Barrett食管基因差异表达谱初步探讨

Preliminary Appraisal of Differential Expression Gene Profile in Barrett's Esophagus

背景:病变组织差异表达基因的分析有助于寻找与疾病发生、发展和临床治疗有关的关键分子。目的:应用寡核苷酸芯片对Barrett食管与正常食管黏膜组织的差异表达基因进行高通量分析,以期筛选与Barrett食管发生、发展相关的分子标记物。方法:取6例Barrett食管患者病变食管黏膜和相应正常食管黏膜组织,Trizol一步法抽提总RNA,纯化后合成cRNA探针;探针荧光标记和纯化后,将两种组织探针分别与Agilent全基因组寡核苷酸芯片(含30968个基因组探针)进行杂交;获取芯片扫描图谱,以特征提取软件行定量分析、处理。结果:Barrett食管与正常食管黏膜组织的2倍差异表达(Ratio值≥2或≤0.5)基因中,上调基因142个,下调基因284个,涉及细胞周期相关基因、信号转导相关基因、黏蛋白相关基因、癌基因和抑癌基因、骨形态蛋白基因、凋亡抑制基因、bcl-2家族相关基因等。结论:高通量的基因芯片技术可筛选出大量Barrett食管相关基因,对这些相关基因的功能进行验证将有助于找到Barrett食管发生、发展的关键基因或通路。

Differential expression gene profile analysis has made it possible to identify novel genes with altered expression in lesion tissues, which were relevant to the development, progression and clinical therapy of diseases. Aims： To analyze the differential expression genes between Barrett＇s esophagus and normal esophageal mucosal tissues by high- throughput oligonucleotide microarray, and to explore the potential molecular markers correlated with the development and progression of Barrett＇s esophagus. Methods： Total RNAs were isolated from 6 Barrett＇s esophagus and matched normal esophageal mueosal tissues using one-step Trizol method, cRNA probes were synthesized and then labeled with fluorescence and purified. Then the cRNA probes of both Barrett＇s esophagus and matched normal esophageal mueosal tissues were hybridized with Agilent whole genome oligonucleotide microarray （containing 30 968 genome probes）, respectively. The fluorescence intensity patterns were scanned and quantified by feature extraction software. Results： There were 142 up-regulated genes and 284 down-regulated genes with 2-fold differential expression （Ratio≥2 or ≤0.5） between Barrett＇s esophagus and matched normal esophageal mucosal tissues, including cell cycle-related genes, signal transduetion-related genes, mucin-re]ated genes, oncogenes and anti-oncogenes, bone morphogenetic protein genes, apoptosis-inhibiting genes and bcl-2 family-related genes, etc. Conclusions： Many Barrett＇s esophagus-associated genes can be screened by the high-throughput gene chip technique. Functional study of these genes may help identifying the key genes or pathways involved in the development and progression of Barrett＇ s esophagus.