解整合素金属蛋白酶10通过介导Wnt/β-catenin信号通路促进子宫内膜癌细胞上皮-间质转化

A disintegrin and metalloproteinase 10 promotes epithelial-mesenchymal transition of endometrial cancer cells by mediating the Wnt/β-catenin signaling pathway

目的:探讨解整合素金属蛋白酶10(ADAM10)对子宫内膜癌(EC)细胞上皮-间质转化(EMT)的影响及其作用机制。方法:收集84例EC患者的新鲜癌组织及癌旁组织标本,采用qRT-PCR检测癌组织及其癌旁组织中ADAM10 mRNA表达水平。以人EC细胞株HEC-1B作为研究对象,采用ADAM10过表达慢病毒感染HEC-1B细胞,并联合Wnt/β-catenin信号通路特异性抑制剂XAV939进行干预,再将细胞分为空白对照组(blank)、慢病毒阴性对照组(Lv-NC)、ADAM10过表达慢病毒组(Lv-ADAM10)和Lv-ADAM10+XAV939组。采用qRT-PCR检测感染后各组细胞中ADAM10 mRNA表达水平;划痕和Transwell实验检测各组细胞的迁移和侵袭能力;Western blot法分析各组细胞中ADAM10及EMT相关蛋白和Wnt/β-catenin信号通路相关蛋白表达水平。结果:EC患者癌组织中ADAM10 mRNA表达水平显著高于癌旁组织(P<0.05)。与blank组或Lv-NC组比较,Lv-ADAM10组细胞中ADAM10 mRNA和蛋白表达水平以及N-cadherin、Vimentin、β-catenin、Snail、MMP2和MMP9等蛋白表达水平均显著升高(均P<0.05),而E-cadherin蛋白表达水平明显降低(P<0.05),同时细胞迁移和侵袭能力显著增强(均P<0.05)。与Lv-ADAM10组比较,LvADAM10+XAV939组细胞中N-cadherin、Vimentin、β-catenin、Snail、MMP2和MMP9等蛋白表达水平显著降低(均P<0.05),E-cadherin蛋白表达水平明显升高(P<0.05),且细胞迁移和侵袭能力明显降低(均P<0.05)。结论:ADAM10在EC组织中高表达,其过表达可促进HEC-1B细胞的EMT进程,其作用机制可能与激活Wnt/β-catenin信号通路有关。

Objective:To investigate the effect of a disintegrin and metalloproteinase 10(ADAM10) on epithelial-mesenchymal transition(EMT) of endometrial cancer(EC) cells and its mechanism.Methods:Fresh cancer tissues and adjacent tissues from a total of 84 EC patients were collected, and the mRNA expression levels of ADAM10 in cancer tissues and adjacent tissues were detected by qRT-PCR.HEC-1B of human EC cell line was used as the research object.HEC-1B cells were infected with ADAM10 overexpressed lentivirus and treated with XAV939,a specific inhibitor of Wnt/β-catenin signaling pathway, and the cells were divided into blank control group(blank),lentivirus negative control group(Lv-NC), ADAM10 overexpressed lentivirus group(Lv-ADAM10) and Lv-ADAM10+XAV939 group.qRT-PCR was used to detect the mRNA expression level of ADAM10 in various groups after infection.Wound healing and Transwell experiments were used to detect the migration and invasion abilities of cells in various groups.Western blot was used to analyze the protein expression levels of ADAM10,EMT-related proteins and Wnt/β-catenin signaling pathway related proteins in various groups.Results:The mRNA expression level of ADAM10 in cancer tissues of EC patients was significantly higher than that in adjacent tissues(P<0.05).Compared with blank group or Lv-NC group, the mRNA and protein expression levels of ADAM10,and the protein expression levels of N-cadherin, Vimentin, β-catenin, Snail, MMP2 and MMP9 in Lv-ADAM10 group cells were significantly increased(all P<0.05),while the protein expression level of E-cadherin was significantly decreased(P<0.05),and the migration and invasion ability of cells were significantly increased(all P<0.05).Compared with Lv-ADAM10 group, the protein expression levels of N-cadherin, Vimentin, β-catenin, Snail, MMP2 and MMP9 in Lv-ADAM10+XAV939 group cells were significantly decreased(all P<0.05),while the protein expression level of E-cadherin was significantly increased(P<0.05),and the migration and invasion ability of cells were significantly decreased(all P<0.05).Conclusion:ADAM10 was highly expressed in EC tissues.The overexpression of ADAM10 can promote the EMT process of HEC-1B cells, and its mechanism may be related to the activation of Wnt/β-catenin signaling pathway.