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微小RNA-18a通过抑制Dicer 1表达促进体外肝癌细胞迁移和侵袭

microRNA-18a Promotes Cell Migration and Invasion Through Inhibiting Dicer l Expression in Hepatocellular Carcinoma In Vitro.

作者信息

Zhang Xiufen, Yu Bo, Zhang Fuzheng, Guo Zijian, Li Lihua

机构信息

Institute of Oncology, The Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu 214062, China.

Department of Oncology, People's Hospital of Taixing, Taixing, Jiangsu 225400, China.

出版信息

Chin Med Sci J. 2017 Apr 10;32(1):34-3. doi: 10.24920/j1001-9242.2007.005.

Abstract

Objective To investigate the effects of microRNA-18a (miR-18a) on migration and invasion of hepatocellular carcinoma (HCC) cells, and its possible mechanism associated with Dicer l.Methods HepG2 and HepG2.2.15 cells were transfected with miR-18a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay, and cell migration was detected by transwell migration and wound-healing assays. Moreover, luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition, a functional restoration assay was performed to investigate whether miR-18a promotes HCC cell migration and invasion by directly targeting Dicer 1.Results miR-18a inhibitor can suppress the migration and invasion of HCC cells. Furthermore, suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18a inhibitor, restorating these activities to levels similar to the parental HCC cells. Interestingly, suppression of miR-18a in HCC cells resulted in enhanced expression of Dicer l. In addition, the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18a.Conclusion Our findings suggest that miR-18a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.

摘要

目的 探讨微小RNA-18a(miR-18a)对肝癌(HCC)细胞迁移和侵袭的影响及其与Dicer 1相关的可能机制。方法 使用Lipofectamine将miR-18a抑制剂转染至HepG2和HepG2.2.15细胞。通过Transwell侵袭实验评估细胞侵袭能力,通过Transwell迁移实验和伤口愈合实验检测细胞迁移能力。此外,采用荧光素酶报告基因实验鉴定miR-18a是否调控Dicer表达。进行实时逆转录聚合酶链反应(Real-time RT-PCR)和蛋白质免疫印迹法(western blot)分析Dicer 1的表达。另外,进行功能恢复实验以研究miR-18a是否通过直接靶向Dicer 1促进HCC细胞迁移和侵袭。结果 miR-18a抑制剂可抑制HCC细胞的迁移和侵袭。此外,小干扰RNA抑制Dicer 1表达基本消除了miR-18a抑制剂对细胞迁移和侵袭的抑制作用,使这些活性恢复到与亲本HCC细胞相似的水平。有趣的是,抑制HCC细胞中的miR-18a会导致Dicer 1表达增强。此外,荧光素酶实验结果表明miR-18a对Dicer 1有靶向调控作用。结论 我们的研究结果表明,miR-18a通过抑制Dicer 1表达促进HCC细胞的迁移和侵袭。

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