Research On The Mechanism Of BMP4/DPF3 Affect On Cardiomyocyte Differentiation In TOF

Background: TOF had been the most common complex congenital heart disease.It had accounted for 3%-5% in all kinds of congenital heart disease.Some theories had been popular recently like the abnormal transference of neural crest cell,the abnormal EMT and the abnormal differentiation of myocardial cell.Many kinds of congenital heart diseases could be caused by the abnormal differentiation of myocardial cell.But so far few researches had been done about whether TOF had the abnormal differentiation of myocardial cell or not.And also few researches had been reported about genetics of TOF at home and abroad.103 children with TOF had been studied.The amount of GATA-4 and NKX2.5 from myocardial cell in RVOT had been tested.It had helped to evaluate whether TOF was related to the abnormal differentiation of myocardial or not.And then RNA screening about the abnormal differentiation of other myocardial cells in those patients had been done to explore the mechanisms.Methods:(1)Using realtime PCR to test the RNA expression of GATA-4 and NKX2.5 in RVOT from TOF group and control group.(2)Using PCR to test 59 associated genes.Contrasting and analyzing the RNA in RVOT expression differences between TOF group and control group.(3)Using 5’-Aza-deoxycytidine with ATRA to induce C3H10T1/2 cell.Studing the behavioristics influence and differentiation regulation that BMP4 and DPF3 gene had made to C3H10T1/2 cell.Results:(1)The mRNA expression of GATA-4 and NKX2.5 in TOF group was significantly increased compared with that of control group,the cases in TOF group with higher mRNA expressions of GATA-4 and NKX2.5 than the Corresponding maximum expression in control group were 65 and 73,60 patients were both involved meanwhile.(2)59 kinds of genes were analyzed between the above 60 patients and the control group while 40 differed significantly.Then those 40 differentially expressed genes had been done correlation analysis respectively with McGoon Index and degree of differentiation into myocardial cell.It had turned out that the differentially expressed genes which had a close relationship with both two of them were DPF3 b,BMP4,CX43 and TBX1.(3)The overexpression of BMP4 could improve the cell migration and invasion.It could inhibit the cell proliferation and improve C3H10T1/2 cell to differentiate to myocardial cell.And it could regulate DPF3 as well which had made the high expression of DPF3.Conclusion:(1)TOF might be associated to the immature differentiation of myocardial cell in RVOT.(2)In TOF there were abnormal expressions of DPF3 b,BMP4,CX43,and TBX1 genes from myocardial cells in RVOT.It had showed statistically significant correlation with McGoon Index of children patients.(3)BPM4 could regulate the behavioristics in the myocardial differentiation model.BMP4 could improve C3H10T1/2 cell to differentiate to myocardial cell and meanwhile increase the high expression of DPF3.