下调Drosha基因表达促进MGC-803胃癌细胞对表阿霉素的敏感性  

Knockdown of Drosha promotes chemosensitivity of epirubicin for gastric cancer MGC-803 cells

作　　者：徐丽云[1] 陈燕林[1] 文思阳 杜燕娥[1] 唐曦[1] 柳满然[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《细胞与分子免疫学杂志》2016年第9期1207-1211,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81172296)

摘　　要：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 establish a gastric cancer cell line with stable Drosha silenced and explore the effect of Drosha on the chemosensitivity of gastric cancer cells to epirubicin. Methods Interfering sequences targeting Drosha were designed and inserted into the lentiviral vectors, which were used to transfect MGC-803 cells. The level of Drosha mRNA was detected by quantitative real-time PCR; Drosha protein was detected by Western blotting; MTT assay was performed to test the 50% inhibitory concentration （ IC50 ） of epirubicin agaisnt wide-type MGC-803 cells. After the treatment with IC50 epirubicin, the apoptosis rate of each cell group was determined by flow cytometry; the expressions of apoptosis-related proteins caspase-3, caspase-9, Bax, Bcl-2 were assessed by Western blotting. Results The gastric cancer MGC-803 cells with stable Drosha silenced were successfully established, and the levels of Drosha mRNA and protein were reduced. After the cells were treated with 0.5 mg/L（ IC50 ）epirubicin, the apoptosis rate of MGC-803 cells was raised, the protein expressions of caspase-3, caspase-9 and Bax were significantly upregulated and Bcl-2 was downregulated. Conclusion The silence of Drosha expression can promote the sensitivity of gastric cancer to epirubicin.

关 键 词：DROSHA MGC-803胃癌细胞 表阿霉素 

分 类 号：R735[医药卫生—肿瘤] R392-33[医药卫生—临床医学]