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目的观察苯乙肼裂解酶1(NRD1)在p19细胞向心肌样细胞诱导分化过程中表达的变化。方法p19细胞经1%二甲基亚砜(DMSO)悬浮诱导培养4 d,续用生长培养基贴壁培养至16 d,观察细胞跳动情况,用Western blot鉴定心肌细胞肌钙蛋白I(cTnI),并采用RT-PCR、Westernblot检测p19细胞中NRD1基因mRNA和蛋白的表达。结果p19细胞于诱导分化的第8天出现自发性节律跳动,cTnI蛋白呈阳性。NRD1基因在分化的第0~10天表达显著增高,各时间点之间差异有统计学意义(P<0.05);第12~16天该基因表达趋于稳定,各时间点之间差异无统计学意义(P>0.05)。结论NRD1基因在p19细胞向心肌细胞分化过程中表达逐渐上调,可能参与心肌细胞的分化和发育。
Objective To investigate the expression of p-ethylenylhydrazine lyase 1 (NRD1) during the differentiation of p19 cells into cardiomyocyte-like cells. METHODS: The p19 cells were cultured in suspension in 1% dimethylsulfoxide (DMSO) for 4 days and then cultured on adherent medium for 16 days. The cell viability was observed and the cTnI expression was detected by Western blot. , And the expression of NRD1 mRNA and protein in p19 cells was detected by RT-PCR and Western blot. Results The p19 cells displayed spontaneous rhythm and cTnI protein positive on the 8th day after differentiation. The expression of NRD1 gene was significantly increased from day 0 to day 10 of differentiation, with statistical significance at each time point (P <0.05). The expression of NRD1 gene tended to be stable from day 12 to day 16 with no statistical difference Significance (P> 0.05). Conclusion The expression of NRD1 gene is up-regulated during the differentiation of p19 cells into cardiomyocytes, which may be involved in the differentiation and development of cardiomyocytes.