E2F-1过表达对胃癌细胞生物学行为的影响  

The molecular mechanism of the effects of E2F-1 overexpression on biological characteristics of human gastric carcinoma cell line

作　　者：王晓通[1] 李雷[1] 肖强[1] 谢玉波[2] 王长青[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学第一附属医院麻醉科,南宁530021

出　　处：《广东医学》2009年第11期1600-1603,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30860273)

摘　　要：目的探讨E2F-1对胃癌细胞生物学行为影响的分子机制。方法分别抽取转染E2F-1的胃癌MGC-803细胞和未转染E2F-1的胃癌MGC-803细胞的总RNA。纯化mRNA,逆转录合成cDNA标记后,与22K人类基因表达谱芯片进行杂交,扫描后筛选出表达差异的基因。随机选取3个差异表达基因,分别设计引物,用半定量RT-PCR检测验证。结果在21522条基因中,两组细胞间的差异表达基因有740条,其中上调基因405条,下调基因335条。差异基因中有73条功能信息不明。随机选取的3个差异表达基因的RT-PCR结果与基因芯片扫描结果相似,具有相同的方向性。结论胃癌的发生发展是多基因相互作用、多种信号通路相互调节的结果。生物信息学分析显示,E2F-1基因可以通过TP53信号传导途径影响胃癌MGC-803细胞生物学行为的改变,还与众多基因的差异性表达密切相关。Objective To study the molecular mechanism of the effects of E2F - 1 overexpression on biological characteristics of human gastric carcinoma cell line with eDNA microarray technique. Methods Total RNA were extracted from gastric cancer cell line MGC - 803 transfected with E2F - 1 and then purified. The cDNA obtained by reverse transcription polymerase chain reaction （RT- PCR）, was labeled with Cy5 and Cy3 fluorescence as probes, and then hy- bridized with gene chip containing 21522 human 22K gene expression profile. Subsequently, the two signal images were scanned by LuxScan 10K/A dual pathways laser scanner and analyzed by LuxScan3.0 image analysis software. We randomly chose three of the differentially expressed genes to confirm the array results using semi - quantitative RT - PCR. Results Of the 21522 target genes, 740 genes were screened out for differences in gene expression level. 405 of the 740 genes were up - regulated and 335 were down - regulated. 73 of differentially expressed genes were with unknown func- tion. The results of RT - PCR were well coincided with the cDNA microarray results. Conclusion The carcinogenesis of gastric carcinoma includes the interaction of multiple genes and regulation of many signal transduction pathways. E2F - 1 can affect the biological characteristics of MGC -803 by TP53 signal pathway, and is closely associated with the differentially expressed genes.

关 键 词：胃癌 E2F-1 基因表达谱芯片 差异表达基因 

分 类 号：R285.5[医药卫生—中药学] R737.9[医药卫生—中医学]