RNAi沉默β-arrestin2基因对人乳腺癌Bcap37细胞凋亡的影响

摘要

Objective:To observe apoptosis of human breast cancer Bcap37 cells by siRNA of β-arrestin2 in vitro.Methods:The vector PGPU6/GFP/Neo cloned β-arrestin2 small interfering RNA(siRNA)was transformed in-to human breast cancer Bcap37 cells.The transfected PGPU6/GFP/NC was used as a negative control and the non-transfected vector served as a blank control.The expression of β-arrestin2 gene was monitored by Western blot.Cell apoptosis was examined by Flow cytometry.Results:Our study showed that β-arrestin2-siRNA efficiently downreg-ulated the expression of β-arrestin2 in β-arrestin2-siRNA group compared with the negative and blank control(P<0.05).Bcap37 cells in low expression of β-arrestin2 caused by siRNA represented an upward trend of apoptosis, but not in the negative and blank controls.Conclusion:Our results suggested that β-arrestin2-siRNA can effective-ly inhibit the expression of β-arrestin2,promote the apoptosis of human breast cancer Bcap37 cells.These results demonstrate that suppression of β-arrestin2 activity by siRNA may be an effective strategy for gene therapy in human breast cancers.%目的:通过体外实验观察β-arrestin2的小分子干扰RNA(small interfering RNA,siRNA)对人乳腺癌Bcap37细胞凋亡的影响.方法:针对β-arrestin2基因设计siRNA序列,克隆到空载体PGPU6/GFP/Neo,转化DH5α菌株,提取质粒,进行测序分析.在脂质体lipofectamineTM2000的介导下转染Bcap37细胞,同时转染空白对照组及阴性对照组.转染后用定量Western blot检测Bcap37细胞β-arrestin2蛋白的表达情况,流式细胞仪检测Bcap37细胞的凋亡情况.结果:我们的研究表明,β-arrestin2的siRNA能有效下调β-arrestin2的表达,与对照组相比,β-arrestin2干扰组 β-arrestin2的表达明显下降(P <0.05);β -arrestin2干扰组Bcap37细胞凋亡率显著高于对照组.结论:β-arrestin2干扰组能有效降低β-arrestin2的表达,有效促进Bcap37细胞凋亡;β-arrestin2可能成为乳腺癌基因治疗的一个新靶点.