CircRNA Circ_0120051 Inhibits the Fibrotic Phenotype of Myocardial Fibroblasts via Targeting miR-144-3p/IDH2 Axis
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20240305.004
- VernacularTitle:环状RNA circ_0120051通过miR-144-3p/IDH2轴抑制心肌成纤维细胞的纤维化表型研究
- Author:
Yu LIANG
1
;
Zhiqin HU
2
;
Yihong WEN
3
;
Huayan WU
3
;
Ya WNAG
3
;
Yupeng LIU
1
;
Zhixin SHAN
4
;
Xianhong FANG
1
Author Information
1. Guangdong Cardiovascular Institute//Guangdong Provincial People’s Hospital//Guangdong Academy of Medical Sciences， Southern Medical University， Guangzhou 510080， China
2. Department of Pharmacy， The Fifth Affiliated Hospital of Guangzhou Medical University， Guangzhou， Guangdong 510799， China
3. School of Medicine， South China University of Technology， Guangzhou 510006， China
4. Guangdong Provincial Key Laboratory of Clinical Pharmacology// Guangdong Provincial People’s Hospital// Guangdong Academy of Medical Sciences， Southern Medical University， Guangzhou 510080， China
- Publication Type:Journal Article
- Keywords:
cardiac fibrosis;
circular RNA;
miRNA;
Circ_0120051;
cardiac fibroblast
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2024;45(2):196-205
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the regulatory effect of circular RNA circ_0120051 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe expression of circ_0120051 and its host gene of solute carrier family 8 member A1（SLC8A1） mRNA in the myocardium of healthy organ donors （n=24） and heart failure （HF） patients （n=21） were assessed by real-time quantitative polymerase chain reaction （RT-qPCR） assay. RNA stability of circ_0120051 was identified by RNase R exonuclease digestion assay. The cytoplasmic and nuclear distribution of circ_0120051 in human cardiomyocyte AC16 was detected by RT-qPCR assay. The expression of fibrosis-related genes in mouse cardiac fibroblasts （mCFs） with adenovirus-mediated overexpression of circ_0120051 was detected by RT-qPCR and Western blot assay， respectively. The effect of overexpression of circ_0120051 on the migration activity of mCFs was evaluated by wound-healing assay. RNA co-immunoprecipitation （RIP） was conducted to detect the interaction between circ_0120051 and miR-144-3p. The binding site of miR-144-3p in the 3'-UTR of isocitrate dehydrogenase 2 （Idh2） mRNA was identified by the dual luciferase reporter gene assay. ResultsCirc_0120051 was significantly up-regulated in the myocardium of HF patients， while the mRNA expression of its host gene SLC8A1 was not changed. Circ_0120051 was mainly located in the cytoplasm of human AC16 cells. Results of RNase R exonuclease digestion revealed that circ_0120051 possesses the characteristic stability of circular RNA compared to the linear SLC8A1 mRNA. Overexpression of circ_0120051 could inhibit the expression of fibrosis-related gene in mCFs and mCFs migration. RIP assay confirmed the specific interaction between circ_0120051 and miR-144-3p. Transfection of miR-144-3p mimic could efficiently promote the expression of fibrosis-related genes in mCFs and reverse the inhibitory effect of circ_0120051 on the fibrotic phenotype of mCFs. Results of the dual luciferase reporter gene assay confirmed the interaction between miR-144-3p and the 3'-UTR of Idh2. Transfection of miR-144-3p transcriptionally inhibited Idh2 expression， and overexpression of circ_0120051 enhanced IDH2 expression in mCFs. MiR-144-3p mimic and Idh2 small interfering RNA （siRNA） could consistently reverse the inhibitory effects of circ_0120051 on fibrosis-related genes expression in mCFs and mCFs migration. ConclusionsCirc_0120051 inhibits the fibrotic phenotype of cardiac fibroblasts via sponging miR-144-3p to enhance the target gene of IDH2 expression.
