Effect of miR-25 on the differentiation of P19 cells into cardiomyocytes and its mechanism

doi:10.3969/j.issn.2095-4344.2016.11.010

Abstract

Abstract:

BACKGROUND: Previous studies have found that the expression level of miR-25 in differentiated P19 cells is significantly lower than that in undifferentiated P19 cells. 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 cells into cardiomyocytes.
METHODS: P19 cells were cultured and differentiated into cardiomyocytes in vitro. The expression of miR-25 in differentiated and undifferentiated P19 cells was detected by real-time PCR. miR-25-overexpressing P19 cells were constructed by lipofection transfection, and were used to investigate the effect of miR-25 on the differentiation of P19 cells 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 cells with Pax3 shRNAs to silence the expression of Pax3, and investigated the effect of Pax3 on the differentiation of P19 cells into cardiomyocytes.
RESULTS AND CONCLUSION: Expression level of miR-25 in differentiated P19 cells was obviously down-regulated compared with that in undifferentiated P19 cells. miR-25 overexpression promoted the differentiation of P19 cells 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 cells into cardiomyocytes. Taken together, miR-25 promotes the differentiation of P19 cells into cardiomyocytes by targeting Pax3. These findings offer new clues and theoretical basis for cardiomyogenesis and prevention and cure of congenital heart disease.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Li Lian-chong. Effect of miR-25 on the differentiation of P19 cells into cardiomyocytes and its mechanism[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(11): 1584-1590.

Figures/Tables 4

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