Effect of miR-2135 on the P19 cell proliferation and differentiation into cardiomyocytes

Li Hui

doi:10.3969/j.issn.2095-4344.2016.45.019

Abstract：BACKGROUND:Previous studies have found that the expression level of miR-2135 in differentiated P19 cel s is significantly higher than that in undifferentiated P19 cel s. However, the effects of miR-2135 on P19 cel proliferation and differentiation into cardiomyocytes as wel as cardiomyogenesis remain unclear. <br> OBJECTIVE:To explore the effect of miR-2135 on P19 cel proliferation and differentiation into cardiomyocytes. <br> METHODS:P19 cel s were transferred with miR-2135 knockdown plasmid or vector plasmid. After 4 days of culture, cel proliferation was detected by MTT assay. Flow cytometry was used to detect cel cycle at 0, 24, 48 hours of culture. P19 cel s in the two groups were induced to differentiate into cardiomyocytes. The expression of miR-2135 in differentiated and undifferentiated P19 cel s was detected by quantitative real-time PCR at 0 and 10 days of induction. Levels of cardiac troponin T, atrial natriuretic peptide and myocardial transcription factor were detected using western blot assay at 10 days of induction. <br> RESULTS AND CONCLUSION:Expression level of miR-2135 in miR-2135-downexpressed P19 cel s was obviously reduced. Compared with control cel s, miR-2135 knockdown significantly upregulated P19 cel proliferation (P<0.05) as wel as induced a cel cycle rest in S stage after 24 and 48 hours of culture (P<0.05). PCR findings showed that in the miR-2135 knockdown group, the expression level of miR-2135 at 10 days of induction was significantly higher than that at 0 day of induction (P<0.01). Western blot assay showed that miR-2135-downexpressed P19 cel s had obvious decreases in the protein levels of cardiac troponin T, atrial natriuretic peptide and myocardial transcription factor at 10 days of induction. Our findings confirm that miR-2135 inhibits P19 cel proliferation, but promotes the differentiation of P19 cel s into cardiomyocytes.