p21基因沉默联合表阿霉素促进乳腺癌MCF-7细胞凋亡及其机制的研究

Si RNA inhibition of p21 combined with Epirubicin induced apoptosis of MCF-7 cells

目的研究si RNA沉默p21基因表达联合表阿霉素对MCF-7细胞增殖和凋亡的影响及相关作用机制。方法将化学合成的针对p21的si RNA序列转染至MCF-7细胞,将MCF-7细胞分为5组:空白对照组、阴性对照组、RNA抑制组、表阿霉素组、联合组(RNA沉默+表阿霉素)。采用MTT比色法、流式细胞术和分光光度法分别检测MCF-7细胞增殖、细胞凋亡、细胞周期及半胱氨酸天冬氨酸蛋白酶家族成员Caspase-9、Caspase-3和Caspase-6的活化程度。结果 p21 si RNA转染后,相对于表阿霉素单独处理组,联合组细胞死亡率明显升高(P<0.01);Caspase-9、Caspase-3和Caspase-6的活化程度显著升高(P<0.01)。结论 p21靶向RNA抑制联合表阿霉素处理显著促进了MCF-7细胞凋亡,p21可作为乳腺癌基因治疗的后选新靶点。

Objective To study the influence of si RNA inhibition of p21 combined with EPI treatment on proliferation and apoptosis of MCF-7 cells and its related mechanism.Methods The si RNA sequence targeting p21 which was synthesized by chemical method was transfected into MCF-7 cells.The MCF-7 cells were divided into five groups： blank control group,negative control group,si RNA group,EPI treated group and combination group（si RNA combined with EPI treatment）.MTT assay,flow cytometry and spectrophotometry were employed to detect the variation of cell viability,apoptosis and Caspase-9,Caspase-3 and Caspase-6 activity.Results After transfection,compared with the EPI treated group,the cell death rate of the combination group were markedly increased（P〈0.01）,and activity of Caspase-9,Caspase-3 and Caspase-6 was obviously in-duced（P〈0.01）.Conclusion Si RNA combined with EPI treatment can significantly induce cell apoptosis,p21 gene may be a candidate target in the gene therapy of breast cancer.