C-myc-siRNA通过调控ROS诱导子宫内膜癌细胞凋亡

C-myc-siRNA could Induce Apoptosis of Endometrial Carcinoma Cell Through Regulating Reactive Oxygen Species

为探讨原癌基因C-myc-siRNA对子宫内膜癌细胞凋亡的影响,本研究利用细胞计数盒(cell counting Kit-8, CCK-8)法分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞活力;蛋白免疫印迹法(Western blotting)分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞凋亡蛋白Bax以及抗凋亡蛋白Bcl-2的变化;流式细胞仪分别检测C-myc-siRNA转染组和空载体转染组(Vector-NC)活性氧簇(reactive oxygen species, ROS)的变化;蛋白免疫印迹法(Western blotting)分别检测预处理NAC (ROS抑制剂)后,C-myc-siRNA转染组和空载体转染组(Vector-NC)的子宫内膜癌细胞凋亡相关蛋白Bax以及Bcl-2的变化。结果表明:C-myc-siRNA转染子宫内膜癌细胞后,促凋亡相关蛋白Bax的表达显著高于空载体转染组,且抗凋亡相关蛋白Bcl-2的表达明显低于空载体转染组(p<0.05);C-myc-siRNA转染子宫内膜癌细胞后,细胞ROS水平明显增加(p<0.05);NAC预处理显著减弱C-myc-siRNA对子宫内膜癌细胞凋亡的促进作用(p<0.05)。本研究结论表明,C-myc-siRNA能够通过调控ROS诱导子宫内膜癌细胞凋亡。

In order to discuss the effects of C-myc-siRNA on the apoptosis of endometrial carcinoma cell.In this research,Cell Counting Kit-8 was used to detect endometrial carcinoma cell viability of C-myc-siRNA-transfected group and Vector-NC group.Western blotting was used to detect the changes of endometrial carcinoma apoptosis protein Bax and anti-apoptosis protein Bcl-2 in C-myc-siRNA transfected group and Vector-NC group.Flow cytometry(FCM)was used to evaluate the changes of reactive oxygen species(ROS)in C-myc-siRNA transfected group and Vector-NC group.Western blot was used to detect the changes of endometrial carcinoma apoptosis related protein Bax and Bcl-2 in C-myc-siRNA transfected group and Vector-NC group after the pre-treatment with NAC(ROS inhibitor).The results revealed that after C-myc-siRNA transfected endometrial carcinoma cell,the expression of pro-apoptotic related protein Bax was significantly higher than that in Vector-NC group and the expression of anti-apoptotic related protein Bcl-2 was significantly lower than that in Vector-NC group(p<0.05).After C-myc-siRNA transfected endometrial carcinoma cell,the ROS level of endometrial carcinoma cell was significantly higher(p<0.05).Pretreatment with NAC significantly weakened the acceleration of C-myc-siRNA on the apoptosis of endometrial carcinoma cell(p<0.05).The conclusion of this study demonstrated that C-myc-siRNA could induce the apoptosis of endometrial carcinoma cell by the regulation of reactive oxygen species.