CircRNA_100395 Inhibits Expression of Myocardial Fibrosis-related Genes in Human Atrial Myofibroblasts via Sponging miR-144-3p
- VernacularTitle:CircRNA_100395 通过结合miR-144-3p 抑制心肌成纤维细胞中纤维化相关基因的表达
- Author:
Yi-hong WEN
1
;
Zhen-zhen YANG
1
;
Ming ZHANG
2
;
Jie-ning ZHU
2
;
Ying YANG
3
;
Yong-heng FU
2
;
Xian-hong FANG
2
;
Zhi-xin SHAN
4
Author Information
1. School of Medicine, South China University of Technology, Guangzhou 510006, China
2. Guangdong Provincial key Laboratory of Clinical Pharmacology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China
3. School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China
4. School of Medicine, South China University of Technology, Guangzhou 510006, China; School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China; Guangdong Provincial key Laboratory of Clinical Pharmacology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China; The Second school of Clinical Medicine, Southern Medical University, Guangzhou 510280, China
- Publication Type:Journal Article
- Keywords:
myocardial fibrosis;
human atrial myofibroblast;
circular RNA;
microRNA
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2020;41(6):875-883
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To investigate the effect of circRNA_100395 on myocardial fibrosis and explore its mechanism. 【Methods】 Circular RNA(circRNA) microarray was performed to show the differential expression of circRNAs in the left atrial appendage between patients with long-standing persistent atrial fibrillation(AF) and the healthy controls, and the expression of circRNA_100395 was detected by RT-qPCR, as well as its distribution in the cytoplasm and nucleus of human atrial fibroblasts(HAFs). Expression of circRNA_100395 and its host gene KLHL20 was detected in angiotensin Ⅱ(Ang-Ⅱ) -induced HAFs by RT-qPCR assay. Actinomycin D treatment was performed to test the RNA stability of circRNA_100395 in HAFs. Over-expression of circRNA_100395 was achieved in HAFs with infection of the recombinant circRNA_100395 adenovirus(rAd-circRNA_100395). The expression of Col1a1, Col3a1 and Acta2 was detected in HAFs by RT-qPCR and Western blot assay, respectively. Dual luciferase reporter assay and RNA pull-down assay were performed to identify the interaction between circRNA_100395 and miR-144-3p. 【Results】 Masson staining revealed that myocardial fibrosis was markedly increased in the left atrial appendage of AF patients(P < 0.01). CircRNA_100395 was decreased in the left atrial appendage of AF patients(P < 0.05), but was increased in Ang-II-induced HAFs(P < 0.05), with high enrichment in the cytoplasm of HAFs. CircRNA_100395 was more stable than mRNA of its host gene of KLHL20 when it was subjected to actinomycin D treatment. Overexpression of circRNA_100395 significantly inhibited the expression of fibrosis-related genes in HAFs. Bioinformatics predicted the potential binding site of miR-144-3p in circRNA_100395, and the specific interaction between circRNA_100395 and miR-144-3p was confirmed by the dual luciferase reporter assay and RNA pull-down assay, respectively. Overexpression of miR-144-3p contributed to the expression of fibrosis-related genes in HAFs, while circRNA_100395 could abolish the pro-fibrosis effect of miR-144-3p. 【Conclusion】 CircRNA_100395 exhibits an anti-fibrosis effect in HAFs through sponging miR-144-3p.
