Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 14, 2020; 26(22): 3034-3055
Published online Jun 14, 2020. doi: 10.3748/wjg.v26.i22.3034
Hsa_circRNA_102610 upregulation in Crohn’s disease promotes transforming growth factor-β1-induced epithelial-mesenchymal transition via sponging of hsa-miR-130a-3p
Juan Yin, Yu-Lan Ye, Tong Hu, Li-Juan Xu, Li-Ping Zhang, Ru-Ning Ji, Ping Li, Qian Chen, Jian-Yun Zhu, Zhi Pang
Juan Yin, Ping Li, Qian Chen, Jian-Yun Zhu, Zhi Pang, Department of Digestive Disease and Nutrition Research Center, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou 215008, Jiangsu Province, China
Yu-Lan Ye, Tong Hu, Li-Juan Xu, Li-Ping Zhang, Zhi Pang, Department of Gastroenterology, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou 215008, Jiangsu Province, China
Ru-Ning Ji, Department of Biomedical Engineering, the Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou 215008, Jiangsu Province, China
Author contributions: Pang Z and Yin J designed this study; Zhu JY guided experimental methods; Ye YL, Hu T, Xu L and Zhang LP collected the samples; Yin J carried out the experiments; Li P, Qian C and Ji RN helped with data analysis; Yin J wrote the manuscript; Pang Z, Ye YL and Zhu JY revised the manuscript; all authors approved the final version of the article.
Supported by the Suzhou Special Project of Diagnosis and Treatment for Key Clinical Disease, No. LCZX201715; the Natural Science Foundation of Jiangsu Province, No. BK20161232; and the Science and Technology Development Fund of Nanjing Medical University, No. NMUB2018215.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at the Ethics Committee of the Affiliated Suzhou Hospital of Nanjing Medical University.
Institutional animal care and use committee statement: No animal experiments were conducted in this study.
Conflict-of-interest statement: The authors declare there are no conflicts of interest.
Data sharing statement: No additional data are available.
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:
Corresponding author: Zhi Pang, PhD, Chief Doctor, Full Professor, Department of Gastroenterology, The North District of the Affiliated Suzhou Hospital of Nanjing Medical University, 242 Guangji Road, Suzhou 215008, Jiangsu Province, China.
Received: February 11, 2020
Peer-review started: February 11, 2020
First decision: March 26, 2020
Revised: April 10, 2020
Accepted: April 24, 2020
Article in press: April 24, 2020
Published online: June 14, 2020
Research background

CircRNAs are considered valuable diagnostic biomarkers for Crohn's disease (CD). Our previous study demonstrated that hsa_circRNA_102610 was upregulated in CD patients. Furthermore, miRNA response element (MRE) analysis suggested the existence of a potential interaction between hsa_circRNA_102610 and hsa-miR-130a-3p. Current evidence supports the views that epithelial-mesenchymal transition (EMT) plays an important role in CD pathogenesis, and that hsa-miR-130a-3p can inhibit transforming growth factor-β1 (TGF-β1)-induced EMT.

Research motivation

Further investigation is required to explore the mechanism of hsa_circRNA_102610 in the pathogenesis of CD.

Research objectives

This study was designed to investigate whether the upregulation of hsa_circRNA_102610 correlates with the degree of inflammation in Crohn's disease. The CALP level, C-reactive protein (CRP) level and erythrocyte sedimentation rate (ESR) were included in the correlation analysis. Furthermore, the roles that hsa_circRNA_102610 may play in the proliferation and EMT of intestinal epithelial cells were studied in normal-derived colon mucosa cell line 460 (NCM460) and human intestinal epithelial cells (HIECs).

Research methods

The relative expression levels of hsa_circRNA_102610 and hsa-miR-130a-3p in CD patients were detected by quantitative reverse transcription-polymerase chain reaction. The proliferation of HIECs and NCM460 cells was detected by cell counting kit-8, EdU staining and cell cycle assays following overexpression or downregulation of hsa_circRNA_102610. Cell proliferation assays were performed as described above in a rescue experiment with hsa-miR-130a-3p mimics. The interaction of hsa_circRNA_102610 and hsa-miR-130a-3p was verified by fluorescence in situ hybridization and dual luciferase reporter assays. The relative expression levels of CyclinD1, SMAD4, E-cadherin, N-cadherin and Vimentin were detected by western blotting following hsa_circRNA_102610 overexpression, TGF-β1-induced EMT or hsa-miR-130a-3p mimic transfection (in rescue experiments).

Research results

Upregulation of hsa_circRNA_102610 was determined to be positively correlated with elevated fecal calprotectin levels in CD (r = 0.359, P = 0.007) by Pearson correlation analysis. Hsa_circRNA_102610 promoted the proliferation of HIECs and NCM460 cells, while hsa-miR-130a-3p reversed the cell proliferation-promoting effects of hsa-circRNA_102610. Fluorescence in situ hybridization and dual luciferase reporter assays showed that hsa_circRNA_102610 directly bound hsa-miR-130a-3p in NCM460 and 293T cells. An inverse correlation between downregulation of hsa-miR-130a-3p and upregulation of hsa_circRNA_102610 in CD patients was observed (r = -0.290, P = 0.024) by Pearson correlation analysis. Moreover, overexpression of hsa_circRNA_102610 promoted SMAD4 and CyclinD1 protein expression, as validated by western blotting. Furthermore, overexpression of hsa_circRNA_102610 promoted TGF-β1-induced EMT in HIECs and NCM460 cells via targeting of hsa-miR-130a-3p, with increased expression of Vimentin and N-cadherin and decreased expression of E-cadherin.

Research conclusions

Hsa_circRNA_102610 upregulation in CD patients could promote the proliferation and EMT of intestinal epithelial cells via sponging of hsa-miR-130a-3p.

Research perspectives

Hsa_circRNA_102610 may serve as a potential target for CD therapy and novel drug research. Externally delivered hsa-miR-130a-3p could possibly act as a sponge of hsa_circRNA_102610.