Circ_0018478 Inhibits the Fibrotic Phenotype of Cardiac Fibroblasts via Encoding Protein HERC4-193
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2022.0615
- VernacularTitle:Circ_0018478通过编码HERC4-193发挥抑制心肌成纤维细胞纤维化表型的作用
- Author:
Jia-xin FENG
1
;
Ji-shen GUO
2
;
Yu LIANG
3
;
Jia-xue JIANG
1
;
Hui LI
4
;
Jin-dong XU
5
;
Xian-hong FANG
6
;
Zhi-xin SHAN
1
Author Information
1. School of Biology and Biological Engineering， South China University of Technology， Guangzhou 510006， China
2. The Second School of Clinical Medicine， Southern Medical University， Guangzhou 510280， China
3. Department of Cardiology， Guangdong Cardiovascular Institute， Guangzhou 510080， China
4. Department of Clinical Laboratory， Guangdong Provincial People’s Hospital， Guangzhou 510080， China
5. Department of Anesthesiology， Guangdong Provincial People’s Hospital， Guangzhou 510080， China
6. Guangdong Provincial Key Laboratory of Clinical Pharmacology， Guangdong Provincial People’s Hospital， Guangzhou 510080， China
- Publication Type:Journal Article
- Keywords:
cardiac fibrosis;
circ_0018478;
translation;
cardiac fibroblast
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(6):995-1004
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of circ_0018478 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe expression of circ_0018478 and its host gene of HECT and RLD domain containing E3 ubiquitin protein ligase 4 （HERC4） in the myocardium of patients with heart failure （HF） （n=28） and healthy donors （n=18） was analyzed by real-time quantitative polymerase chain reaction （RT-qPCR） assay. The distribution of circ_0018478 was identified by fluorescence in situ hybridization （FISH） assay and RT-qPCR assay based on nucleocytoplasmic RNA in human AC16 cardiomyocytes. Actinomycin D and RNase R exonuclease digestion were used to test the stability of circ_0018478 in AC16 cells. RNA and protein expression of fibrosis-related genes was detected in mouse cardiac fibroblasts （mCFs） with adenovirus-mediated over-expression of circ_0018478. EdU staining and transwell migration assay were performed to detect the effects of circ_0018478 on mCFs proliferation and migration activities. The potential circ_0018478-translated protein in mCFs was identified by mass spectrometry （MS） shot-gun assay. HERC4-193 was inhibited by small interfering RNA （siRNA）， and the effect of HERC4-193 knock-down on the fibrotic phenotype of mCFs with over-expression of circ_0018478 was studied. ResultsThe expression of circ_0018478 was up-regulated in the myocardium of HF patients， with no significant difference in its host gene of HERC4. The results of FISH and RT-qPCR assay showed that circ_0018478 was mainly in the cytoplasm of AC16 cardiomyocytes. The characteristic RNA stability of circ_0018478 was verified by Actinomycin D and RNase R assay， respectively. The enforced expression of circ_0018478 suppressed proliferation and migration of mCFs， and inhibited the expression of fibrosis-related genes in mCFs. The results of MS shot-gun assay and Western blotting showed that circ_0018478 could translate protein HERC4-193. Overexpression of the circ_0018478 and protein HERC4-193 could consistently inhibit the fibrotic phenotype of mCFs. Knock-down of HERC4-193 could attenuate the inhibitory effect of circ_0018478 on fibrosis-related gene expression in mCFs （P<0.05）. ConclusionsCirc_0018478 inhibits the fibrotic phenotype of cardiac fibroblasts via translating HERC4-193 protein.
