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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 effect of siRNA targeting PI3Kp85α on the invasion and metastasis of colorectal cancer cells. Methods: Four shRNA interference vectors and one negative control vector were designed and stably transfected into LoVo cells. A group of cells with the highest inhibitory efficiency were screened by Western blot for cell scratch assay and Transwell assay. Results: The expression of PI3Kp85α in LoVo cells transfected with shRNA / 324 interference vector was most significantly decreased with a 77% inhibition rate. The cells were selected as the effective interference group for further experiments. Cell scratch test showed that the scratch healing ability of cells in the interference group was significantly lower than that in the control group (P <0.05). Transwell experiments showed that the number of transmembrane cells in the interference group was significantly less than that in the control group (P <0.05). Conclusion: The loss of PI3Kp85α protein expression can significantly reduce the migration of colorectal cancer LoVo cells, PI3Kp85α may be a new target for the treatment of colorectal cancer metastasis.