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目的使用针对E盒锌指结合蛋白1(ZEB1)的siRNA在结肠癌HCT116细胞中下调ZEB1的表达,研究其对HCT116细胞增殖和凋亡的影响。方法根据人ZEB1的mRNA序列设计并合成2对针对ZEB1的siRNA,瞬时转染HCT116细胞后,利用实时荧光定量PCR,Western blot法分别在mRNA和蛋白水平检测siRNA对ZEB1的抑制效果;利用MTT法检测细胞增殖;AnnexinⅤ-FITC/PI染色结合流式细胞术检测细胞凋亡情况。结果与对照组相比,针对ZEB1的siRNA可在mRNA和蛋白水平有效抑制ZEB1的表达,ZEB1的表达下调可有效抑制HCT116细胞的增殖,并促进细胞凋亡。结论 ZEB1对结肠癌细胞的生存和增殖具有重要作用,可能参与结肠癌的发生发展过程。
OBJECTIVE: To investigate the effect of ZEB1 on the proliferation and apoptosis of HCT116 cells by siRNA targeting E box zinc finger binding protein 1 (ZEB1) in human colon cancer HCT116 cells. Methods Two pairs of siRNA targeting ZEB1 were designed and synthesized based on human ZEB1 mRNA sequence. After transient transfection of HCT116 cells, the inhibitory effect of siRNA on ZEB1 was detected by real-time fluorescence quantitative PCR and Western blot respectively. Cell proliferation was detected by flow cytometry. AnnexinⅤ-FITC / PI staining and flow cytometry were used to detect cell apoptosis. Results Compared with the control group, siRNA targeting ZEB1 could effectively inhibit the expression of ZEB1 at mRNA and protein level. The down-regulation of ZEB1 could inhibit the proliferation and promote the apoptosis of HCT116 cells. Conclusion ZEB1 plays an important role in the survival and proliferation of colon cancer cells and may be involved in the development and progression of colon cancer.