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目的观察凋亡抑制蛋白Survivin基因干扰和过表达对人乳腺癌MCF-7细胞凋亡的影响。方法将Survivin基因shRNA干扰质粒和过表达质粒pcDNA3.1-GFP-Survivin在脂质体介导下转染MCF-7细胞,并设空白对照组和脂质体对照组,转染后48 h,荧光显微镜下观察转染效率;荧光定量RT-PCR法检测转染细胞中Survivin基因mRNA的转录水平;MTT法检测转染细胞的凋亡率。结果转染MCF-7细胞后48 h,Survivin基因RNAi质粒和过表达质粒的转染效率为50%～60%;shRNA质粒转染组MCF-7细胞中Survivin基因mRNA的转录水平明显低于空白对照组和脂质体对照组,而过表达质粒转染组明显高于空白对照组和脂质体对照组(P<0.05);shRNA质粒转染组MCF-7细胞的凋亡率明显高于空白对照组和脂质体对照组,而过表达质粒转染组明显低于空白对照组和脂质体对照组(P<0.05)。结论 Survivin基因对人乳腺癌MCF-7细胞的凋亡具有一定的抑制作用,可作为人乳腺癌生物治疗的候选基因。
Objective To observe the effect of Survivin gene interference and overexpression on the apoptosis of human breast cancer MCF-7 cells. Methods shRNA interference plasmid of Survivin gene and pcDNA3.1-GFP-Survivin plasmid were transfected into MCF-7 cells by lipofectamine 2000. Blank control group and liposome control group were established. After 48 h of transfection, The transfection efficiency was observed under a fluorescence microscope. The transcription level of Survivin mRNA in transfected cells was detected by fluorescence quantitative RT-PCR. The apoptosis rate of transfected cells was detected by MTT assay. Results The transfection efficiency of Survivin RNAi plasmid and overexpression plasmid was 50% -60% at 48 h after transfection of MCF-7 cells. The transcription level of Survivin mRNA in MCF-7 cells transfected with shRNA plasmid was significantly lower than that of blank (P <0.05). The apoptosis rate of MCF-7 cells transfected with shRNA plasmid was significantly higher than that of the control group and the liposome control group, while the transfection group with the overexpression plasmid was significantly higher than the blank control group and the liposome control group The blank control group and the liposome control group, while the overexpression plasmid transfected group was significantly lower than the blank control group and the liposome control group (P <0.05). Conclusion Survivin gene can inhibit the apoptosis of human breast cancer cell line MCF-7 and may serve as a candidate gene for human breast cancer biotherapy.