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Mechanism of the Regulatory Effect of Overexpression of circMTO1 on Proliferation and Apoptosis of Hepatoma Cells via miR-9-5p/NOX4 Axis

Authors Wang J, Tan Q, Wang W, Yu J

Received 1 December 2019

Accepted for publication 27 April 2020

Published 26 May 2020 Volume 2020:12 Pages 3915—3925

DOI https://doi.org/10.2147/CMAR.S240719

Checked for plagiarism Yes

Review by Single-blind

Peer reviewer comments 2

Editor who approved publication: Dr Beicheng Sun


Jinbao Wang,1,* Qingjuan Tan,2,* Weishan Wang,1 Jie Yu3

1General Surgery, Linyi Central Hospital, Linyi, Shandong Province, People’s Republic of China; 2Neonatal Intensive Care Unit, Linyi Central Hospital, Linyi, Shandong Province, People’s Republic of China; 3Department of Anal-Colorectal, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, Hubei Province, People’s Republic of China

*These authors contributed equally to this work

Correspondence: Jie Yu
Department of Anal-Colorectal, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, Hubei Province, People’s Republic of China
Email yujie19797@163.com

Purpose: To investigate the potential role of the circMTO1/miR-9-5p/NOX4 axis in liver cancer.
Materials and Methods: Human genome-wide circrna microarray V2 was used for analyzing the expression profile of circRNAs in human tissue samples. The TargetScan database was used to predict target genes. Gene overexpression and silencing in hepatoma cell lines were achieved by transfecting the cells with suitable constructs. Quantitative real time PCR and Western blotting were used to analyze gene and protein expression levels. CCK-8 analysis was performed to detect cell proliferation and the transwell assay for analyzing cell migration. Annexin V-FITC/PI staining and immunohistochemistry were respectively used to detect apoptosis and protein expression.
Results: CircMTO1 were down-regulated in the liver cancer tissues and cell lines compared to their respective normal controls. TargetScan database screening and dual luciferase assay revealed that circMTO1 was a molecular sponge of miR-9-5p, and NOX4 was the target gene of miR-9-5p. Overexpression of circMTO1 and NOX4 inhibited proliferation and migration of hepatoma cells, while the overexpression of miR-9-5p had the opposite effects. In contrast, overexpression of circMTO1 and NOX4 promoted apoptosis, while that of miR-9-5p decreased the cell apoptosis rates.
Conclusion: Overexpression of CircMTO1 acts as tumor suppressor in liver cancer by sponging miR-9-5p, which upregulates NOX4.

Keywords: circMTO1, miR-9-5p/NOX4 axis, hepatocellular carcinoma, proliferation, apoptosis

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