PES1基因小干扰RNA慢病毒载体的构建及其对乳腺癌ZR75-30细胞的生长抑制

目的:构建PES1基因的慢病毒干扰载体,研究其对乳腺癌细胞ZR75-30生长的影响。方法:设计针对PES1基因的siRNA引物,克隆到pSIH1-H1-Puro载体,包装成慢病毒,测定病毒的滴度,感染人乳腺癌细胞ZR75-30,通过Real-time PCR和Western印迹检测干扰效果,并通过生长曲线研究敲低PES1对ZR75-30细胞生长的影响。结果和结论:构建了具有明显干扰效果的PES1基因的siRNA慢病毒载体,能够有效抑制ZR75-30细胞内源的PES1 mRNA和蛋白水平,敲低PES1能够抑制ZR75-30细胞的生长。

Construction of a Lentiviral Vector for PES1 Gene Small-Interfering RNA and its Inhibition on the Growth of ZR75-30 Breast Cancer Cells

Objective：To construct a lentiviral vector for PES1 small-interfering RNA（siRNA）,and to study its effect on the growth of ZR75-30 breast cancer cells.Methods：siRNA primers targeting PES1 were designed and cloned into the vector pSIH1-H1-Puro.Lentivirus was obtained and the titer was tested.Real-time PCR and Western blot were performed to identify the lentivirus-mediated knockdown of PES1.Growth curve was performedto study the effect of PES1 knockdown on the growth of ZR75-30.Results Conclusion：A lentivirus RNA in-terference vector targeting PES1 gene was sucessfully constructed.Knocking down PES1 could repress the growth of ZR75-30 cells.