Wnt/β-catenin信号通路中Wnt基因对肾癌细胞的生物学影响

Biological Effects of Wnt in Wnt/β-catenin Signaling Pathway on Kidney Cancer Cell

目的探讨Wnt基因对肾透明细胞癌Caki-2细胞的生物学影响。方法细胞分3组:Caki-2组(空白对照)、shRNA+Wnt+Caki-2组(慢病毒沉默Caki-2细胞中的Wnt基因)、空载体组(实验对照),CCK-8法观察不同时间各组细胞的增殖情况;透射电镜观察沉默Wnt基因对Caki-2细胞凋亡的影响;Transwell小室方法检测各组细胞侵袭能力的变化;实时PCR观察Wnt/β-catenin信号通路中下游基因的变化和凋亡相关基因的变化。结果 shRNA+Wnt+Caki-2组出现了细胞增殖抑制,其增殖能力与其他2组相比差异有统计学意义(P<0.05)。sh RNA+Wnt+Caki-2组表现出明显的细胞凋亡变化,出现凋亡小体。侵袭实验显示,shRNA+Wnt+Caki-2组迁出的细胞数明显低于其他2组(P<0.05);shRNA+Wnt+Caki-2组中Wnt mRNA、β-catenin mRNA和Bcl-2 mRNA的表达水平均低于其他2组(P<0.05)。结论慢病毒沉默Wnt基因,影响了肾透明细胞癌Caki-2细胞的增殖,促进了细胞的凋亡,抑制了Caki-2细胞的侵袭。

Objective To investigate the biological effects of Wnt gene in kidney cancer Caki-2 cells. Methods The Wnt gene was silenced in kidney cancer Caki-2 cells by lentivirus vector. The cell proliferate ability of cells in each group were assayed by CCK-8 kit at different time points. The apoptosis of Caki-2 cells was observed after silencing Wnt gene by transmission electron microscope. The invasion ability of each group cells was tested using Transwell chambers. The genes expression changes of Wnt/β-catenin signaling pathway and apoptosis related gene were determined by realtime PCR. Results Compared with the other two groups, the cell proliferate ability of the cells after silencing Wnt gene was suppressed, and the difference was statistically significant （P 〈 0.05 ）. Apoptosis increased significantly in shRNA＋Caki-2＋Wnt group cells with silencing of Writ gene, and apoptotic body appeared in these cells. In invasive experiment, the number of emigrated cells in shRNA ＋Caki- 2＋Wnt group were significantly lower than other groups （P 〈 0.05 ）. The expression of Writ mRNA, β-catenin mRNA and Bcl-2 mRNA in shRNA＋Caki-2＋Wnt group cells was lower than other groups （P 〈 0.05 ）. Conclusion Silencing of Wnt gene of kidney cancer Caki-2 cells can affect the proliferation rate of the cells, promote the cell apoptosis, and inhibit the invasion ability, which provide certain theoretical basis for the research and development of new drugs and new therapeutic targets.