论文部分内容阅读
目的构建Drosha基因稳定沉默的胃癌细胞株并探讨其对表阿霉素敏感性的影响。方法将靶向干扰Drosha的序列重组到慢病毒载体并感染MGC-803细胞;采用实时定量PCR检测Drosha mRNA水平、Western blot法检测Drosha蛋白水平;MTT法检测野生型MGC-803细胞对表阿霉素的半数抑制剂量(IC50);流式细胞术检测用IC50的表阿霉素处理后各组细胞的凋亡率,Western blot法检测凋亡相关蛋白caspase-3、caspase-9、Bax、Bcl-2的蛋白水平。结果成功构建稳定Drosha基因沉默的MGC-803细胞株;下调Drosha后,经0.5 mg/L(IC50)表阿霉素处理,MGC-803细胞凋亡率明显增加,MGC-803细胞caspase-3、caspase-9、Bax表达上调,Bcl-2表达降低。结论沉默细胞中Drosha表达能提高胃癌细胞对表阿霉素的敏感性。
Objective To construct a stable gastric cancer cell line with Drosha gene and investigate its effect on epirubicin sensitivity. Methods The Drosha target sequence was subcloned into lentiviral vector and infected MGC-803 cells. The Drosha mRNA level was detected by real-time quantitative PCR and the Drosha protein level was detected by Western blot. The effect of wild-type MGC-803 (IC50). Flow cytometry was used to detect the apoptotic rate of each group after treated with epirubicin with IC50. Western blot was used to detect the expressions of caspase-3, caspase-9, Bax and Bcl- -2 protein levels. Results The MGC-803 cell line with stable Drosha gene silencing was constructed successfully. After Drosha down-regulation, the apoptotic rate of MGC-803 cells was significantly increased after treated with epirubicin at 0.5 mg / L (IC50). The caspase-3, caspase-9, Bax expression is up-regulated, Bcl-2 expression is reduced. Conclusion Drosha expression in silencing cells can enhance the sensitivity of gastric cancer cells to epirubicin.