p19细胞向心肌样细胞分化过程中NRD1基因表达的变化

Changes of NRD1 gene expression during the differentiation of p19 cells to cardiac myocytes

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4d，续用生长培养基贴壁培养至16d，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、Western blot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 changes of nardilysm, N-arginine dibasic convertase （NRD1）, gene mRNA and protein expression during the differentiation of p19 cells to eardiae myocytes. Methods p19 Cells were cultured with DMSO in suspension for 4 days to form cell aggregation. Then the aggregates were cultured on gelatin-coated culture dishes without DMSO up to 16 days. The beating of the cells was observed. Western blot of cardiac troponin I（cTnI） were used to identify the cells differentiating into cardiac-like cells. Total RNA and protein were extraeted from p19 cells during the process of differentiation at various time points. The levels of NRD1 mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results After induced by DMSO for 4 days in suspension,spontaneous and rhythmically beating cells were seen on the 8th day, which were cTnI-positive. When p19 cells were induced to differentiate into cardiac myocytes, both the expression level of NRD1 gene mRNA and protein were up-regulated gradually during 0-10 clays of differentiation. The NRD1 gene mRNA and protein expression were obviously up-regulated. There were significant differences among the different days （P〈0.05）. From the 12th to 16th day, both the mRNA and protein expression of NRD1 gene were kept in a higher level, and there was no significant difference among different days （P〉0.05）. Conclusion Both the expression of NRD1 gene and protein were up-regulated in the process of the differentiation of p19 cells into cardiac myocytes, which might be involved in the differentiation of cardiac rnyocytes.