miR-200c调控肽基脯氨酰顺反异构酶对喉癌Hep-2细胞生物学行为的影响  

Effect of miR-200c Regulation of Peptidyl-Prolyl Cis/Trans Isomerase on the Biological Behavior of Hep-2 Cells

作　　者：温行 李章富 王辉 孙绍华 郭星[2] 李福才 WEN Xing;LI Zhangfu;WANG Hui;SUN Shaohua;GUO Xing;LI Fucai(Department of Medical Genetics,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China;Department of Otolaryngology,The First Hospital,China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学基础医学院医学遗传学教研室,沈阳110122 [2]中国医科大学附属第一医院耳鼻喉科,沈阳110001

出　　处：《中国医科大学学报》2019年第1期17-22,28,共7页Journal of China Medical University

基　　金：国家自然科学基金(81272969)

摘　　要：目的探究miR-200c对喉癌Hep-2细胞生物学行为的影响以及miR-200c是否通过肽基脯氨酰顺反异构酶(PIN1)在喉癌中发挥其生物学功能。方法实时PCR检测喉癌组织中miR-200c的表达水平;miR-200c相关小RNAs瞬时转染喉癌Hep-2细胞,Transwell实验检测细胞的迁移能力,免疫荧光实验检测细胞中心体的异常扩增;双荧光素酶报告基因系统检测miR-200c与PIN1的结合;Western blotting检测PIN1蛋白的表达水平。结果 miR-200c在喉癌组织中的表达水平显著增高;Transwell结果显示,miR-200c能够抑制Hep-2细胞的迁移,免疫荧光实验显示,miR-200c能够减弱细胞中心体的异常扩增;双荧光素酶报告基因结果证实,PIN1是miR-200c的靶基因之一;Western blotting结果显示,miR-200c可在翻译水平抑制PIN1的表达;miR-200c能够通过调控PIN1PIN1抑制喉癌细胞的迁移能力和减弱细胞中心体的异常扩增。结论 miR-200c通过调控PIN1抑制喉癌细胞的迁移能力,并减弱中心体异常扩增。Objective To explore the influence of miR-200c on the biological behavior of laryngeal carcinoma Hep-2 cells and determine whether miR-200c exerts its biological function through peptidyl-prolyl cis/trans isomerase( PIN1) in laryngeal carcinoma. Methods A qRT-PCR assay for the expression of miR-200c was performed in laryngeal carcinoma tissues. Hep-2 cells were transfected with miR- 200c related small RNAs. Transwell assay detected the migration ability of the cells. Immunofluorescence assay was used to detect the abnormal amplification of the centrosome. A dual luciferase reporter gene system was used to detect the binding ability between miR-200c and PIN1 . Western blotting detected the protein expression level of PIN1. Results The expression of miR-200c in laryngeal carcinoma was significantly increased. miR-200c inhibited the migration of Hep-2 cells and could weaken the abnormal amplification of centrosome. PIN1 was confirmed as one of the target genes of miR-200c. miR-200c inhibited the expression of PIN1 at the translation level and could inhibit Hep-2 cell migration and abnormal centrosome amplification by regulating PIN1 . Conclusion miR-200c can inhibit the migration ability of laryngeal carcinoma cells and abnormal centrosome amplification by regulating PIN1 .

关 键 词：喉癌 MIR-200C 肽基脯氨酰顺反异构酶 迁移 中心体 

分 类 号：R394[医药卫生—医学遗传学]