WWTR1对结直肠癌细胞凋亡及侵袭能力的影响

摘要

Objective:To investigate the effect and its mechanism of WWTR1 on the apoptosis and invasion of colorectal cancer cells.Methods:The expression of WWTR1 proteins in human colorectal cancer LOVO, SW480, HCT116 cells were detected by Western blot.NC-siRNA and WWTR1-siRNA were transfected into SW480 cells, without any treatment as control group,the expression of WWTR1 protein after 48 h were detected by Western blot.Cell apoptosis was detected by flow cytometry.BioCoat Matrigel invasion chamber kit was used to detecte the invasion of cells.The expression of apoptosis protein Cleaved caspase3, invasion related protein MMP-2 and MMP-9, Wnt/β-catenin signaling pathway related proteins β-catenin and Cyclin D1 protein were detected by Western blot.Results:The expression of WWTR1 protein in SW480 cells was highest,selected as a follow-up study.RNAi can significantly reduce the expression of WWTR1 protein.Compared with the control group and NC-siRNA group, the apoptosis rate and expression of Cleaved caspase3 protein in WWTR1-siRNA group were increased significantly, the number of cell invasion and MMP-2, MMP-9,β-catenin, Cyclin D1 protein expression were decreased obviously (P ＜0.01).Conclusion:The expression of RNAi silencing WWTR1 may inhibit the apoptosis and decrease the invasiveness of SW480 cells by inhibiting the Wnt/β-catenin signaling pathway.%目的:探讨含有WW结构域的转录调节子1(WWTR1)对结直肠癌细胞凋亡、侵袭能力的影响及机制.方法:Western blot检测人结直肠癌LOVO、SW480、HCT116细胞中WWTR1的蛋白表达;NC-siRNA和WWTR1-siRNA转染SW480细胞,不做任何处理的细胞作为空白对照组,48 h后收集细胞,Western blot检测各组细胞中WWTR1的蛋白表达;流式细胞仪检测细胞凋亡情况;BioCoat Matrigel invasion chamber试剂盒检测细胞的侵袭能力;Western blot检测凋亡相关蛋白Cleaved caspase3、侵袭相关蛋白MMP-2和MMP-9及Wnt/β-catenin信号通路相关蛋白β-catenin和Cyclin D1的表达情况.结果:SW480细胞中WWTR1的蛋白表达最高,选择作为后续的研究对象;RNAi能显著降低WWTR1的蛋白表达;与对照组及NC-siRNA组比较,WWTR1-siRNA组细胞凋亡率及Cleaved caspase3蛋白表达显著升高,细胞侵袭数及MMP-2、MMP-9、β-catenin、Cyclin D1蛋白表达显著降低(P＜0.01).结论:RNAi沉默WWTR1的表达可通过抑制Wnt/β-catenin信号通路诱导SW480细胞凋亡,降低细胞的侵袭能力.