论文部分内容阅读
目的:从转录组水平识别弥漫型胃癌与正常胃黏膜间的基因表达差异,探讨胃癌分子发生、发展的机制。方法:收集22例弥漫型胃癌患者的胃癌组织及其相应远切端的正常胃黏膜。采用含14592个点的cDNA表达谱芯片建立胃癌基因表达谱。差异表达基因筛选标准为该基因在50%以上样本中肿瘤比正常组织荧光强度大2倍或以上(P<0.05)。采用系统聚类、方差分析(P<0.05)等方法进行差异表达分析,实时定量RT-PCR方法验证芯片结果。结果:胃癌组织与正常胃黏膜间的差异表达基因共357个,其中表达上调者153个;表达下调者204个。上调基因的功能主要与细胞骨架运动、基质重建和细胞黏附、细胞周期调控及信号传导相关;下调基因主要与消化功能、细胞免疫防御、代谢、凋亡抑制及电子传递相关。实时定量PCR验证结果与芯片结果一致。结论:运用cDNA芯片进行弥漫型胃癌基因表达谱分析,有助于从分子水平全方位理解弥漫型胃癌发病机制及生物学特性,也有助于发现新的分子诊断指标和基因治疗靶标。
OBJECTIVE: To identify the differences in gene expression between diffuse gastric cancer and normal gastric mucosa from the transcriptome level and to explore the molecular mechanism of gastric carcinogenesis. Methods: Twenty-two gastric cancer tissues and their corresponding normal gastric mucosa were collected from patients with diffuse gastric cancer. Gene expression profile of gastric cancer was established by cDNA microarray with 14,592 spots. The differential expression gene screening criteria for the gene in more than 50% of the samples of tumors than the normal fluorescence intensity of 2 times or more (P <0.05). Systemic analysis of variance and analysis of variance (P <0.05) were used for differential expression analysis, and real-time quantitative RT-PCR was used to verify the results of the chip. Results: There were 357 differentially expressed genes in gastric cancer tissues and normal gastric mucosa, among which 153 were up-regulated and 204 were down-regulated. The function of up-regulated genes is mainly related to cytoskeleton motility, matrix remodeling and cell adhesion, cell cycle regulation and signal transduction; down-regulated genes are mainly associated with digestive function, cellular immune defense, metabolism, apoptosis inhibition and electron transfer. Real-time quantitative PCR validation results consistent with the chip results. Conclusion: Gene expression profiling of diffuse gastric cancer using cDNA microarray is helpful to understand the pathogenesis and biological characteristics of diffuse gastric cancer from a molecular level. It is also helpful to discover new molecular diagnostic and gene therapy targets.