南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (12): 1787-.

• • 上一篇    下一篇

基因芯片技术筛选和鉴定CD133+CD200+大肠癌干细胞相关基因

张珊珊,黎丽旋,黄载伟,辛小敏,肖冰   

  • 出版日期:2013-12-20 发布日期:2013-12-20

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

  • Online:2013-12-20 Published:2013-12-20

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。

Abstract: 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.