基因芯片研究尿道下裂的基因表达差异

Difference of genes expression in analyzed hypospadias by microarray

目的筛选出与尿道下裂发病相关的基因。方法尿道下裂病人的尿道或包皮组织,年龄配对的包皮过长患儿组织作为对照,提取组织mRNA,合成双股cDNA并纯化,再逆转录成cRNA,生物素标记的cRNA打成碎片后与人类基因芯片进行杂交,计算机工作站自动扫描并记录芯片上的信号强度,转化为EXCEL数据库,总共分析了22000个基因的表达差异。结果发现一组表达有显著差异的基因,包括CYR61,CTGF,ATF3和GADD45。结论这些表达差异的基因可能参与尿道下裂的发生和发展,为进一步研究关键基因提供了依据。

Objective To screen key genes evolved in hypospadias. Methods Total RNA were isolated from the tissues both come from hypospadias patients and age-matched phimosis as control. Double stranded cDNA was achieved from tissues mRNA and purified. An in vitro transcription reaction was performed to obtain biotin-labeled cRNA from the double-stranded cDNA. The fragmented biotin-labeled cRNA was hybridized to a human expression microarray. Signal intensity values of each element from chip data sets were scanned with the computer workstation automatically and analyzed with Affymetrix GeneChip analysis software （Microarray Suite）, then transformed to a Microsoft Excel data. The difference of 22 000 genes expression were analyzed. Results A number of significant differences in gene expression specifically with a group of genes, including CYR61, CTGF, ATF3, and GADD45 were screened. Clusters of genes showed profiles of up-regulation or down-regulation. Conclusion The findings provide profile of gene expression in hypospadias. Some of these genes contribute to the development of hypospadias. The data of gene expression in hypospadias is valuable for interested investigators to analysis and pick up key genes in hypospadias.