上调人滋养层细胞表面抗原-2基因对人前列腺癌细胞迁移、侵袭及上皮-间充质转化的影响

Effects of upregulatioon of trophoblast cell-surface antigens 2 on migration, invasion and epithelial-mesenchymaltransition of human prostate cancer cells

目的 观察过表达人滋养层细胞表面抗原-2（TROP-2）基因对人前列腺癌细胞的迁移和侵袭及上皮-间充质转化的影响.方法 将TROP-2基因克隆到真核表达载体pcDNA3.1,构建TROP-2基因过表达真核表达质粒.PC-3细胞分为3组：空白对照组、空载对照组和TROP-2过表达组.转染PC-3细胞后,采用Western blot法观察TROP-2蛋白表达,采用Transwell方法检测癌细胞迁移和侵袭能力,采用Western blot法检测癌细胞E-钙黏蛋白和N-钙黏蛋白表达.结果 成功构建TROP-2基因真核表达质粒.与空白对照组和空载对照组比较,TROP-2过表达组TROP-2蛋白明显升高.Transwell迁移实验结果显示,空白对照组、空载对照组和TROP-2过表达迁移细胞数分别为（132.6±2.2）、（130.8±1.8）和（189.6±2.6）个.与空白对照组比较,差异有统计学意义（P＜0.01）;Transwell侵袭试验显示,空白对照组、空载对照组和TROP-2过表达迁移细胞数分别为（118.16±1.96）、（117.52±1.85）和（166.38±1.65）个.与空白对照组比较,差异有统计学意义（P＜0.01）.Western blot结果显示,与空白对照组比较,TROP-2过表达细胞组E-钙黏蛋白表达降低,而N-钙黏蛋白表达增高.结论 TROP-2过表达可促进入前列腺癌细胞迁移和侵袭能力,其机制可能与促进上皮-间充质转化有关.

Objective To explore the effects of over-expression of trophoblast cell-surface antigens 2 （TROP-2） gene on the migration and invasion of human prostate cancer cells and the action mechanism.Methods The TROP-2 gene was cloned into the eukaryotic expression vector pcDNA3.1,and TROP-2 over-expression of eukaryotic gene expression plasmid was constructed.All PC-3 cells were divided into three groups：the control group （Con-A）,empty vector control group （Con-B） and over-expression group TROP-2 （pcDNA3.1-TROP-2）.After PC-3 cells in three groups were transfected with phosphate buffer （PBS）,vector pcDNA3.1,or pcDNA3.1-TROP-2,the protein levels of TROP-2 were determined by Western blotting,respectively.The migration and invasion ability was evaluated by Transwell.The E-Cadherin and N-Cadherin protein expression in PC-3 cells was examined by Western blotting.Results TROP-2 gene eukaryotic expression vector was successfully constructed.The Western blotting showed that the expression levels of TROP-2 protein were greatly increased in PC-3 cancer cells transfected with pcDNA3.1-TROP-2.The Transwell migration assay revealed that the number of migrating cells in Con-A,Con-B,and pcDNA3.1-TROP-2 groups was 132.6 ±2.2,130.8 ± 1.8 and 189.6±2.6,respectively （P ＜ 0.01）.The Transwell invasion assay indicated that the number of migrating cells in Con-A,Con-B,and pcDNA3.1-TROP-2 groups was 118.16 ± 1.96,117.52 ± 1.85 and 166.38 ± 1.65,respectively （P ＜ 0.01）.As Compared with the Con-A group,the E-cadherin protein expression was decreased,and the N-cadherin protein expression was increased in PC-3 cancer cells transfected with pcDNA3.1-TROP-2.Conclusion TROP-2 overexpression can promote migration and invasion of human prostate cancer cells probably promoting the epithelial-mesenchymal transition.