CIAPIN1通过抑制ERK1/2途径对膀胱尿路上皮癌增殖和转移的作用研究

Study on Effect of CIAPIN1 on Proliferation and Metastasis of Bladder Urothelial Carcinoma by Inhibiting ERK1/2 Pathway

目的:探究CIAPIN1在膀胱尿路上皮癌中的作用及其可能的机制。方法:以膀胱尿路上皮癌细胞系J82、T24、UM-UC-3和人膀胱上皮永生化细胞SV-HUC-1为研究对象,qRT-PCR检测细胞CIAPIN1 mRNA表达水平;Western blot检测细胞CIAPIN1、p-ERK1/2和ERK1/2蛋白表达水平;5-乙炔基-2'-脱氧尿苷(EdU)标记实验检测细胞增殖;细胞划痕实验检测细胞迁移;Transwell实验检测细胞侵袭。结果:与SV-HUC-1组相比,T24、UM-UC-3和J8两组细胞中CIAPIN1 mRNA和蛋白表达水平均明显升高(P<0.05),其中,T24组细胞变化最为明显,因此选取T24进行后续实验。与NC组相比,si-CIAPIN1组EdU阳性细胞数、细胞迁移距离和侵袭细胞数明显降低(P<0.05),p-ERK1/2/ERK1/2蛋白表达水平明显升高(P<0.05);与si-CIAPIN1组相比,si-CIAPIN1+U0126组EdU阳性细胞数、迁移距离和侵袭细胞数均明显升高(P<0.05),p-ERK1/2/ERK1/2蛋白表达水平明显降低(P<0.05)。结论:CIAPIN1通过抑制ERK1/2途径磷酸化促进膀胱尿路上皮癌细胞的增殖和转移。

Objective:To explore the role and possible mechanism of CIAPIN1 in bladder urothelial carcinoma.Methods:Bladder urothelial cancer cell lines J82,T24,UM-UC-3 and human bladder epithelial immortalized cells SV-HUC-1 were used as the research objects.qRT-PCR was used to detect the expression level of CIAPIN1 mRNA in the cells;Western blot was used to detect the expression levels of CIAPIN1 p-ERK1/2 and ERK1/2 protein in cells;5-ethynyl-2'-deoxyuridine(EdU)labeling test was used to detect cell proliferation;cell scratch test was used to detect cell migration;Transwell test was used to detect cell invasion.Results:Compared with SV-HUC-1 group,the expression levels of CIAPIN1 mRNA and protein in cells of T24,UM-UC-3 and J82 groups were significantly increased(P<0.05).Cells in T24 group had the most obvious changes,so T24 cells were selected for subsequent experiments.Compared with the NC group,the number of EdU-positive cells,cell migration distance,and invasive cells in the si-CIAPIN1 group were significantly reduced(P<0.05),and protein expression level of p-ERK1/2/ERK1/2 was significantly increased(P<0.05);Compared with the si-CIAPIN1 group,the number of EdU positive cells,migration distance,and number of invasive cells in the si-CIAPIN1+U0126 group were significantly increased(P<0.05),and the protein expression level of p-ERK1/2/ERK1/2 was significantly reduced(P<0.05).Conclusion:CIAPIN1 promotes the proliferation and metastasis of bladder urothelial cancer cells by inhibiting ERK1/2 pathway phosphorylation.