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目的:观察法乐四联症(TOF)相关基因的表达变化。方法:使用Biostar 80s型人类基因表达谱微阵列初步筛选TOF和室间隔缺损患者右心室心肌的差异表达基因。结合初筛实验、网上孟德尔人类遗传数据库及心脏基因表达数据库的查询结果,遴选补充至500条目标基因。应用3张定制的1000点人类基因微阵列,双重点样,分别筛选TOF(组)25例、不伴有先天心脏畸形的引产胎儿(FETAL组)5例、右室双出口(组)5例、室间隔缺损(组)15例患者,检测右心室流出道肌肉中的差异表达基因,进行生物信息学分析。采用实时荧光定量聚合酶链反应技术验证微阵列结果。结果:用Biostar 80s微阵列发现186条差异表达基因,用定制微阵列则分别发现104条(TOF/FETAL)、13条(TOF/右室双出口)和315条(TOF/室间隔缺损)。剔除结果可能不可靠者(179条),根据差异表达特征,将剩余基因归入14个相关基因群。实时荧光定量聚合酶链反应证实,胰岛素样生长因子结合蛋白3、Ⅰ型胶原α_2链蛋白与Ⅲ型胶原α_1链蛋白3个基因的差异表达与微阵列结果相一致。结论:应用基因微阵列技术进行高通量并行分析,可确定与TOF相关的基因表达谱。胶原家族成员及胰岛素样生长因子结合蛋白3可能与TOF的发生发展过程密切相关。
Objective: To observe the changes of gene associated with tetralogy of Fallot (TOF). Methods: Biostar 80s human gene expression microarray was used to screen differentially expressed genes in right ventricular myocardium of patients with TOF and ventricular septal defect. Combined with screening test, online Mendel human genetic database and cardiac gene expression database query results, the selection of up to 500 target genes. Three custom-made 1,000-point human gene microarrays were double-spotted and were screened for 25 cases of TOF, 5 cases of FETAL without congenital heart malformation, 5 cases of right ventricular double outlet (group) , Ventricular septal defect (group) 15 patients, right ventricular outflow tract muscle in the differential expression of genes for bioinformatics analysis. Microarray results were validated using real-time fluorescence quantitative polymerase chain reaction. RESULTS: A total of 186 differentially expressed genes were found using the Biostar 80s microarray. Of 104 (TOF / FETAL), 13 (TOF / RV extrinsic) and 315 (TOF / ventricular septal defects) were detected with a custom microarray. Excluding the results may be unreliable (179), according to the characteristics of differential expression, the remaining genes included in 14 related gene groups. Real-time fluorescence quantitative polymerase chain reaction confirmed that the differential expression of insulin-like growth factor binding protein 3, type I collagen alpha 2 chain protein and type III collagen alpha 1 chain protein 3 genes consistent with microarray results. Conclusions: High-throughput parallel analysis using gene microarray technology can identify gene expression profiles associated with TOF. Collagen family members and insulin-like growth factor binding protein 3 may be closely related to the occurrence and development of TOF.