RNA干扰靶向抑制PI3K p85α蛋白对大肠癌细胞侵袭与转移的影响

Influence of RNA interference targeting PI3K p85α expression on invasion and metastasis of colorectal cancer cells

目的:探讨RNA干扰靶向抑制PI3Kp85α蛋白表达对大肠癌细胞侵袭与转移的影响。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LoVo细胞。用蛋白质印迹法筛选出一组抑制效率最高的细胞进行细胞划痕实验和Transwell实验。结果:转染shRNA/324干扰载体的LoVo细胞PI3Kp85α蛋白表达降低最明显,抑制率达77%,选择该组细胞作为有效干扰组进行后续实验。细胞划痕实验显示,干扰组细胞的划痕愈合能力明显低于对照组细胞(P<0.05)。Transwell实验显示,干扰组细胞的穿膜细胞数明显少于对照组细胞(P<0.05)。结论:PI3Kp85α蛋白表达缺失可明显降低大肠癌LoVo细胞的迁移运动能力,PI3Kp85α可能成为治疗大肠癌转移的新靶点。

Objective：To investigate the influence of RNA interference targeting PI3K p85α on the invasion and metastasis of coloreetal cancer cells. Methods： Four shRNA vectors and one negative control vector were designed and stably transfected into LoVo cells. Western blotting analysis was used to screen for the group which had the highest inhibitory rate; the cells were subjected to wound closure assay and transwell assay to determine the migration of LoVo cells after stable transfection. Results： Western blotting analysis showed that ceils transfected with shRNA/324 vector had the strongest inhibition of PI3K p85α protein,with the inhibition rate being 77%, and cells in this group were used for wound closure assay and transwell assay. Wound closure assay showed that the healing ability of PI3K p85α-depleted cells was significantly lower than that of the control cells （P〈0. 05）. Transwell assay showed that the cells passing the polycarbonate membranes was significantly less in PI3K p85α-knoekdown cells than in the control cells （P〈0.05）. Conclusion：Depletion of PI3K p85a protein can obviously inhibit the migration of LoVo cells. PIaK p85α may be a new therapeutic target /or treatment of eolorectal cancer metastasis.