cDNA芯片研发及其在肥胖患者脂肪基因差异表达研究中的应用

Development of High-density cDNA Microarray and Its Application in The Study of Differential Gene Expression Profile of Adipose Tissue From Obese Patients

为了加快基因功能的研究 ,利用已有的来源于不同组织的cDNA克隆 ,并通过交换和购买补充了低丰度和染色体覆盖不完全的部分cDNA ,研制开发出具有相当代表性、覆盖较完全的高密度cDNA表达型基因芯片 ,每张芯片上含有 384个质控DNA和 12 6 30个cDNA探针 ,其中包括 12 5 0 8个Unigene和 12 2个表达序列标签 (EST) .利用这些芯片 ,对肥胖患者及正常人内脏脂肪组织基因表达谱进行了初步研究 ,并发现在肥胖患者内脏脂肪组织差异表达的基因 ,其中上调的有与凋亡相关的基因、与免疫有关的基因以及与能量代谢有关的基因等 ,而下调的主要是与脂肪酸及胆固醇合成有关的基因 。

To accelerate study of gene function, a home-made high-density cDNA microarray was developed, using the cDNA clones from different types of tissues. In order to cover as far as possible the whole genome, cDNA clones that essentially represented genes with low abundance were introduced from Resgene Company. Totally 384 quality control DNA and 12 630 cDNA fragments, which includes 12 508 Unigenes and 122 ESTs, were spotted on the chip, after optimization. The home-made cDNA microarray was evaluated with Microarray ScoreCard and proved to be representative and reliable. These microarrays were then used to study the differential gene expression profiles of visceral adipose tissues between obese patients and normal subjects. The results showed that genes involved in immune-regulation, apoptosis and metabolism of energy were up-regulated in obese patients, while genes involved in lipid synthesis were down-regulated in obese patients. Further exploration of these genes will greatly improve the understanding of the pathogenesis of obesity.