RNAi沉默clusterin基因对卵巢癌SKOV3细胞放射敏感性的影响

Influence of silencing clusterin by RNAi on radiosensitivity of human ovarian carcinoma SKOV3 cell-line

目的:研究clusterin(CLU)基因沉默对卵巢癌SKOV3细胞放射敏感性的影响。方法:靶向CLU基因的siRNA转染卵巢癌SKOV3细胞,Western blot法测定CLU基因蛋白表达水平。给予不同放射剂量(2、4、6 Gy)处理后,通过MTT法及克隆形成实验检测卵巢癌SKOV3细胞的存活率及克隆形成率,流式细胞仪Annexin V/PI法检测凋亡率。结果:靶向CLU基因的siRNA特异性地从蛋白水平抑制了CLU基因表达。MTT及克隆形成实验结果显示卵巢癌SKOV3细胞的存活率及克隆形成率与放射剂量呈反比,剂量越大,存活率及克隆形成率越低,差异均具有统计学意义(P<0.05)。流式细胞仪Annexin V/PI法检测结果表明:2 Gy放疗后实验组细胞凋亡率明显增加,达(24.32±1.75)%,较单纯放射组升高约12%,差异具有明显统计学意义(P<0.05)。结论:CLU基因沉默可以明显增强卵巢癌SKOV3细胞对放射的敏感性,为卵巢癌放疗增敏基础研究提供了新的靶点。

Objective：To investigate the influence of silencing clusterin（CLU） by RNA interference on radiosensitivity of human ovarian carcinoma SKOV3 cell-line. Methods：The expression of CLU gene was silenced by siRNA against it, Western blot was performed to detect the expression of C LU on protein level. After receiving different doses of radiation（2,4,6 Gy）, The cell survival rate was detected by MTT assay,and the cloning efficiency was also evaluated by the assay,the apoptotic rate was detected by flow cytometry. Results：The siRNA target clu specifically down regulated the expression of CLU significantly. The MTT and colony forming results showed that the cell survival and cloning efficiency became significantly lower with the increasing dose of ionizing irradiation, the capacity of proliferation and cloning efficiency was inhibited obviously（P〈0.05）. The apoptotic rate became significantly higher with the increasing dose of ionizing irradiation,which reached（24.32±1.75）% after 2 Gy（P〈0.05）, and increased about 12% compared with control group. Conclusion：The CLU-silencing can significantly enhance the radio-sensitivity of ovarian carcinoma SKOV3 cellline in vitro, and the CLU gene may be a promising target for basic research of ovarian carcinoma. [