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Hsa_circRNA_000166通过调控miR-330-5p/ELK1促进结肠癌细胞增殖、迁移和侵袭。

Hsa_circRNA_000166 Promotes Cell Proliferation, Migration and Invasion by Regulating miR-330-5p/ELK1 in Colon Cancer.

作者信息

Zhao Gang, Dai Gong Jian

机构信息

Anorectal Department, Suqian First Hospital, Suqian 223800, People's Republic of China.

Anorectal Department, Nanjing Traditional Chinese Medicine Hospital, Nanjing 210022, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Jun 12;13:5529-5539. doi: 10.2147/OTT.S243795. eCollection 2020.

Abstract

INTRODUCTION

Circular RNAs (circRNAs), a novel class of non-coding RNAs, which are widely expressed in human cells, have essential roles in the development and progression of cancers. The aim of this study is to figure out the role of circ_000166 in colon cancer (CC) development and the signaling pathway involved.

MATERIALS AND METHODS

HT29 and HCT116 cells were transfected with siRNA of circRNA, miRNA mimics and inhibitors. Cell proliferation, migration and invasion were examined using CCK-8 assay and transwell assay, respectively. Luciferase reporter assay was used to validate the targets of circRNA and miRNA. CC cells were implanted into nude mice subcutaneously to detect tumor growth.

RESULTS

hsa_circRNA_000166 was significantly upregulated in the human CC tissue and in the CC cell lines. Knockdown of hsa_circRNA_000166 reduced cell viability, colony formation, migration and invasion in vitro and decreased tumor size and weight in vivo. Luciferase reporter assay revealed that miR-330-5p was the target of circRNA_000166. miR-330-5p could bind to 3' untranslated region (3'UTR) of ELK1 to downregulate both mRNA and protein expression of ELK1. Dual inhibition of circRNA_000166 and miR-330-5p inhibited the suppression of cell proliferation, migration and invasion induced by si-circRNA_000166.

CONCLUSION

The data of this study demonstrated that the hsa_circRNA_000166 could upregulated the expression of gene by sponging miR-330-5p, which may contribute to a better understanding of the regulatory circRNA/miRNA/mRNA network and CC pathogenesis.

摘要

引言

环状RNA(circRNAs)是一类新型的非编码RNA,在人类细胞中广泛表达,在癌症的发生和发展中起着重要作用。本研究的目的是明确circ_000166在结肠癌(CC)发生中的作用及其涉及的信号通路。

材料与方法

用环状RNA的小干扰RNA、微小RNA模拟物和抑制剂转染HT29和HCT116细胞。分别使用CCK-8法和Transwell法检测细胞增殖、迁移和侵袭。荧光素酶报告基因检测用于验证环状RNA和微小RNA的靶点。将CC细胞皮下植入裸鼠体内以检测肿瘤生长。

结果

hsa_circRNA_000166在人类CC组织和CC细胞系中显著上调。敲低hsa_circRNA_000166可降低体外细胞活力、集落形成、迁移和侵袭能力,并减小体内肿瘤大小和重量。荧光素酶报告基因检测显示miR-330-5p是circRNA_000166的靶点。miR-330-5p可与ELK1的3'非翻译区(3'UTR)结合,下调ELK1的mRNA和蛋白表达。circRNA_000166和miR-330-5p的双重抑制可抑制si-circRNA_000166诱导的细胞增殖、迁移和侵袭的抑制作用。

结论

本研究数据表明,hsa_circRNA_000166可通过海绵吸附miR-330-5p上调基因表达,这可能有助于更好地理解环状RNA/微小RNA/信使RNA调控网络和CC发病机制。

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