EIF3D shRNA对乳腺癌细胞增殖、凋亡与侵袭能力的影响

目的探讨真核起始因子3D(EIF3D)shRNA对乳腺癌细胞增殖、凋亡与侵袭能力的影响。方法设计靶向EIF3D的shRNA序列,构建稳定沉默EIF3D的慢病毒载体,感染乳腺癌细胞MCF-7,同时设立阴性对照组与未处理组。qPCR和Western blot方法测定沉默效果,MTT方法测定感染后MCF-7细胞增殖能力变化,流式细胞术测定感染后MCF-7细胞凋亡率,Transwell侵袭实验检测感染后MCF-7细胞侵袭能力变化,Western blot方法测定感染后MCF-7细胞中Cyclin B1、基质金属蛋白酶-2(MMP-2)与Caspase-3的表达变化。结果感染EIF3D shRNA后,MCF-7细胞EIF3D的蛋白与mRNA表达均显著降低,提示稳定沉默EIF3D细胞株模型成功建立。沉默EIF3D能够显著抑制MCF-7细胞的增殖与侵袭能力,诱导细胞凋亡,下调Cyclin B1与MMP-2的蛋白表达,上调Caspase-3的表达(P<0.01)。结论下调EIF3D能够抑制乳腺癌MCF-7细胞增殖与侵袭能力,并诱导细胞凋亡。

Effects of EIF3D shRNA on proliferation,apoptosis and invasion of breast cancer cells

Objective To investigate the effect of eukaryotic translation initiation factor 3,subunit D(EIF3 D)shRNA on the proliferation,apoptosis and invasion of breast cancer cells.Methods The shRNA sequence targeting EIF3 D was designed to construct a stable EIF3 D silenced lentivirus vector and infect breast cancer cell MCF-7.At the same time,negative control group and untreated group were set up.MTT method was used to detect the proliferation of MCF-7 cells,flow cytometry was used to detect the apoptosis rate of MCF-7 cells,Transwell assay was used to detect the invasion ability of MCF-7 cells.The expression of cyclin B1,matrix metalloproteinase-2(MMP-2)and Caspase-3 in MCF-7 cells after transfection were determined by Western blot.Results After EIF3 D shRNA was transfected,the expression of EIF3 D protein and mRNA in MCF-7 cells was decreased significantly,and the cell line model of stable silencing EIF3 D was successfully established.Silencing EIF3 D could significantly inhibit the proliferation and invasion of MCF-7 cells,induce apoptosis,down regulate the expression of cyclin B1 and MMP-2,and up regulate the expression of Caspase-3(P<0.01).Conclusion Down regulation of EIF3 D could inhibit the proliferation and invasion of breast cancer MCF-7 cells,and induce apoptosis.