Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 7, 2020; 26(5): 478-498
Published online Feb 7, 2020. doi: 10.3748/wjg.v26.i5.478
lncRNACNN3-206 activates intestinal epithelial cell apoptosis and invasion by sponging miR-212, an implication for Crohn's disease
Na Li, Rui-Hua Shi
Na Li, Rui-Hua Shi, Medical School of Southeast University, Nanjing 210009, Jiangsu Province, China
Na Li, Rui-Hua Shi, Department of Gastroenterology, Zhongda Hospital, Affiliated Hospital of Southeast University, Nanjing 210009, Jiangsu Province, China
Na Li, Department of Biomedical Science, Mercer University School of Medicine, Savannah, GA 31404, United States
Author contributions: Shi RH supervised the projects and designed experiments; Li N performed all the experiments; Li N and Shi RH collected normal ileal epithelial tissues and ileal epithelial tissues from patients with Crohn's disease; Li N summed up the experimental results and prepared the first manuscript; and Shi RH made revision.
Supported by Postgraduate Research and Practice Innovation Program of Jiangsu Province, No. KYCX18_0174.
Institutional review board statement: This research was approved by the Ethics Committee of Zhongda Hospital, Affiliated to Southeast University.
Institutional animal care and use committee statement: This research was approved by Animal Experimental Ethical Inspection of Southeast University, NO. 20190227011.
Conflict-of-interest statement: The authors do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (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: Rui-Hua Shi, PhD, Professor, Chief Physician, Department of Gastroenterology, Zhongda Hospital, Affiliated Hospital of Southeast University, Dingjiaqiao Road 87, Nanjing 210009, Jiangsu Province, China. ruihuashi@126.com
Received: October 9, 2019
Peer-review started: October 9, 2019
First decision: December 5, 2019
Revised: December 20, 2019
Accepted: January 8, 2020
Article in press: January 8, 2020
Published online: February 7, 2020
Processing time: 120 Days and 16.1 Hours
Abstract
BACKGROUND

Statistics indicate that the incidence of Crohn’s disease (CD) is rising in many countries. The poor understanding on the pathological mechanism has limited the development of effective therapy against this disease. Previous studies showed that long noncoding RNAs (lncRNAs) could be involved in autoimmune diseases including CD, but the detailed molecular mechanisms remain unclear.

AIM

To identify the differentially expressed lncRNAs in the intestinal mucosa associated with CD, and to characterize their pathogenic role(s) and related mechanisms.

METHODS

The differential expression of lncRNAs was screened by high-throughput RNA sequencing, and the top candidate genes were validated in an expanded cohort by real-time PCR. The regulatory network was predicted by bioinformatic software and competitive endogenous RNA analysis, and was characterized in Caco-2 and HT-29 cell culture using methods of cell transfection, real-time PCR, Western blotting analysis, flow cytometry, and cell migration and invasion assays. Finally, these findings were confirmed in vivo using a CD animal model.

RESULTS

The 3' end of lncRNACNN3-206 and the 3’ UTR of Caspase10 contain high-affinity miR212 binding sites. lncRNACNN3-206 expression was found to be significantly increased in intestinal lesions of CD patients. Activation of the lncRNACNN3-206-miR-212-Caspase10 regulatory network led to increased apoptosis, migration and invasion in intestinal epithelial cells. Knockdown of lncRNACNN3-206 expression alleviated intestinal mucosal inflammation and tissue damage in the CD mouse model.

CONCLUSION

lncRNACNN3-206 may play a key role in CD pathogenesis. lncRNACNN3-206 could be a therapeutic target for CD treatment.

Keywords: Crohn’s disease; Microarray; lncRNACNN3-206; Gene regulation; Cell migration and invasion; miR-212

Core tip: We describe new findings on the overexpression of lncRNACNN3-206 in intestinal lesions of Crohn’s disease (CD) patients by high throughput sequencing. Moreover, forced overexpression of lncRNACNN3-206 led to apoptosis of Caco-2 and HT-29 cells. Since the 3' end of lncRNACNN3-206 and the 3’ UTR of Caspase10 contain high-affinity binding sites of miR-212, whereby lncRNACNN3-206 regulates Caspase10 expression and cell apoptosis/invasion through miR-212 sponging. This mechanism was supported by results from cell lines and the CD mouse model. In conclusion, lncRNACNN3-206 may play a key role in CD pathogenesis, and could be a therapeutic target for CD treatment.