许伟先,何志伟,喻超,田舍,陈世裕,潘耀振,孙诚谊
XU Weixian,HE Zhiwei,YU Chao,TIAN She,CHEN Shiyu,PAN Yaozhen,SUN Chengyi

• 1:贵州医科大学
• 2:贵州医科大学附院肝胆外科
• 3:贵州医科大学肝胆胰脾重点实验室
• 4:贵州省肝胆胰脾疾病研究所

摘要(Abstract)：

目的:探讨miR-125b对胰腺癌细胞(PC)侵袭转移能力的影响及其机制。方法:构建miR-125b过表达载体(miR-125b MIMIC)及空白载体(miR-125b NC)并转染PC细胞系PANC-1细胞作为实验组及阴性对照组,设胰腺正常导管上皮细胞作为BC组;采用划痕实验及Transwell实验检测miR-125b对PC细胞侵袭能力的影响,采用激光共聚焦检测细胞骨架的改变、Western blot检测上皮-间质细胞转化(EMT)相关蛋白钙黏附蛋白E(E-cadherin)及波形蛋白(Vimentin)的变化。结果:Transwell实验及细胞划痕实验结果显示,过表达miR-125b能显著抑制PANC-1细胞的侵袭转移能力,差异有统计学意义(P<0.05);激光共聚焦结果显示,过表达miR-125b后,PANC-1细胞骨架形态紊乱,细胞极性消失;Western blot结果显示,过表达miR-125b后,PANC-1细胞EMT相关蛋白E-cadherin下调、Vimentin上调,细胞侵袭能力增强(P<0.05)。结论:miR-125b能显著抑制PC细胞侵袭转移能力,其机制可能与miR-125b抑制PC细胞EMT转换有关。
Objective: To investigate the effect of miR-125 b expression on the invasion and metastasis of pancreatic cancer cells and its mechanism.Methods: The vector overexpressing miR-125 b(miR-125 b MIMIC) and the blank vector(miR-125 b NC) were constructed,and then transfected into pancreatic cancer cell line PANC-1.The wound healing assay and transwell assay were used to test cell migration and invasion.Confocal microscopy was used to observe the changes of cytoskeleton.Western blot was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related proteins including E-cadherin and vimentin.Results: Transwell assay and wound healing assay showed that overexpression of miR-125 b significantly inhibited the migration and invasion of PANC-1 cells(P<0.05).In addition,miR-125 b overexpression in PANC-1 cells led to cytoskeleton disorder and loss of cell polarity.Western blot analysis showed that miR-125 b overexpression remarkably downregulated E-cadherin,but up-regulated Vimentin(P<0.05).Conclusion: miR-125 b expression can significantly inhibit the migration and invasion of pancreatic cancer cells,and its mechanism may be related to the inhibition of EMT.

关键词(KeyWords)： 胰腺癌;miR-125b;侵袭转移;上皮-间质细胞转化;钙黏附蛋白E;波形蛋白
pancreatic cancer;miR-125b;migration and invasion;epithelial-mesenchymal transition;E-cadherin;vimentin

Abstract:

Keywords:

基金项目(Foundation): 国家国际科技合作专项基金资助(2014DFA31420);; 国家自然科学基金资助项目(81560477);; 贵州省科学技术基金[黔科合J字(2015)2013];; 贵州省第四批人才基地基金资助[黔省专合字(2012)94];; 贵州省科学技术厅-贵阳医学院院士工作站肝胆外科分站[黔科合院士站(2015)4013];; 贵州医科大学肝胆胰脾重点实验室项目资助

作者(Author): 许伟先,何志伟,喻超,田舍,陈世裕,潘耀振,孙诚谊
XU Weixian,HE Zhiwei,YU Chao,TIAN She,CHEN Shiyu,PAN Yaozhen,SUN Chengyi

DOI: 10.19367/j.cnki.1000-2707.2019.08.003

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