PTBP1 Mediates the Effect of lncRNA RP11-879F14.2 on Suppression of Fibrosis-related Genes in Cardiac Fibroblasts
- VernacularTitle:PTBP1介导长链非编码RNA RP11-879F14.2发挥抑制心肌纤维化的作用
- Author:
Ying YANG
1
;
Jing GUO
2
;
Yi-hong WEN
2
;
Yu-qing HUANG
1
;
Zhi-yao YI
1
;
Jie-ning ZHU
3
;
Xian-hong FANG
3
;
Zhi-xin SHAN
1
Author Information
1. School of Biology and Biological Engineering， South China University of Technology， Guangzhou，510006， China
2. School of Medicine， South China University of Technology， Guangzhou 510006， China
3. Guangdong Provincial Key Laboratory of Clinical Pharmacology//Guangdong Provincial People’s Hospital//Guangdong Academy of Medical Sciences， Guangzhou 510080， China
- Publication Type:Journal Article
- Keywords:
long non-coding RNA;
lncRNA RP11-879F14.2;
PTBP1;
myocardial fibrosis;
cardiac fibroblasts
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2021;42(1):33-41
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of lncRNA RP11-879F14.2 on fibrotic phenotype of cardiac fibroblasts （CFs） and the mechanism involved. MethodsMasson′s trichrome staining was performed to detect the level of myocardial fibrosis in the myocardium of patients with heart failure （HF） and the healthy controls. LncRNAs micro-array was used to detect the expression of lncRNAs in human myocardium samples. Real-time quantitative PCR （RT-qPCR） was performed to verify RP11-879F14.2 expression in the myocardium of HF patients and the healthy controls. The recombinant RP11-879F14.2 adenovirus （rAd-RP11-879F14.2） was used to infect human atrial myofibroblasts （HAFs）. Expression of Col1a1， Col3a1 and Acta2 was detected by RT-qPCR and Western blot assay， respectively. Distribution of RP11-879F14.2 in the nucleus and cytoplasma of HAFs was determined by RT-qPCR assay. According to the results of bio-informatic prediction， dual-luciferase reporter assay was performed to confirm the interaction between RP11-879F14.2 and polypyrimidine tract binding protein 1 （PTBP1）. Effect of PTBP1 knock-down on fibrosis-related genes expression modulated by RP11-879F14.2 in HAFs was determined. ResultsMasson′s trichrome staining showed that the myocardial fibrosis was significantly increased in the myocardium of HF patients. Consistent with lncRNA microarray results， RP11-879F14.2 was found obviously up-regulated in the myocardium of HF patients. Over-expression of RP11-879F14.2 inhibited mRNA and protein expression of myocardial fibrosis-related genes in HAFs. Results of nucleocytoplasmic separation and RT-qPCR assay showed that RP11-879F14.2 mainly distributed in the nucleus of HAFs. Dual-luciferase reporter assay revealed the interaction between RP11-879F14.2 and PTBP1. Over-expression of PTBP1 enhanced PTBP1 expression in HAFs， but knock-down of PTBP1 could reverse the anti-fibrotic effect of RP11-879F14.2 in HAFs. ConclusionPTBP1 mediates the anti-fibrotic effect of RP11-879F14.2 in HAFs.
