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DOI: 10.1016/j.aohep.2021.100345
Open Access
Pre-proof, online 1 April 2021
MiR-20b-5p promotes hepatocellular carcinoma cell proliferation, migration and invasion by down-regulating CPEB3
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Zhuokai Li1,#, Wu Lvzhong2,#, Wei Tan1, Kun Zhang1, Qiaomei Lin1, Jinde Zhu1, Tu Chaoyong1, Xinliang Lv1, Chuan Jiang1,
Corresponding author
jiangchuan737@163.com

Corresponding Author: Department of Hepatobiliary and Pancreatic Surgery, Zhejiang University School of Medicine, Lishui Hospital, Lishui Municipal Central Hospital, Fifth Affiliated Hospital of Wenzhou Medical University, No.289 Kuocang Road, Lishui, 323000, Zhejiang Province, China
1 Department of Hepatobiliary and Pancreatic Surgery, Zhejiang University School of Medicine, Lishui Hospital, Lishui Municipal Central Hospital, Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang Province, China
2 Department of General Surgery, Qingyuan County People’s Hospital, Lishui, Zhejiang Province, China
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Abstract

Introduction and Objectives: This study aimed to explore the functional mechanism of the miRNA-20b-5p/cytoplasmic polyadenylation element binding protein 3 (miR-20b-5p/CPEB3) axis in hepatocellular carcinoma (HCC) so as to provide a new idea for targeted therapy of HCC.

Materials and Methods: Bioinformatics analysis was employed to obtain markedly differentially expressed miRNAs and mRNAs in The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset, so as to find target miRNA and its target mRNA. Real-time quantitative PCR was conducted to detect miR-20b-5p and CPEB3 mRNA expression. Western blot was performed to determine CPEB3 protein expression. Dual-luciferase reporter assay was carried out to verify the targeting relationship between miR-20b-5p and CPEB3. Cell counting kit-8 assay, wound healing assay, Transwell invasion assay and flow cytometry were conducted to evaluate the proliferation, migration, invasion and apoptosis of HCC cells.

Results: Bioinformatics analysis suggested that miR-20b-5p and CPEB3 were markedly highly and lowly expressed, respectively, in HCC tissue in TCGA-LIHC dataset. Over-expressing miR-20b-5p facilitated the proliferation, migration and invasion, and suppressed the apoptosis of HCC cells. Dual-luciferase reporter assay validated that there was a targeting relationship between miR-20b-5p and CPEB3. The inhibitory effect of CPEB3 over-expression on HCC cell proliferation, migration and invasion was reversed by over-expressing miR-20b-5p.

Conclusions: The present study proved that miR-20b-5p promotes HCC cell proliferation, migration and invasion by inhibiting CPEB3 expression, which may provide a theoretical basis for the prognosis and treatment of HCC patients.

Keywords:
miR-20b-5p
CPEB3
hepatocellular carcinoma
proliferation
migration
invasion
Abbreviations:
(HCC)
(miRNAs)
(3’-UTR)
(CPEB)
(RRMs)
(EGFR)
(qRT-PCR)
(CCK-8)
(RIPA)
(SDS-PAGE
(WT)
(MUT)
(EDTA)
(SD)
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They have the equal contribution, co-first author

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