Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 21, 2015; 21(19): 5884-5892
Published online May 21, 2015. doi: 10.3748/wjg.v21.i19.5884
Effect of microRNA-1 on hepatocellular carcinoma tumor endothelial cells
Chao Hu, Shi-Qiang Shen, Zhong-Hui Cui, Zu-Bing Chen, Wei Li
Chao Hu, Shi-Qiang Shen, Zhong-Hui Cui, Zu-Bing Chen, Wei Li, Department of General Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
Author contributions: Hu C performed the majority of experiments, analyzed the data and wrote the paper; Shen SQ critically revised the manuscript, and approved the final version of the manuscript submitted for publication; Chen ZB designed the study; Cui ZH performed the minority of experiments; and Li W contributed analytic tools.
Ethics approval: All experiments involving animals and human subjects were designed and performed in compliance with the relevant laws regarding the humane care and use of subjects.
Institutional animal care and use committee: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Renmin Hospital of Wuhan University (IACUC protocol number: SYXK 2014-0013).
Conflict-of-interest: We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.
Data sharing: 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: Shi-Qiang Shen, MD, Department of General Surgery, Renmin Hospital of Wuhan University, 99 Zhangzhidong road, Wuchang district, Wuhan 430060, Hubei Province, China. swsw2218@hotmail.com
Telephone: +86-27-88041911 Fax: +86-27-88042292
Received: November 30, 2014
Peer-review started: December 1, 2014
First decision: December 26, 2014
Revised: January 24, 2015
Accepted: February 13, 2015
Article in press: February 13, 2015
Published online: May 21, 2015
Processing time: 171 Days and 6.7 Hours
Abstract

AIM: To investigate the effect of microRNA-1 (miR-1) on tumor endothelial cells (TECs) of human hepatocellular carcinoma (HCC).

METHODS: MiR-1 specific short hairpin RNA (shRNA) was synthesized and cloned into a recombinant lentiviral vector. TECs were then infected by the miRNA-1-shRNA recombinant lentivirus. TECs were divided into three groups: a control (CON) group consisting of normal TECs without lentiviral infection, a negative control (NC) group consisting of normal TECs infected with a negative control virus, and a micro-down (MD) group consisting of normal TECs infected with the miR-1-inhibition virus containing the target gene. Silencing of miR-1 expression was quantified via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The proliferation of TECs was detected using MTT (Thiazolyl Blue Tetrazolium Bromide) assay; the observations were continued for 5 d, and the optical density value at 490 nm was detected every day. Apoptosis was detected via flow cytometry using Annexin V-APC single staining. The migration and invasion of TECs were detected using transwell assays.

RESULTS: Lentiviral miR-1 shRNA was successfully transduced into TECs, and specifically silenced the expression of miR-1. The results of qRT-PCR showed that the expression of miR-1 was significantly decreased in the MD group (2-ΔΔCt = 0.57 ± 0.14) compared with the CON group (2-ΔΔCt = 1) and the NC group (2-ΔΔCt = 1.05 ± 0.13) (P < 0.01). The results of MTT assay showed that the cell proliferation was all significantly inhibited in the MD group in the 5 days compared with the CON and NC groups (P < 0.01). The results of flow cytometry showed that the apoptosis was significantly increased in the MD group (6.32% ± 0.33%) compared with the CON group (2.03% ± 0.30%) and the NC group (2.18% ± 0.15%) (P < 0.01). The ability of cell migration was significantly inhibited in the MD group (62.0 ± 5.48) compared with the CON group (99.8 ± 3.11) and the NC group (97.2 ± 3.70) (P < 0.01). The ability of invasion of TECs was also significantly inhibited in the MD group (29.8 ± 2.39) compared with the CON group (44.6 ± 3.36) and the NC group (44.4 ± 5.17) (P < 0.01).

CONCLUSION: MiR-1 might be a potential tumor activator. Inhibiting its expression could decrease proliferation, induce apoptosis, and inhibit the migration and invasion of TECs of human HCC.

Keywords: Tumor endothelial cells; Hepatocellular carcinoma; Short hairpin RNA; MicroRNA-1

Core tip: Our study demonstrated that microRNA-1 (miR-1) might be a potential tumor activator. Inhibition of the expression of miR-1 could decrease the proliferation, induce the apoptosis, and inhibit the migration and invasion of tumor endothelial cells of human hepatocellular carcinoma.