Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Mar 14, 2019; 25(10): 1210-1223
Published online Mar 14, 2019. doi: 10.3748/wjg.v25.i10.1210
Short hairpin RNA-mediated knockdown of nuclear factor erythroid 2-like 3 exhibits tumor-suppressing effects in hepatocellular carcinoma cells
Miao-Mei Yu, Yue-Hua Feng, Lu Zheng, Jun Zhang, Guang-Hua Luo
Miao-Mei Yu, Yue-Hua Feng, Lu Zheng, Jun Zhang, Guang-Hua Luo, Comprehensive Laboratory, the Third Affiliated Hospital of Soochow University, Changzhou 213003, Jiangsu Province, China
Author contributions: Yu MM and Luo GH designed this study; Feng YH and Zhang J performed the experiments; Yu MM, Feng YH, and Zheng L wrote the manuscript; Yu MM and Luo GH performed the bioinformatic prediction; Yu MM and Luo GH conducted statistical analysis; all authors read and approved the final manuscript.
Supported by the Changzhou High-Level Medical Talents Training Project, No. 2016ZCLJ002.
Institutional review board statement: The clinicopathological features involved in this study are all from the TCGA database, thus, no institutional review board document is provided.
Conflict-of-interest statement: None declared.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines and prepared the manuscript accordingly.
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/
Corresponding author: Guang-Hua Luo, MD, PhD, Professor, Senior Researcher, Comprehensive Laboratory, the Third Affiliated Hospital of Soochow University, No. 185, Juqian Street, Tianning District, Changzhou 213003, Jiangsu Province, China. shineroar@163.com
Telephone: +86-519-68870619 Fax: +86-519-86621235
Received: January 4, 2019
Peer-review started: January 4, 2019
First decision: January 23, 2019
Revised: February 13, 2019
Accepted: February 15, 2019
Article in press: February 16, 2019
Published online: March 14, 2019
Abstract
BACKGROUND

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with high mortality-to-incidence ratios. Nuclear factor erythroid 2-like 3 (NFE2L3), also known as NRF3, is a member of the cap ‘n’ collar basic-region leucine zipper family of transcription factors. NFE2L3 is involved in the regulation of various biological processes, whereas its role in HCC has not been elucidated.

AIM

To explore the expression and biological function of NFE2L3 in HCC.

METHODS

We analyzed the expression of NFE2L3 in HCC tissues and its correlation with clinicopathological parameters based on The Cancer Genome Atlas (TCGA) data portal. Short hairpin RNA (shRNA) interference technology was utilized to knock down NFE2L3 in vitro. Cell apoptosis, clone formation, proliferation, migration, and invasion assays were used to identify the biological effects of NFE2L3 in BEL-7404 and SMMC-7721 cells. The expression of epithelial-mesenchymal transition (EMT) markers was examined by Western blot analysis.

RESULTS

TCGA analysis showed that NFE2L3 expression was significantly positively correlated with tumor grade, T stage, and pathologic stage. The qPCR and Western blot results showed that both the mRNA and protein levels of NFE2L3 were significantly decreased after shRNA-mediated knockdown in BEL-7404 and SMMC-7721 cells. The shRNA-mediated knockdown of NFE2L3 could induce apoptosis and inhibit the clone formation and cell proliferation of SMMC-7721 and BEL-7404 cells. NFE2L3 knockdown also significantly suppressed the migration, invasion, and EMT of the two cell lines.

CONCLUSION

Our study showed that shRNA-mediated knockdown of NFE2L3 exhibited tumor-suppressing effects in HCC cells.

Keywords: Nuclear factor erythroid 2-like 3, Hepatocellular carcinoma, The Cancer Genome Atlas, Short hairpin RNA, Epithelial-mesenchymal transition

Core tip: Nuclear factor erythroid 2-like 3 (NFE2L3), as a key regulator of the cellular stress response, is involved in the regulation of various tumors, whereas its role in hepatocellular carcinoma (HCC) has not been elucidated. Our present study identified that NFE2L3 was closely associated with the grade and stage of HCC patients based on The Cancer Genome Atlas database. Furthermore, our study showed that short hairpin RNA-mediated knockdown of NFE2L3 exhibited tumor-suppressing effects in HCC cells.