沉默EZH2基因对人结肠癌LoVo细胞生物学功能的研究

Effect of silencing EZH2 gene on the biological function of human colon cancer LoVo cells

目的对人结肠癌LoVo细胞的果蝇zeste基因增强子同源物2(enhancer of zeste homo-log 2,EZH2)基因进行沉默并对其侵袭和转移功能研究,探索干扰EZH2是否是结肠癌细胞的治疗潜在靶点。方法设计并合成针对人EZH2基因的shRNA引物,并将其基因连接至pLVX-shRNA2质粒载体上,通过脂质体Lipodfectine2000转染人结肠癌LoVo细胞,通过免疫印迹鉴定EZH2的表达。并用Transwell方法研究干扰人EZH2基因对人结肠癌LoVo细胞的侵袭结果,细胞划痕实验研究干扰人EZH2基因对人结肠癌LoVo细胞的迁移结果。结果成功构建干扰人EZH2表达的p LVX-shRNA2质粒载体,有效抑制EZH2基因的表达。功能研究初步证明,干扰EZH2基因表达可抑制人结肠癌LoVo细胞的侵袭及细胞迁移。结论沉默EZH2基因显著下调人结肠癌LoVo细胞的侵袭和转移功能,可作为结肠癌治疗的潜在靶点之一。

Objective The human chromosome of zeste homo-log 2( EZH2) gene was silenced in human colon cancer LoVo cells,and its invasion and metastasis function were studied,so as to explore whether EZH2 interference can be a potential target for colon cancer treatment. Methods The shRNA primer against human EZH2 gene was designed and synthesized,and its gene was ligated into p LVX-shRNA2 plasmid vector. Human colon cancer LoVo cells were transfected by lipofect Lipodfectine2000,and the expression of EZH2 was identified by immunoblotting. Transwell method was used to study the invasion of human colon cancer LoVo cells by interfering with human EZH2 gene. Cell scratch assay was used to investigate the migration of human EZH2 gene to human colon cancer LoVo cells. Results The p LVX-shRNA2 plasmid vector which interfered with the expression of human EZH2 was successfully constructed,and the expression of EZH2 gene was effectively inhibited. Functional studies initially demonstrated that interference with EZH2 gene expression inhibited invasion and cell migration of human colon cancer LoVo cells. Conclusion Silencing EZH2 gene significantly down-regulates the invasion and metastasis of human colon cancer LoVo cells,which may be one of the potential targets for colon cancer therapy.