHB is one of the important factors inhibiting the growth of gastric cancer cells and inducing apoptosis. It is speculated that this effect may be achieved through the mitochondrial pathway.