论文部分内容阅读
目的利用基因芯片技术观察室间隔缺损(VSD)胎儿心肌组织基因谱表达的变化,对其可能的分子机制进行初步分析。方法病例组为孕中期VSD胎儿,对照组为同胎龄无心脏畸形的难免流产的胎儿,取胎儿心室心肌组织,提取其总RNA,采用安捷伦4×44k人全基因组表达谱芯片观察其心肌组织基因表达谱的变化,对基因芯片数据进行处理和生物信息学分析,差异基因信号通路分析,并用实时PCR方法验证芯片结果。结果芯片筛选发现VSD胎儿心肌组织与正常胎儿心肌组织差异表达基因1 490个,表达差异2倍、3倍、4倍以上的基因数分别为1 314个、157个、19个;信号通路分析差异基因得到18个具有统计学意义的信号通路,其中包括与心脏发育密切相关的信号通路,如:Notch、PI3K/AKT、MAPK信号通路;随机挑选表达差异的5个基因进行验证,结果表明定量PCR检查结果与芯片筛选结果基本相符。结论 VSD胎儿心肌组织与正常胎儿心肌组织差异表达基因与心脏发育密切相关的信号通路(Notch、PI3K/AKT、MAPK信号通路)有关,这为先天性心脏病发生机制的研究奠定了良好的基础。
OBJECTIVE: To observe the change of gene expression in ventricular myocardium with ventricular septal defect (VSD) by gene chip technique and to analyze its possible molecular mechanism. Methods The fetuses in the second trimester VSD fetuses and the control group were unavoidable miscarriage fetuses with the same gestational age without cardiac malformations. The total RNA was extracted from the fetal heart tissues and the myocardial tissues were observed using Agilent 4 × 44k human genome-wide expression microarray Gene expression profile changes, gene chip data processing and bioinformatics analysis, differential gene signal pathway analysis, and real-time PCR method to verify the chip results. Results There were 1 490 differentially expressed genes in VSD fetal heart tissue and normal fetal heart tissue by chip screening. The numbers of genes with 2, 3 and 4 times higher expression were 1 314, 157 and 19 respectively. The differences of signal pathways Genes obtained 18 statistically significant signal pathways, including closely related to cardiac development of signaling pathways, such as: Notch, PI3K / AKT, MAPK signaling pathway; random selection of expression of five genes were verified, the results showed that quantitative PCR Inspection results and chip screening results basically match. Conclusions The differentially expressed genes in cardiac muscle tissue of VSD and normal fetal hearts are related to the signal pathways (Notch, PI3K / AKT and MAPK signal pathways) that are closely related to cardiac development, which laid a good foundation for the study of the pathogenesis of congenital heart disease.