摘要
目的通过与人类全基因组寡核苷酸微阵列杂交及分析,筛查季节性变应性鼻炎(seasonal allergic rhinitis,SAR)与 SAR 合并哮喘患者鼻黏膜的差异基因表达谱。方法分别取5例SAR 及4例 SAR 合并哮喘患者的下鼻甲黏膜,提取总 RNA,合成生物素标记的 cRNA 探针,与美国Affymetrix 公司人类全基因组寡核苷酸微阵列 HG-U133-plus2杂交,并以反转录聚合酶链反应(reversetranscription polymerse chian reaction,RT-PCR)验证芯片结果。采用 GeneSpring7.3软件分析基因表达数据,对差异基因进行基因本体论分类和生物通路注释。结果在38 500多个基因(47 000个转录本)中,SAR 合并哮喘者,其鼻黏膜发生2倍以上差异表达的基因共有1 900个,其中849个基因表达上调,1 051个表达下调。这些基因功能主要与细胞代谢、基因转录、细胞增殖、信号转导、免疫应答、多种酶活性、膜受体结合活性、细胞结构蛋白活性等有关。生物通路分成161类,其中包含20个基因以上的通路有:细胞因子受体间相互作用,局部黏附,细胞黏附分子,细胞骨架肌动蛋白调节,细胞间信号传递,缝隙连接,促分裂原活化蛋白激酶信号传导通路,钙相关信号传导通路,白细胞跨膜转运以及嘌呤代谢等生物通路。结论 SAR 及 SAR 合并哮喘存在多基因表达调控的改变,对其差异表达基因的研究有助于阐明变应性鼻炎及其合并哮喘的发生发展机制。
Objective To screen the differential expression gene profile in nasal mucosa of seasonal allergic rhinitis（ SAR ） and SAR with asthma, oligonucleotide microarray （ Affymetrix HG-U133-plus2 ） was employed to analyze the changes of gene expressions with GeneSpring software. Methods Inferior turbinate mucosa was obtained from five SAR patients and four SAR with asthma patients. Total RNA was extracted from the nasal mucosal biopsies and pooled into one SAR control pool and one SAR with asthma patient pool, and biotin-labeled cRNA probes were hybridized with Affymetrix HG-U133-plus2 array. The hybridization results were confirmed by RT-PCR analysis . The analysis of differential expression profiles were performed by GeneSpring software 7.3. Results Out of 47 000 analysed transcripts, 1 900 genes were differentially expressed at least 2-fold in which 849 genes were up-regulated and 1 051 genes were down-regulated in nasal mucosa of SAR with asthma patients compared with that in SAR patients. These genes were involved in cell metabolism, gene transcription, cell proliferation, signal transduction, immune response, enzyme activity, transmembrane receptor activity, cytoskeletal protein binding, and many other aspects. Pathway analysis displayed 161 groups, of which including more than 20 genes were as follow： cytokine-cytokine receptor interaction, focal adhesion, cell adhesion molecules （CAMs）, regulation of actin cytoskeleton, cell communication, gap junction, MAPK signaling pathway, calcium signaling pathway, leukocyte transendothelial migration, and purine metabolism. Conclusions The data suggested that multigenetic expression and regulation changes were involved in the development of SAR and SAR complicated with asthma,whose molecular mechanisms might be elucidated by identification of these differential genes.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2007年第9期654-659,共6页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金
国家自然科学基金(30350006)
山西省资助出国留学人员基金(2005032)
关键词
鼻炎
变应性
季节性
哮喘
寡核苷酸序列分析
基因表达谱
Rhinitis, allergic, seasonal
Asthma
Oligonucleotide array sequence analysis
Gene expression profiling
