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

doi:10.3969/j.issn.2095-4344.2016.45.019

Abstract

Abstract:

BACKGROUND: Previous studies have found that the expression level of miR-2135 in differentiated P19 cells is significantly higher than that in undifferentiated P19 cells. However, the effects of miR-2135 on P19 cell proliferation and differentiation into cardiomyocytes as well as cardiomyogenesis remain unclear.
OBJECTIVE: To explore the effect of miR-2135 on P19 cell proliferation and differentiation into cardiomyocytes.
METHODS: P19 cells were transferred with miR-2135 knockdown plasmid or vector plasmid. After 4 days of culture, cell proliferation was detected by MTT assay. Flow cytometry was used to detect cell cycle at 0, 24, 48 hours of culture. P19 cells in the two groups were induced to differentiate into cardiomyocytes. The expression of miR-2135 in differentiated and undifferentiated P19 cells 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.
RESULTS AND CONCLUSION: Expression level of miR-2135 in miR-2135-downexpressed P19 cells was obviously reduced. Compared with control cells, miR-2135 knockdown significantly upregulated P19 cell proliferation (P < 0.05) as well as induced a cell 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 cells 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 cell proliferation, but promotes the differentiation of P19 cells into cardiomyocytes.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Myocytes, Cardiac, MicroRNAs, Tissue Engineering

CLC Number:

R394.2

Li Hui. Effect of miR-2135 on the P19 cell proliferation and differentiation into cardiomyocytes[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(45): 6814-6820.

Figures/Tables 1

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