Effect of miR-25 on the differentiation of P19 cells into cardiomyocytes and its mechanism / 中国组织工程研究

ABSTRACT

BACKGROUND:Previous studies have found that the expression level of miR-25 in differentiated P19 cels is significantly lower than that in undifferentiated P19 cels. However, the effect of miR-25 on cardiomyogenesis and the relevant mechanism remain unclear. OBJECTIVE: To explore the effect and mechanism of miR-25 on the differentiation of P19 cels into cardiomyocytes. METHODS:P19 cels were cultured and differentiated into cardiomyocytesin vitro. The expression of miR-25 in differentiated and undifferentiated P19 cels was detected by real-time PCR. miR-25-overexpressing P19 cels were constructed by lipofection transfection, and were used to investigate the effect of miR-25 on the differentiation of P19 cels into cardiomyocytes. MicroRNA target analysis tools were used to explore potential targets of miR-25, and dual luciferase reporter assay was used to identify whether the 3’UTR of Pax3 mRNA was a binding target of miR-25. In addition, we transfected P19 cels with Pax3 shRNAs to silence the expression of Pax3, and investigated the effect of Pax3 on the differentiation of P19 cels into cardiomyocytes. RESULTS AND CONCLUSION: Expression level of miR-25 in differentiated P19 cels was obviously down-regulated compared with that in undifferentiated P19 cels. miR-25 overexpression promoted the differentiation of P19 cels into cardiomyocytes. By target prediction analysis, we confirmed that Pax3 was a potential target gene of miR-25. Luciferase assay further confirmed that miR-25 targeted Pax3 directly. Moreover, knockdown of Pax3 promoted the differentiation of P19 cels into cardiomyocytes. Taken together, miR-25 promotes the differentiation of P19 cels into cardiomyocytes by targeting Pax3. These findings offer new clues and theoretical basis for cardiomyogenesis and prevention and cure of congenital heart disease.