实时荧光定量RT-PCR和寡核苷酸芯片方法分析食管腺癌组织中基因的表达

Gene expression in esophageal adenocarcinoma by oligomicroarray and real time RT-PCR

目的采用实时荧光定量RT-PCR和寡核苷酸芯片技术分析食管腺癌基因表达的特征。方法应用寡核苷酸芯片筛选6例食管腺癌基因表达谱,实时荧光定量RT-PCR验证其中8个基因的表达变化。结果2倍差异表达基因(DEGs)中,上调基因共212个,下调基因共126个;涉及细胞周期相关基因、信号转导相关基因、血管生成相关基因、细胞增殖相关基因、凋亡抑制基因、抑癌基因、黏附和代谢等。实时荧光定量RT-PCR和寡核苷酸芯片技术对8个基因的检测结果一致,不仅变化方向相同,而且表达值非常接近。结论食管腺癌的发生、发展涉及多基因多步骤,是一个复杂的过程。寡核苷酸芯片和实时荧光定量RT-PCR分析基因表达变化都是可信的,基因芯片筛选基因表达谱具有高通量大规模的特点,而实时荧光定量RT-PCR则适合单个基因表达变化的研究,两者互为补充和印证。

Objective To analyze the gene expression by real time RT-PCR and oligomicroarray in the human esophageal adenocarcinoma （EAC）. Methods The gene expression profile was screened by oligomicroarray in 6 cases of EAC, and 8 genes of which were confirmed by real time RT-PCR. Results There were 212 up-regulated genes and 126 down-regulated genes among 2-fold DEGs, including cell cycle-related genes, signal transduction-related genes, angio- genesis-related genes, cell proliferation-related genes, metastasis-related genes, apoptosis inhibiting genes, tumor-sup- pressing genes and so on. The identical direction and almost identical genes expression of mRNA detected by real time RT-PCR were accorded with those by oligomicroarray. Conclusion The development and progression of EAC is a complicated process involving multigenes and multiprocedures. Oligomicroarray and real time RT-PCR techniques are faithful in the analysis of genes expression. Oligomicroarray possesses high-throughput characteristic and real time RT-PCR is applicable to the analysis of target gene expression.