cDNA微阵列对22例弥漫型胃癌基因的检测  

Gene expression profiling of diffuse-type gastric cancer by cDNA microarray

作　　者：张晓青[1] 杨燕青[2] 刘炳亚[1] 金晓龙[3] 李炜[1] 唐凯玲[2] 张庆华[2] 林言箴[1] 朱正纲[1] 

机构地区：[1]上海第二医科大学附属瑞金医院外科消化外科研究所,200025 [2]生物芯片国家工程中心上海中心上海生物芯片有限公司 [3]上海第二医科大学附属瑞金医院病理科,200025

出　　处：《中华肿瘤杂志》2006年第2期116-119,共4页Chinese Journal of Oncology

摘　　要：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机制。方法收集22例弥漫型胃癌患者的新鲜冻存胃癌组织及同例相应正常胃黏膜。杂交芯片采用含14592个点的cDNA表达谱芯片。差异表达基因的筛选标准为该基因在50%以上样本中的肿瘤与正常组织荧光强度比(ratio比值)>2或<0.5。采用系统聚类法进行基因表达的相似性分析,标本组间比较采用方差分析。应用实时定量RT-PCR方法对芯片结果进行验证。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,下调者204个。上调基因功能主要与细胞骨架运动、基质重建、细胞增殖及信号传导等相关;下调基因功能则主要与细胞免疫防御、毒理代谢、功能分化、核-浆转运及凋亡抑制等相关。TNM分期的Ⅰ+Ⅱ期组与Ⅲ+Ⅳ期组间,有7个基因的表达差异有统计学意义(P<0.05)。RT-PCR验证结果与芯片表达结果一致。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阐明弥漫型胃癌的发病机制及生物学特性,也有助于进一步发现新的分子诊断指标和基因治疗靶标。Objective To identify cancer-related genes in diffuse-type gastric cancer and to explore its molecular mechanism by cDNA microarray analysis. Methods A total of 22 pairs of diffuse-type gastric cancer tissue and the corresponding normal mucosa were taken and freshly frozen, cDNA microarray with 14 592 genes/ESTs was used. Genes were considered to be up- or down-regulated when the fluorescent intensity ratio between tumor and normal mucosa was over 2-fold in over 50% of the samples （ P 〈 0.05 ）. Hierarchical clustering of regulated genes was performed as a measure to study expressional similarity. Validation of array results was carried out by real time quantitative PCR （QPCR）. Results Compared with those of corresponding normal mucosa, there were a total of 153 genes,ZESTs up-regulated and 204 downregulated in diffuse-type gastric cancer. Hierarchical clustering demonstrated that the genes belonging to the same subgroup displayed similar function. Most of the overexpressed genes were those related to cell adhesion, cell motility, matrix reconstruction, cell proliferation and/or signal transduction; while genes related to defense response, toxicoid metabolism, DNA repairing, nuclear-cytoplasmic transport and/or antiapoptosis made up the main list of the underexpressed genes. Seven genes showed higher expression in TNM （ TⅠ＋ TⅡ） group than in （ TⅢ＋ TⅣ） group . QPCR confirmed the array analysis results . Conclusion Gene expression profiling by cDNA microarray analysis provides not only molecular understanding of biological properties of cancer, but may also be helpful in discovering new diagnostic markers and therapeutic targets in gastric adenocarcinoma.

关 键 词：基因表达 胃癌 CDNA微阵列 

分 类 号：R735.2[医药卫生—肿瘤]