CAJ | 학술논문

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。
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.