文章摘要
陶元 王瑞敏 唐萄 杨恬 彭惠民 郭海英 星懿展.腺病毒介导的Wnt10b 对HaCaT 细胞迁移的作用及机制[J].,2015,15(3):429-432
腺病毒介导的Wnt10b 对HaCaT 细胞迁移的作用及机制
The Effect and Molecular Mechanismof Adenovirus-Mediated Wnt10b on the Migration of HaCaT Cells
  
DOI:
中文关键词: Wnt10b  经典Wnt信号  HaCaT 细胞  迁移
英文关键词: Wnt10b  Canonical Wnt signaling  HaCaT cells  Migration
基金项目:国家自然科学青年基金项目(31100993);重庆市面上项目(cstc2013jcyjA10021)
作者单位
陶元 王瑞敏 唐萄 杨恬 彭惠民 郭海英 星懿展 第三军医大学学员十五营重庆医科大学基础医学院细胞生物学及遗传学教研室 中铁十七局集团有限公司中心医院重症医学科第三军医大学学员七营 第三军医大学基础部细胞生物学教研室 
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中文摘要:
      目的:探讨Wnt10b 对HaCaT 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HaCaT 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,HaCaT 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,HaCaT 细胞中 Wnt信号关键分子茁-catenin 、细胞粘附分子E-cadherin 表达的改变情况。结果:①Ad-GFP-Wnt10b 感染HaCaT 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;②HaCaT细胞经Ad-GFP-Wnt10b 处理后,细胞创 面愈合速度明显增快;③Wnt10b 处理组细胞茁-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HaCaT 细胞迁移,且该效应涉及经典Wnt/茁-catenin 信号的激活及由E-cadherin 介导的细胞粘附性的减 弱。
英文摘要:
      Objective:To research the effect and the related mechanism of Wnt10b on the migration of HaCaT epidermal cells.Methods:After the HaCaT epidermal cells were treated with different adenovirus ( Ad-GFP or Ad-GFP-Wnt10b), the over-expression levels of Wnt10b protein in the cells were examined by immunofluorescence technique. The migration ability of HaCaT cells was detected by scratch assay at different time points. Western blot was used to measure the expression levels of beta-catenin, which is the key Wnt signaling molecule, and the cell adhesion molecules E-cadherin.Results:① The HaCaT cells were successfully infected by Ad-GFP-Wnt10b. After 48 h, the expression level of GFP in the cells increased. The Wnt10b protein was prominently located in the Wnt10b treatment cells shown by the immunofluorescence staining. ② The scrape wound of HaCaT cells healed more quickly in the Ad-GFP-Wnt10b treatment group. ③In comparison with the control group, the expression of beta-catenin protein markedly increased in the Ad-GFP-Wnt10b treatment group. However, the expression of E-cadherin protein was just the opposite.Conclusion:Wnt10b may promote the migration of HaCaT cells, which involved with the activation of the canonical Wnt/beta-catenin signaling and the decline of the cell adhesion ability mediated by E-cadherin.
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