南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2013年
12期
1787-1791
,共5页
张珊珊%黎丽旋%黄载伟%辛小敏%肖冰
張珊珊%黎麗鏇%黃載偉%辛小敏%肖冰
장산산%려려선%황재위%신소민%초빙
CD133%CD200%大肠癌干细胞%基因芯片%MAPK信号通路
CD133%CD200%大腸癌榦細胞%基因芯片%MAPK信號通路
CD133%CD200%대장암간세포%기인심편%MAPK신호통로
CD133%CD200%colorectal cancer stem cells%genechip%MAPK signaling pathway
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。
目的:篩選CD133+CD200+大腸癌細胞併鑒定其相關基因的錶達。方法流式細胞術分選CD133+CD200+和CD133-CD200-大腸癌細胞,應用Affymetrix人類全基因組錶達譜芯片檢測這兩群細胞的基因錶達譜,初步篩選CD133+CD200+與CD133-CD200-大腸癌細胞之間的差異錶達基因,進而尋找與大腸癌榦細胞特異性相關的主效基因,最後利用qRT-PCR實驗對部分差異錶達基因進行驗證,以確定芯片檢測結果的可靠性。結果芯片結果顯示差異在3倍以上的基因共655箇,CD200+細胞錶達上調的基因290箇,錶達下調的基因共365箇,通過生物信息學分析和GENEMANIA共錶達構建,篩選齣3條(MDM2;PRKACG;CACNA1G)有特異相關的差異基因進行qRT-PCR驗證,結果與芯片結果完全相符。結論基因芯片技術通過篩選和鑒定CD133+CD200+大腸癌榦細胞相關基因,建立瞭特異性大腸癌榦細胞基因錶達譜,為大腸癌基因靶嚮治療及進一步研究提供依據。
목적:사선CD133+CD200+대장암세포병감정기상관기인적표체。방법류식세포술분선CD133+CD200+화CD133-CD200-대장암세포,응용Affymetrix인류전기인조표체보심편검측저량군세포적기인표체보,초보사선CD133+CD200+여CD133-CD200-대장암세포지간적차이표체기인,진이심조여대장암간세포특이성상관적주효기인,최후이용qRT-PCR실험대부분차이표체기인진행험증,이학정심편검측결과적가고성。결과심편결과현시차이재3배이상적기인공655개,CD200+세포표체상조적기인290개,표체하조적기인공365개,통과생물신식학분석화GENEMANIA공표체구건,사선출3조(MDM2;PRKACG;CACNA1G)유특이상관적차이기인진행qRT-PCR험증,결과여심편결과완전상부。결론기인심편기술통과사선화감정CD133+CD200+대장암간세포상관기인,건립료특이성대장암간세포기인표체보,위대장암기인파향치료급진일보연구제공의거。
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.