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miR-9-5p 通过靶向 ESR1 促进肝癌细胞的增殖、迁移和侵袭。

miR-9-5p facilitates hepatocellular carcinoma cell proliferation, migration and invasion by targeting ESR1.

机构信息

Department of Hepatobiliary Surgery, Tangshan Gongren Hospital, No. 27 Cultural Road, Lubei District, Tangshan, 063000, Hebei, China.

Shanghai Engineering Research Center of Pharmaceutical Translation, Shanghai, 200231, China.

出版信息

Mol Cell Biochem. 2021 Feb;476(2):575-583. doi: 10.1007/s11010-020-03927-z. Epub 2020 Oct 27.

Abstract

The study aimed to explore the relationship between miR-9-5p and ESR1, and clarify the underlying functional mechanism in the occurrence and development of hepatocellular carcinoma (HCC). Expression data including miRNAs and mRNAs of HCC downloaded from TCGA database were processed for differential analysis, and corresponding clinical data were collected for survival analysis to identify the target miRNA miR-9-5p. Bioinformatics databases were applied for predicting downstream target mRNAs of miR-9-5p. qRT-PCR was used to evaluate expression of miR-9-5p. Western blot was used to detect protein expression of ESR1. MTT, wound healing assay and Transwell assay were used to detect HCC cell proliferation, migration and invasion, respectively. Dual-luciferase reporter gene assay was used to identify the targeting relationship between miR-9-5p and ESR1. Research suggested that miR-9-5p was highly expressed in HCC cells but ESR1 was poorly expressed. Overexpression of miR-9-5p could improve the proliferation, invasion and migration of cells. Dual-luciferase reporter assay showed that ESR1 was the downstream target of miR-9-5p in HCC. Overexpression of miR-9-5p markedly reduced ESR1 mRNA and protein levels in HCC cells, whereas inhibition of miR-9-5p expression produced the contrary results. Silencing ESR1 could noticeably reverse the effect of miR-9-5p knockdown on the proliferation, migration and invasion of HCC cells. As an oncogene, miR-9-5p fostered the proliferation, migration and invasion of HCC cells by targeting and inhibiting ESR1 expression.

摘要

本研究旨在探讨 miR-9-5p 与 ESR1 之间的关系,并阐明其在肝细胞癌(HCC)发生和发展中的潜在功能机制。从 TCGA 数据库中下载 HCC 的 miRNA 和 mRNA 表达数据进行差异分析,并收集相应的临床数据进行生存分析,以确定靶 miRNA miR-9-5p。生物信息学数据库用于预测 miR-9-5p 的下游靶 mRNA。qRT-PCR 用于评估 miR-9-5p 的表达。Western blot 用于检测 ESR1 蛋白的表达。MTT、划痕愈合实验和 Transwell 实验分别用于检测 HCC 细胞的增殖、迁移和侵袭。双荧光素酶报告基因实验用于鉴定 miR-9-5p 与 ESR1 之间的靶向关系。研究表明,miR-9-5p 在 HCC 细胞中高表达,而 ESR1 表达水平较低。miR-9-5p 的过表达可促进细胞的增殖、侵袭和迁移。双荧光素酶报告基因实验显示,ESR1 是 HCC 中 miR-9-5p 的下游靶基因。miR-9-5p 的过表达显著降低了 HCC 细胞中 ESR1 的 mRNA 和蛋白水平,而抑制 miR-9-5p 的表达则产生相反的结果。沉默 ESR1 可明显逆转 miR-9-5p 敲低对 HCC 细胞增殖、迁移和侵袭的影响。作为一种致癌基因,miR-9-5p 通过靶向并抑制 ESR1 的表达,促进 HCC 细胞的增殖、迁移和侵袭。

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