TBX1基因可能通过调控Tgfb2基因的表达参与心脏发育及先天性心脏病的发生

TBX1 gene may be involved in heart development and development of congenital heart disease through regulating Tgfb2 gene expression

目的通过检测TBX1基因干扰后转化生长因子β2基因(Tgfb2)表达的变化探讨TBX1基因参与心脏发育及先天性心脏病发生的可能途径。方法根据处理因素不同,将细胞分为NC-siRNA组和TBX1-siRNA组。在大鼠胚胎心肌细胞H9C2中转染TBX1-siRNA,采用实时定量PCR法(qRT-PCR)检测干扰效率;收集转染TBX1-siRNA 48h后的H9C2细胞系,提取总RNA,qRT-PCR法检测干扰后Tgfb2基因mRNA的表达水平。结果(1)qRT-PCR检测心肌细胞TBX1 mRNA的表达:H9C2细胞转染TBX1-siRNA 48h后为0.22±0.02,与NC-siRNA(0.99±0.02)比较,TBX1mRNA表达显著下调,差异有统计学意义(P<0.05)。(2)qRT-PCR检测心肌细胞Tgfb2mRNA的表达:H9C2细胞转染TBX1-siRNA48h后为2.19±0.01,与NC-siRNA(1.00±0.01)比较,Tgfb2mRNA表达明显上调,差异有统计学意义(P<0.05)。结论 TBX1基因表达下调可能通过调控Tgfb2基因的表达参与心脏发育及先天性心脏病的发生。

Objective By detecting the expression of transforming growth factor beta2 gene(Tgfb2)after TBX1 gene interference,we explored the possible way of TBX1 gene involving in cardiac development and development of congenital heart disease.Methods According to the different treatment factors,the cells were divided into NC-siRNA group and TBX1-siRNA group.TBX1-siRNA was transfected in rat embryonic cardiomyocyte H9C2,and the interference efficiency was detected by real time quantitative PCR(qRT-PCR).The H9C2 cell line at 48h after transfection with TBX1-siRNA for was collected to extract the total RNA,and qRT-PCR method was used to detect the expression level of Tgfb2 gene mRNA after interference.Results(1)qRT-PCR was used to detect the expression of TBX1-mRNA in myocardial cells.The expression was(0.22±0.02)at 48h after transfection with TBX1-siRNA.Compared with NC-siRNA(0.99±0.02),the expression of TBX1-mRNA was significantly down regulated,and the difference was statistically significant(P<0.05).(2)The expression of Tgfb2-mRNA in myocardial cells was detected by qRT-PCR,and the expression of H9C2 cells was transfected with TBX1-siRNA.After 48h,the expression of Tgfb2-mRNA was significantly up regulated(2.19±0.01)compared with that of NC-siRNA(1.00±0.01).The difference was statistically significant(P<0.05).ConclusionThe down regulation of TBX1 gene expression may be involved in the development of heart and the development of congenital heart disease by regulating the expression of Tgfb2 gene.