Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 14, 2015; 21(46): 13030-13041
Published online Dec 14, 2015. doi: 10.3748/wjg.v21.i46.13030
Up-regulation of microRNA-210 inhibits proliferation of hepatocellular carcinoma cells by targeting YES1
Weiqi Tan, Seng-Gee Lim, Theresa MC Tan
Weiqi Tan, Theresa MC Tan, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore S117597, Singapore
Seng-Gee Lim, Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore S117597, Singapore
Author contributions: Tan W performed the experiments; Lim SG contributed to the hepatocellular carcinoma data; Tan W and Tan TMC wrote the paper.
Supported by Biomedical Research Council and Ministry of Education (Tier 1) awarded to Tan TM.
Institutional review board statement: The study was reviewed and approved by the National Health Group Domain Specific Review Board.
Institutional animal care and use committee statement: This is not applicable. No animal was used for this study.
Conflict-of-interest statement: To the best of our knowledge, no conflict of interest exists.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Theresa MC Tan, Associate Professor, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, MD7, 8 Medical Drive, Singapore S117597, Singapore. bchtant@nus.edu.sg
Telephone: +65-65163685 Fax: +65-67791453
Received: May 22, 2015
Peer-review started: May 23, 2015
First decision: July 19, 2015
Revised: September 10, 2015
Accepted: October 17, 2015
Article in press: October 20, 2015
Published online: December 14, 2015
Processing time: 201 Days and 6.7 Hours
Abstract

AIM: To determine the expression of microRNA-210 (miR-210) in hepatocellular carcinoma (HCC) and to examine its role using HCC cells.

METHODS: The expression of miR-210 was determined in 21 pairs of HCC samples and the corresponding surrounding non-tumor tissues. The effects of miR-210 on proliferation and cell cycle progression were examined using HepG2 and HuH7 cells. Over-expression and inhibition of miR-210 was achieved by transfection of the cells with miR-210 mimic or inhibitor. Luciferase reporter constructs were used to identify the miR-210 interacting site on Yes1. Yes1 expression was examined after miR-210 transfection, as well as in the HCC samples.

RESULTS: miR-210 was significantly up-regulated by 3.4 fold (P < 0.01) in the tumor samples. The over-expression of miR-210 significantly reduced cell proliferation compared to the mock-treated cells (68.9% ± 7.4% and 53.6% ± 5.0%, P < 0.05 for the HepG2 and HuH7 cells respectively). Analysis of the HuH7 cells transfected with miR-210 mimic by flow cytometry showed that the cells took a longer time to reach the G2/M phase. The interaction between miR-210 and the 3’UTR of the Yes1 transcript was confirmed using a luciferase reporter assay. Over-expression of miR-210 reduced the expression of Yes1 protein in both HuH7 and HepG2 cells. Tumors with a greater than four-fold increase in the expression of miR-210 showed consistently lower expressions of Yes1 in the tumors. In nocodazole-treated cells with a significant G2/M cell population, Yes1 protein was significantly reduced and pre-inhibition of miR-210 in HuH7 cells was able to prevent the reduction of Yes1 protein expression. Knock-down of Yes1 by siRNA also led to reduced cell proliferation (70.8% ± 7.5%, P < 0.05 in the HuH7 cells).

CONCLUSION: Up-regulation of miR-210 inhibits cell proliferation. Yes1 is a target of miR-210 and affects cell proliferation in HCC.

Keywords: MicroRNA-210; Hepatocellular carcinoma; Proliferation; Yes1

Core tip: In this study, miR-210 is significantly up-regulated in hepatocellular carcinoma (HCC). Over-expression of miR-210 decreased cell proliferation and delayed cell cycle progression of HCC cells. The tyrosine kinase Yes1 is shown to be a target of miR-210 and is down-regulated in HCC. Knock-down of Yes1 by siRNA also significantly reduced cell proliferation. These results increase the understanding of the multiple roles of miR-210 in liver cancer growth and metastasis.