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

摘要

Objective To explore the influence of miR-200 c on the biological behavior of laryngeal carcinoma Hep-2 cells and determine whether miR-200 c exerts its biological function through peptidyl-prolyl cis/trans isomerase (PIN1) in laryngeal carcinoma. Methods A qRT-PCR assay for the expression of miR-200 c was performed in laryngeal carcinoma tissues. Hep-2 cells were transfected with miR-200 c 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-200 c and PIN1. Western blotting detected the protein expression level of PIN1. Results The expression of miR-200 c in laryngeal carcinoma was significantly increased. miR-200 c 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-200 c. miR-200 c 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-200 c can inhibit the migration ability of laryngeal carcinoma cells and abnormal centrosome amplification by regulating PIN1.%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.