miR-552-3p通过抑制DACH1促进肝细胞性肝癌细胞增殖、迁移和侵袭

MiR-552-3p promotes hepatocellular carcinoma cell proliferation,migration and invasion by inhibiting DACH1

目的:探讨miR-552-3p在肝细胞性肝癌(hepatocellular carcinoma,HCC)中的表达及其促进细胞增殖和迁移、侵袭的机制。方法:qRT-PCR检测HCC细胞系及人正常肝细胞系中miR-552-3p的表达情况;下载TCGA数据库中HCC样本的微阵列数据,分析miR-552-3p在HCC癌组织及癌旁组织中是否存在表达差异;miR-552-3p mimics以及miR-552-3p inhibitor转染HCC细胞,荧光素酶报告基因验证miR-552-3p与DACH1是否存在靶向关系,CCK-8及Transwell实验检测miR-552-3p及其与DACH1相互作用对于HCC细胞增殖、迁移、侵袭的影响。结果:TCGA数据库中HCC样本的分析结果表明,miR-552-3p在HCC中高表达,同时qRT-PCR显示miR-552-3p在HCC细胞系中的表达水平显著高于人肝细胞系L02;miR-552-3p的高表达促进HCC细胞的增殖、迁移和侵袭,而抑制miR-552-3p表达后则相反;荧光素酶报告实验证实miR-552-3p靶向作用于DACH1的3′-UTR,上调miR-552-3p可抑制DACH1的表达,而下调miR-552-3p则增加DACH1表达;DACH1的过表达可抑制HCC细胞增殖、迁移和侵袭,但同时过表达miR-552-3p后DACH1对于HCC细胞相关功能的抑制即可解除;miR-552-3p正向调控Wnt/β-catenin信号通路的激活。结论:miR-552-3p靶向作用于DACH1促进HCC细胞增殖、迁移和侵袭,并参与调控Wnt/β-catenin信号。可能为HCC的诊疗策略提供潜在靶点。

Objective:To investigate the expression of miR-552-3p in hepatocellular carcinoma(HCC)and its mechanism of promoting cell proliferation,migration and invasion. Methods:qRT-PCR was used to detect the expression of miR-552-3p in HCC cell lines and normal human liver cell lines. The microarray data of HCC samples were downloaded from TCGA database,and the expressions of miR-552-3p in HCC tumor tissues and its adjacent tissues were analyzed to show whether they are different or not;miR-552-3p mimics and miR-552-3p inhibitor transfected HCC cells,and luciferase assays verified whether miR-552-3p targets DACH1,while CCK8 and Transwell assays detected the effect of miR-552-3p and its interaction with DACH1 on HCC cell proliferation,migration,and invasion. Results:The analysis of HCC samples from the TCGA database indicated that miR-552-3p was highly expressed in HCC,and qRT-PCR showed that the expression level of miR-552-3p in HCC cell line was significantly higher than that in human liver cell line L02;the high expression of miR-552-3p promoted the proliferation,migration and invasion of HCC cells,while the inhibition of miR-552-3p reversed these functions;the luciferase reporter assays verified that miR-552-3p targets the 3′-UTR of DACH1. Up-regulation of miR-552-3p expression inhibited DACH1 whereas down-regulation of miR-552-3p increased DACH1 expression;DACH1 overexpression inhibited HCC cell proliferation,migration and invasion,but with miR-552-3p overexpression the DACH1 inhibitions of HCC cell-related functions were relieved;miR-552-3p positively regulated the activation of Wnt/β-catenin signaling pathway. Conclusion:miR-552-3p targeting DACH1 promotes HCC cell proliferation,migration and invasion,and regulates Wnt/β-catenin signaling,which may provide potential targets for HCC diagnosis and treatment strategies.