沉默CML66基因对肾癌细胞增殖、迁移和侵袭的影响及其机制研究

Effect of CML66 gene silencing on the proliferation,migration and invasion of renal cancer cells and its mechanism

目的探讨应用siRNA(small interfering RNA)沉默慢性粒细胞白血病肿瘤抗原66基因(CML66)对人肾癌786-O细胞生物功能及对LIS1/Dynein信号通路的影响。方法将CML66-siRNA予以应用脂质体转染方法转染入人肾癌786-O细胞中,采用蛋白质印迹法检测转染CML66-siRNA后,检测786-O细胞CML66、LIS1和Dynein蛋白的表达。采用CCK8增殖法检测肾癌细胞的增殖水平;采用Transwell法及Matrigel法检测肾癌细胞的迁移、侵袭能力。结果与对照组比较,转染CML66-siRNA后,肾癌细胞中CML66、LIS1和Dynein蛋白表达下降(P<0.01)。siRNA-CML66组转染2、3、4 d后细胞增殖能力较对照组显著降低(P<0.05)。siRNA-CML66转染组中肾癌细胞侵袭及迁移能力较对照组亦明显减弱(P<0.01)。结论CML66在786-O细胞中高表达,沉默CML66能够降低肾癌细胞的迁移、侵袭及增殖能力,其机制可能与LIS1/Dynein信号通路有关。

Objective To explore the effect of silencing chronic myeloid leukemia tumor antigen 66(CML66)gene with small interfering RNA(siRNA)on the biological functions of human renal carcinoma 786-O cells and the LIS1/Dynein signaling pathway.Methods The liposome transfection method was used to transfect CML66-siRNA targeting CML66 gene into human renal carcinoma 786-O cells.After transfected CML66-siRNA was detected by Western blotting,the expression of CML66,LIS1,and Dynein protein in 786-O cells was detected.The CCK8 method was used to detect cell proliferation ability.Transwell and Matrigel methods were used to measure cell migration and invasion ability.Results Compared with the control group,the expression of CML66,LIS1,and Dynein protein in 786-O cells decreased after transfection of CML66-siRNA,and the difference was statistically significant(P<0.01).The cell proliferation ability of the siRNA-CML66 group was significantly lower than that of the control group after 2,3,and 4 days after transfection(P<0.05).The ability of cell migration and invasion in the siRNA-CML66 transfection group was also significantly weaker than that in the control group(P<0.01).Conclusion CML66 is highly expressed in human renal carcinoma 786-O cells.Down-regulation of CML66 gene can reduce the proliferation,migration,and invasion of human renal carcinoma 786-O cells.The mechanism may be related to the LIS1/Dynein signaling pathway.