bcl-3基因通过cyclin D1及Bax蛋白影响人结肠癌RKO细胞的凋亡

Effects of bcl-3 gene silencing on apoptosis of human colon cancer cell line RKO

目的:研究bcl-3基因对人结肠癌RKO细胞迁移及凋亡的影响及机制。方法:采用人bcl-3基因的RNA干扰慢病毒载体沉默人结肠癌RKO细胞bcl-3基因的表达后,划痕实验观察bcl-3基因沉默前后RKO细胞迁移能力的变化,Annexin V/PI双染色法检测bcl-3基因沉默前后RKO细胞凋亡率的变化,Western blot法检测bcl-3基因沉默前后细胞周期蛋白cyclin D1及凋亡相关蛋白Bax、Bcl-2的变化。结果:划痕实验显示,划痕后36 h,bcl-3基因沉默前后RKO细胞划痕愈合率分别为84.00%及40.00%,差异具有统计学意义(P<0.05)。Annexin V/PI双染色法流式细胞术分析显示,bcl-3基因沉默前后的RKO细胞均经5μmol/L顺铂处理24 h后,沉默前后的RKO细胞凋亡率分别为12.89%及59.67%,差异具有统计学意义(P<0.05)。Western blot法检测显示bcl-3基因沉默后cyclin D1蛋白表达显著下降(P<0.05),Bax蛋白表达显著上升(P<0.05),但Bcl-2表达无明显变化(P>0.05)。结论:沉默bcl-3基因后,RKO细胞迁移能力下降,凋亡率增加,并伴细胞周期蛋白cyclin D1及凋亡相关蛋白Bax表达的变化。bcl-3基因可能通过改变cyclin D1及Bax蛋白的表达而影响RKO细胞的凋亡。

AIM： To investigate the effects of bcl-3 gene on the migration and apoptosis of human colon cancer cell line RKO, and the changes of cyclin D1 and apoptosis-related proteins.METHODS： After silencing of bcl-3 gene expression in human colon cancer cell line RKO by lentiviral vector with RNA interference, the changes of RKO cell migration ability were investigated by wound healing assay. The changes of RKO cell apoptotic rate after bcl-3 gene silencing were detected by flow cytometry with Annexin V/PI staining. The protein expression of cyclin D1 and apoptosis-related proteins in the RKO cells after bcl-3 gene silencing was determined by Western blot.RESULTS： The wound healing rates of the RKO cells were 84.00% and 40.00% before and after bcl-3 gene silencing, respectively, with statistically significant difference （P〈0.05）. Annexin V/PI staining showed that the cell apoptotic rates were 12.89% and 59.67% before and after bcl-3 gene silencing, respectively, when RKO cells were treated with 5 μmol/L cisplatin for 24 h, with statistically significant difference （P〈0.05）. The expression of cyclin D1 decreased, while the expression of Bax increased after bcl-3 gene silencing （P〈0.05）.CONCLUSION： After bcl-3 gene silencing, the migration ability of RKO cells decreases, and the apoptotic rate increases, accompanying with the changes of cyclin D1 and Bax. bcl-3 gene can affect the apoptosis of RKO cells by changing the expression of cyclin D1 and Bax.