Nkx2-5 gene promotes expression of cardiac markers in 5-azacytidine-induced monolayer P19 cells / 第二军医大学学报

ABSTRACT

Objective: To study the role of Nkx2-5 gene in 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells. Methods: The experiment was divided into two groups an experimental group and a control group. The cells in the experimental group were P19 cells stably expressing Nkx2-5 gene, and cells in the control group were P19 cells. Under the monolayer-culture condition, the cells of two groups were induced by 5-azacytidine (1 μmol/L). The growth of cells were observed by inverted microscope. On the 4th day, 8th day, 12th day and 16th day after induction, RT-PCR was used to detect the expression of GATA-4, α-MHC and ANP gene. Results: In control group, there was no ANP expression after induction; GATA-4 expression was seen on the 8 th day, 12th day, and 16th day after induction; and α-MHC expression was found on the 12th day and 16 th day. In experimental group, the expression of GATA-4 was detected on the 4th day, 8th day, 12th day and 16 th day after induction; Alpha-MHC and ANP expression was noticed on the 8th day, 12th day and 16th day after induction. RT-PCR results showed that the expression of GATA-4 and α-MHC in the experimental group was earlier than that in the control group. And at all time points of observation, the expression of GATA-4 and α-MHC in the experimental group was significantly increased compared with that in the control group (P<0. 05,P<0. 01), except for α-MHC expression on the 12 th day. Conclusion: Nkx2-5 gene can promote 5-azacytidine-induced differentiation of monolayer P19 cells into myocardial cells.