科研之友
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摘要
AIM: To investigate the effect of circular RNA (circRNA) -100395 on hypertrophy phenotype of cardiomyocytes and the mechanism involved. METHODS: Expression of circRNA-100395 and its host gene Kelch protein-like family member 20 (KLHL20) was detected by RT-qPCR in the myocardium of healthy organ donors (n=8) and heart failure (HF) patients (n=14) . Neonatal mouse ventricular cardiomyocytes (NMVCs) were infected with GFP adenovirus (rAd-GFP) and recombinant circRNA-100395 adenovirus (rAd-circRNA-100395) for 24 h to explore the effect of circRNA-100395 on the expression of β-myosin heavy chain (β-MHC) , skeletal muscle α-Actin (ACTA1) and atrial natriuretic peptide (ANP) . The effect of circRNA-100395 on the size of NMVCs exposed to angiotensin II (Ang II) treatment was evaluated by Phalloidin-iFluor 647 staining. The binding of microRNA-31-5p (miR-31-5p) with circRNA-100395 was verified by dual-luciferase reporter assay and RNA pull-down assay. RT-qPCR and Western blot were used to detect the effect of miR-31-5p on hypertrophy-related gene expression in NMVCs with over-expression of circRNA-100395. RESULTS: The expression of circRNA-100395 and the mRNA expression of its host gene KLHL20 were significantly increased in the myocardium of HF patients (P<0. 05) . Over-expression of circRNA-100395 significantly inhibited the expression myocardial hypertrophy-related genes, including Myh7 (encoding β-MHC protein) , Acta1 and Anp, in NMVCs, and suppressed Ang II-induced increase in the size of NMVCs (P<0. 01) . The results of dual-luciferase reporter assay and RNA pull-down assay confirmed the interaction between miR-31-5p and circRNA-100395. Over-expression of miR-31-5p attenuated the inhibitory effect of circRNA-100395 on cardiomyocyte hypertrophy. CONCLUSION: circRNA-100395 inhibits hypertrophy-related gene expression through binding to miR-31-5p in NMVCs. ? 2022 The Author(s).
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