RNA干扰靶向抑制PI3Kp85α蛋白对大肠癌细胞增值的影响

The Influence of Silencing PI3K p85α Expression by RNA Interference on Proliferation of Colorectal Cancer Cells

目的:探讨RNA干扰靶向抑制PI3K p85α蛋白表达对大肠癌细胞增值的影响。方法:设计4条shRNA干扰载体及1条阴性对照载体,分别稳定转染大肠癌SW480细胞。用Western Blot筛选出一组抑制效率最高的细胞进行CCK-8细胞增值实验。结果:转染shRNA/324干扰载体的SW480细胞PI3K p85蛋白表达显著降低,抑制率约90%,选择该组细胞作为有效干扰组。CCK-8细胞增值实验显示,接种48h、72h及96h后,干扰组细胞生长速度明显低于对照组细胞(P<0.05)。结论:PI3K p85α蛋白表达缺失可降低大肠癌SW480细胞的增值速度,PI3K p85可能是大肠癌靶向治疗的新靶点。

Objective： To investigate the effect of PI3K p85α depletion via RNA interference on proliferation of colorectal cancer cells. Methods： Four shRNA vectors and one negative control vector were designed and stably transfected into SW480 cells. Western Blot analysis was used to determine the RNA interference effect of expression level of PI3K p85α protein. Cell proliferation was determined using Cell Counting Kit-8 （CCK-8） solution after stable transfection. Results. Western Blot analysis showed that the expression level of PI3K p85α protein was significantly decresed in cells transfected with shRNA/324 vector, the inhibited rate was 90%, the group was selected for the following experiments. Compared with the control cells, SW480 shRNA/324 cells grew much slowly at 48h, 72h and 96h （P〈0.05）. Conclusions. Depletin of PI3K p85α could significantly inhibit proliferation of SW480 cells. PI3K p85α may be a new therapeutic target for colorectal cancer cells.