基因芯片技术筛选和鉴定CD133^+CD200^+大肠癌干细胞相关基因  

Application of genome-wide genechip for screening and identifying genes related to CD133^+CD200^+ colorectal cancer stem cells

作　　者：张珊珊[1] 黎丽旋[1] 黄载伟 辛小敏[1] 肖冰[1] 

机构地区：[1]南方医科大学南方医院消化科//广东省胃肠疾病重点实验室,广东广州510515

出　　处：《南方医科大学学报》2013年第12期1787-1791,共5页Journal of Southern Medical University

基　　金：广东省科技计划项目(2010B031600097);卫生事业经费(粤财社[2011]315号)

摘　　要：目的筛选CD133+CD200+大肠癌细胞并鉴定其相关基因的表达。方法流式细胞术分选CD133+CD200+和CD133-CD200-大肠癌细胞,应用Affymetrix人类全基因组表达谱芯片检测这两群细胞的基因表达谱,初步筛选CD133+CD200+与CD133-CD200-大肠癌细胞之间的差异表达基因,进而寻找与大肠癌干细胞特异性相关的主效基因,最后利用qRT-PCR实验对部分差异表达基因进行验证,以确定芯片检测结果的可靠性。结果芯片结果显示差异在3倍以上的基因共655个,CD200+细胞表达上调的基因290个,表达下调的基因共365个,通过生物信息学分析和GENEMANIA共表达构建,筛选出3条(MDM2;PRKACG;CACNA1G)有特异相关的差异基因进行qRT-PCR验证,结果与芯片结果完全相符。结论基因芯片技术通过筛选和鉴定CD133+CD200+大肠癌干细胞相关基因,建立了特异性大肠癌干细胞基因表达谱,为大肠癌基因靶向治疗及进一步研究提供依据。Objective To screen and identity genes related to CD133＋CD200＋ colorectal cancer stem cells. Methods The two subpopulations of colorectal cancer cells, namely CD133＋CD200＋ and CD133-CD200- cells, were sorted and verified by flow cytometry. The gene expression profiles of CD133 ＋CD200 ＋and CD133-CD200- colorectal cancer cells were examined using Affymetrix Human U133 Plus2.0 genome- wide genechip. The differentially expressed genes between the two cell subpopulations were analyzed to identify the genes responsible for the main effect in association with colorectal cancer stem cells. Real-time quantitative PCR was performed to confirm some of the differentially expressed genes identified by genechip. Results The genechip result showed that 655 genes were differentially expressed in CD133＋CD200＋colorectal cancer stem cells by at least 3 folds, including 290 up-regulated and 365 down-regulated ones. Bioinformatics analysis and gene co-expression network building identified 3 genes （MDM2, PRKACG, and CACNA1G） with specific expression in CD133＋CD200＋colorectal cancer stem cells, and this result was confirmed by real-time quantitative PCR analysis. Conclusion A specific gene expression profile of colorectal cancer stem cells has been established through screening and identifying genes related to CD133＋CD200＋colorectal cancer stem cells by gene genechip technique, which provides a basis for further study of gene targeting therapy of colorectal cancer.

关 键 词：CD133 CD200 大肠癌干细胞 基因芯片 MAPK信号通路 

分 类 号：R735.34[医药卫生—肿瘤]