原发性肾病综合征的分子肖像(英文)

Molecular portrait of primary nephrotic syndrome

摘要:目的比较原发性肾病综合征患儿和健康儿童外周血单个核细胞基因表达谱的差异,筛选原发性肾病综合征相关基因,绘制原发性肾病综合征的分子肖像,系统阐明原发性肾病综合征发生的分子机制。方法分离6例原发性肾病综合征患儿和6例与之匹配的健康儿童的外周血单个核细胞,用TRIzol一步法提取外周血单个核细胞的总RNA,再用逆转录-PCR法以Cy5和Cy3分别标记原发性肾病综合征患儿和健康儿童的mRNA而成为cDNA探针。将两种探针混合后与点有8096条基因的芯片杂交,经严格洗涤后用GenePix4000B扫描仪扫描杂交信号,用GenePixPro3.0软件分析扫描结果。结果8096条被检基因中共有418条在PNS患儿和健康儿童的PBMC中存在差异表达,其中128条基因在PNS患儿的PBMC中表达升高,290条基因表达降低,102条基因在本研究的6对PNS患儿和健康儿童配伍对照中均存在差异表达。这102条基因根据功能可分为10类:免疫相关基因、细胞骨架相关基因、信号转录相关基因、细胞周期及增殖凋亡相关基因、癌基因和抑癌基因、基因复制和表达相关基因、离子通道和转运蛋白相关基因、受体相关基因、代谢相关基因及其它功能尚不清楚的基因。结论原发性肾病综合征的发生、发展是许多相互作用的基因共同作用的结果。

To compare the gene expression profile between peripheral blood mononuclear cell (PBMC) of children with primary nephrotic syndrome (PNS) and healthy control, screen the candidate primary nephrotic syndrome associated genes, make the molecular portrait of primary nephrotic syndrome, elucidate the molecular mechanism of primary nephrotic syndrome systemically. PBMCs were separated from 6 children with PNS and 6 healthy controls. Total RNA was isolated from PBMCs with TRIzol single-step method, mRNA from children with PNS and healthy controls were labelled with Cy5 and Cy3 separately by reverse transcription-PCR(RT-PCR)as cDNA probes. The two kinds of probe were mixed and hybridized with gene chip which contain 8096 cDNA clones.After high-strigent washing, GenePix 4000B was used for scanning the hybridizing signals and GenePix Pro 3.0 for data analysis. Of the 8096 genes monitored, 418 genes showed differential expression between children with PNS and healthy controls, in which 128 genes was up-regulated and 290 genes was down-regulated in PBMCs of patients with PNS, 102 genes showed differential expression between all the 6 pairs of children with PNS and their matched healthy control. The 102 genes can be divided into 10 categories on the basis of their function: immune-associated genes, cytoskeleton-associated genes, signal transduction-associated genes,cell cycle, proliferation,apoptosis-associated genes, proto-oncogenes and tumor suppressor genes,gene replication and expression-associated genes, channel protein and transporter-associated genes, receptor-associated genes, metabolism-associated genes and other genes with undetermined function. [Conclusion] The onset and progression of primary nephrotic syndrome is the result of the action of many interactive genes.