基因表达谱分析小鼠膀胱癌VEGF信号转导通路的作用  

Microarray analysis on the role of VEGF signaling pathway for the growth and metastasis of murine bladder carcinoma

作　　者：王丰[1] 田毓华[1] 陈霞芳[1] 黄倩[1] 

机构地区：[1]上海交通大学附属第一人民医院中心实验室,上海200080

出　　处：《肿瘤》2007年第11期847-852,共6页Tumor

基　　金：上海市卫生局百人计划资助项目(编号:99BR006);国家自然科学基金杰出青年项目(编号:30325043);国家重点基础研究计划(编号:2004CB518804);上海市青年科技启明星计划(编号:04QMX1417)

摘　　要：目的:探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)在肿瘤新生血管形成过程中的作用,与肿瘤生长、转移的关系及其作用机制。方法:通过转染反义VEGF表达质粒,建立遗传背景相同但产生VEGF能力明显不同的肿瘤细胞株,观察VEGF对细胞生长速率、体内成瘤及肿瘤生长和转移能力的影响。分别将来自高与低水平表达VEGF的肿瘤细胞和肿瘤组织的cDNA与含有16 361个人cDNA芯片杂交,分析差异表达基因及基因表达谱变化,并通过实时荧光定量PCR进行验证。结果:反义VEGF表达质粒可选择性抑制肿瘤细胞产生VEGF,VEGF产生减少并不影响肿瘤细胞在体外的生长速率和生长方式,但在小鼠体内生长明显变慢,转移减少。基因芯片杂交结果显示高与低水平表达VEGF的肿瘤细胞和肿瘤组织中存在82个共同的差异表达在2倍以上的基因。进一步从中选择出23个杂交信号强度较高(信号强度大于或等于40)的基因,根据cDNA芯片上的探针顺序,比对小鼠cDNA及EST数据库,在其中杂交信号强度较高(信号强度大于或等于40)的23个基因中,找到了17个同源基因,其中9个为功能已知基因。由于这些基因并非已知的新生血管形成有关的基因,提示可能有更多的新基因参与肿瘤新生血管形成。结论:VEGF通过调控肿瘤新生血管形成影响肿瘤的生长和转移,而VEGF的作用需要多个基因或信号通路构成的复杂网络来完成。Objective： Angiogenesis has been indicated to play pivotal roles in both the initial development and metastasis of solid tumor. Among the numerous factors, vascular endothelial growth factor （VEGF） is widely recognized. This study aimed to discuss the role of VEGF in angiogenesis and its relationship with the growth and metastasis of murine bladder carcinoma as well as the action mechanism. Methods： The BTT-VEGFanti cells, which constitutively express the anti-sense VEGFI21 gene, were derived by stably transfecting the BTT-T739 cells with the anti-sense VEGF121 plasmid. The effects of VEGF on the growth rates, tumor formation capability, proliferation and metastasis of the two cells both in vitro and in vivo were observed. Isolated and purified mRNA from the two kinds of cells and tissues were reverse transcribed and hybridized with cDNA gene chips including 16,361 human genes. The differentiated gene expression was recorded and the changes in the gene expression profiles were analyzed. Real-time RT-PCR was used to identify the microarray results. Some genes with more than 2 fold changes were searched in iPath for discovering the related pathways. Results： Anti-sense VEGF expression plasmid selectively inhibited the production of VEGF by tumor cells. The declined level of VEGF had no effects on the growth rate and growth pattern of tumor cells in vitro. But the tumor growth was significantly slowed and metastasis was significantly inhibited in vivo after treatment with anti-sense VEGF expression plasmid. Microarray hybridization results showed that there existed 82 common genes with more than 2 fold differentiated expression in tumor cells and tumor tissues with high or low expression of VEGF. Twenty-three genes （ signaling density≥40） were further selected. Out of the 23 genes, 17 genes were found to be homo geneous by comparison with murine cDNA and EST database according to the probe sequence in cDNA microarray. The functions of 9 genes were already known. They were not known genes in induction

关 键 词：膀胱肿瘤 实验性 基因表达谱 血管内皮生长因子 

分 类 号：R737.14[医药卫生—肿瘤] R73-37[医药卫生—临床医学]