导入靶向survivin基因的shRNA质粒表达载体对大肠癌细胞生物学行为的影响

Biological behavior of colorectal carcinoma Lovo cells after stable transfection survivin targeted shRNA plasmid expression vector

目的:合成靶向survivin基因的shRNA并构建pGenesil-1-shRNA质粒表达载体,研究其对大肠癌Lovo细胞生物学效应的影响。方法:设计2条特异性靶向survivin基因的shRNA,与pGenesil-1质粒载体连接,导入大肠癌Lovo细胞、筛选稳定细胞株,Western Blot检测蛋白表达,流式细胞技术检测细胞生长周期,MTT法描绘细胞生长曲线。结果:筛选的稳定表达细胞内survivin蛋白的表达水平降低,survivin基因低表达的Lovo细胞停滞于G2/M期从而导致细胞生长速度减慢。结论:靶向survivin基因的shRNA能有效降低survivin基因在体内的表达,抑制大肠癌细胞的生长,为大肠癌基因治疗的可行性提供了有力的证据。

Objective To synthetize survivin targeted short hairpin RNA and constructed pGenesil-1-shRNA plasmids expression vectors and study the influence of the shRNA on the biological behavior of colorectal carcinoma Lovo cells.Methods Two pairs of shRNA sequences of target survivin gene were designed,and respectively inserted into pGenesil-1 plasmids vector.After stable transfection into Lovo cells,protein expression level was detected with Western Blot,cell growth cycle was detected with flow cytometry and cell growth curve was described with MTT assay. Results Survivin protein of two groups of stable expression cells were both much lower than normal Lovo cells, and almost stopped in stage G2/M and the growth velocity were turned to slow. Conclusion Survivin targeted shRNA could specifically suppress survivin expression in Lovo cells and inhibit colorectal carcinoma cells growth. It provides a powerful evidence of colorectal carcinoma gene therapy.