microRNA-224对缺氧诱导结肠癌细胞凋亡的影响

Effect of microRNA-224 on apoptosis of colon cancer cells induced by hypoxia

目的:通过结肠癌细胞实验检测microRNA-224在结肠癌发生发展过程中的作用。方法:建立缺氧导致结肠癌细胞凋亡的模型,细胞计数盒(Cell Counting Kit-8,CCK-8)法检测结肠癌细胞活力;Annexin/PI流式细胞术检测结肠癌细胞凋亡率的变化。Annexin/PI流式细胞术检测转染microRNA-224后结肠癌细胞凋亡的变化。蛋白免疫印迹检测转染microRNA-224后凋亡蛋白Caspase-3的变化。结果:凋亡率在缺氧的结肠癌细胞组高于正常结肠癌细胞组(P<0.05);缺氧诱导凋亡组转染microRNA-224后凋亡率低于空质粒对照组(P<0.05);在缺氧诱导凋亡组中转染microRNA-224,凋亡相关蛋白Caspase-3的表达低于对照组(P<0.05)。结论:microRNA-224可抑制结肠癌细胞凋亡,结肠癌细胞的凋亡失衡在结肠癌的发生发展过程中具有重要意义。

Objective:To detect the role of microRNA-224 in the occurrence and development of colon cancer by colon cancer cell experiment.Methods:After the apoptosis model of colon cancer cells caused by hypoxia was established,we used Cell Counting Kit-8 to analyse the cell viability in hypoxia colon cancer cells group(hypoxia group)and normal colon cancer cells group(control group).Flow cytometry(FCM)was used to detect the rate of apoptosis in hypoxia colon cancer cells group(hypoxia group)and colon cancer cells group(control group),also detect the rate of apoptosis in microRNA-224-transfected colon cancer cells group(microRNA-224 group)and Vector-NC control group.Western blot was used to detect the proteins of apoptosis Caspase-3 in microRNA-224-transfected colon cancer cells group(microRNA-224 group)and Vector-NC control group.Results:After treatment with hypoxia in colon cancer cells,the apoptotic rate of cells was significantly higher than the control group(P<0.05).After pre-transfecting with microRNA-224,the apoptotic rate of cells was lower in hypoxia-treatment colon cancer cells than Vector-NC control group(P<0.05).The apoptotic protein Caspase-3 in microRNA-224-transfected colon cancer cells was lower than Vector-NC control group(P<0.05).Conclusion:MicroRNA-224 can inhibit the apoptosis of colon cancer cells,which has significant influence in the development of colon cancer.