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目的:探讨Wnt10b对Ha Ca T表皮细胞迁移的影响及机制。方法:分别用重组腺病毒Ad-GFP、Ad-GFP-Wnt10b感染Ha Ca T表皮细胞,利用细胞免疫荧光技术检测腺病毒Ad-GFP-Wnt10b感染细胞后的过表达情况;采用细胞划痕实验,于不同时间点观察并记录经Wnt10b处理后,Ha Ca T细胞体外迁移功能的改变;进一步采用Westen blot技术检测不同腺病毒处理后,Ha Ca T细胞中Wnt信号关键分子β-catenin、细胞粘附分子E-cadherin表达的改变情况。结果:1Ad-GFP-Wnt10b感染Ha Ca T细胞48 h后,细胞内GFP表达上调,细胞免疫荧光染色显示Wnt10b处理组高表达Wnt10b蛋白;2Ha Ca T细胞经Ad-GFP-Wnt10b处理后,细胞创面愈合速度明显增快;3Wnt10b处理组细胞β-catenin蛋白表达水平显著高于对照组,而E-cadherin蛋白表达水平显著低于对照组。结论:Wnt10b能促进Ha Ca T细胞迁移,且该效应涉及经典Wnt/β-catenin信号的激活及由E-cadherin介导的细胞粘附性的减弱。
Objective: To investigate the effect of Wnt10b on the migration of HaCa T cells and its mechanism. Methods: The HaCa T cells were infected with Ad-GFP and Ad-GFP-Wnt10b recombinant adenovirus and the expression of Ad-GFP-Wnt10b was detected by cell immunofluorescence staining. At different time points, we observed and recorded the in vitro migration of HaCa T cells after treatment with Wnt10b. The Wntten blot was used to detect the expression of β-catenin, Molecular E-cadherin expression changes. Results: Ad-GFP-Wnt10b transfected HaCa T cells 48h after intracellular GFP expression, immunofluorescence staining Wnt10b Wnt10b protein expression of high expression; 2Ha Ca T cells treated with Ad-GFP-Wnt10b, the cell surface The healing rate was significantly increased. The expression of β-catenin in 3Wnt10b group was significantly higher than that in control group, while the expression of E-cadherin protein was significantly lower than that in control group. CONCLUSION: Wnt10b can promote the migration of HaCa T cells, and this effect involves the activation of classic Wnt / β-catenin signaling and the decrease of cell adhesion mediated by E-cadherin.