CircSLC8A1_005 Inhibits the Fibrotic Phenotype of Cardiac Fibroblasts by Encoding Protein
10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2024.0106
- VernacularTitle:CircSLC8A1_005通过编码蛋白抑制心肌成纤维细胞纤维化表型的作用
- Author:
Yating HU
1
;
Yuan GAO
2
;
Huayan WU
1
;
Yu LIANG
3
;
Hui LI
4
;
Jindong XU
5
;
Yupeng LIU
3
;
Zhixin SHAN
1
Author Information
1. School of Medicine， 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
- Publication Type:Journal Article
- Keywords:
cardiac fibrosis;
circular RNA;
circSLC8A1_005;
translation;
cardiac fibroblast
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2024;45(1):35-44
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of circSLC8A1_005 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe effect of adenovirus-mediated overexpression of circSLC8A1_005 on the expression of fibrosis-related genes， collagen type I alpha 1 chain （Col1a1）， collagen type Ⅲ alpha 1 chain （Col3a1） and smooth muscle actin alpha 2 （Acta2）， in mouse cardiac fibroblasts （mCFs） were detected. The proliferation and migration activities of mCFs were detected by EdU and wound-healing assay， respectively. Dual luciferase reporter gene assay was performed to detect the activity of potential internal ribozyme entry site （IRES） in circSLC8A1_005. CircSLC8A1_005-translated protein， SLC8A1-605aa， and its intracellular distribution was identified by Western blot assay. The effect of SLC8A1-605aa protein on transcription activity of Sod2 gene was detected by the dual luciferase reporter gene assay. RNA binding protein immunoprecipitation （RIP） was utilized to verify the interaction between SLC8A1-605aa and superoxide dismutase 2 （Sod2） mRNA. Actinomycin D treatment was used to detect the effect of SLC8A1-605aa on Sod2 mRNA stability in mCFs. ResultsAn efficient adenovirus-mediated overexpression of circSLC8A1_005 was achieved in mCFs. The enforced expression of circSLC8A1_005 suppressed proliferation and migration of mCFs， and inhibited the expression of fibrosis-related genes in mCFs. The dual luciferase reporter gene assay revealed the activities of 2 IRES in circSLC8A1_005. Results of Western blot assay showed that circSLC8A1_005 could translate protein SLC8A1-605aa with the prospected molecular weight of 70 ku， which is predominantly distributed in the nucleus. Overexpression of the circSLC8A1_005 and SLC8A1-605aa could consistently inhibit the fibrotic phenotype of mCFs. SLC8A1-605aa could up-regulate superoxide dismutase 2 （Sod2） expression， but not at the transcriptional level. RIP assay indicated that SLC8A1-605aa could specifically interact with Sod2 mRNA， and the results of actinomycin D assay showed that SLC8A1-605aa could enhance the stability of Sod2 mRNA in mCFs. ConclusionCircSLC8A1_005 inhibits the fibrotic phenotype of cardiac fibroblasts via translating SLC8A1-605aa protein， and SLC8A1-605aa may be a potential target for the treatment of myocardial fibrosis.
