摘要
目的 通过比较肝内胆管细胞癌RBE细胞株中NCAM＋ c-Kit＋与NCAM-c-Kit-的全基因组表达差异,筛选出与干细胞信号通路相关的差异基因,进一步揭示肝内胆管癌发生、发展的内在分子机制.方法 培养RBE细胞株,用免疫磁珠分选出NCAM＋ c-Kit＋ RBE与NCAM-c-Kit-RBE亚群,采用Agilent人全基因组表达谱芯片对NCAM＋ c-Kit＋ RBE与NCAM-c-Kit-RBE进行检测.通过SAS系统分析,得到两者干细胞信号通路的差异表达基因,进一步采用实时-PCR技术验证芯片结果.结果 芯片筛选出NCAM＋ c-Kit＋ RBE与NCAM-c-Kit-RBE亚群之间的差异基因共7270条[变化倍数（fc）≥2或fc≤0.5].与NCAM-c-Kit-RBE相比,NCAM＋ c-Kit＋ RBE上调基因3572条,下调基因3698条.其中干细胞信号通路相关差异基因共421条（fc≥2或fc≤0.5）,上调基因231条,下调基因190条.结论 高通量芯片能够筛选出NCAM＋ c-Kit＋ RBE与NCAM-c-Kit-RBE大量的差异基因.SAS分析系统能进一步分析与干细胞信号通路相关的差异基因.
Objective To analyze the differentially expressed genes between the NCAM ＋ c-Kit ＋RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma （ICC） cell lines,and to screen out the differentially expressed genes that are related to the stem cell signaling pathways.Methods Magnetic activated cell sorting was used to isolate the NCAM ＋ c-Kit ＋/NCAM-c-Kit-subset cells,and then Agilent Whole Human Genome Microarray Kit was used to test the difference in gene expressions between the NCAM ＋ cKit ＋ and NCAM-c-Kit-subset cells.The difference in gene expressions related to the stem cell signaling pathways was analyzed by the SAS system.The result of the microarray was further confirmed by RT-PCR.Results The total differentially expressed genes which could be found through gene microarray were 7270 [foldchange（fc） ≥2 or fc ≤0.5].Compared with the NCAM-c-Kit-RBE,3572 genes were upregulated while 3698 genes were downregulated.The differences in gene expressions related to the stem cell signaling pathways were 421 （fc ≥2 or fc ≤ 0.5）,among which 231 genes were upregulated while 190 genes were downregulated.Conclusions High-flux microarray could be used to screen out lots of differentially expressed genes between the NCAM ＋ c-Kit ＋ and NCAM-c-Kit-RBE cells.The differences in gene expression in the stem cell signaling pathways could also be further analyzed using the SAS system.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2015年第6期373-376,共4页
Chinese Journal of Hepatobiliary Surgery
基金
山西省回国留学人员科研项目（2011-046）
山西省留学回国人员科技活动择优资助项目资助（2011-762）
山西医科大学基础医学院331基础医学科技培植基金计划项目（201407）
山西医科大学校创新基金（01201302）
关键词
肝内胆管癌
肝脏干/祖细胞
基因芯片
信号通路
Intrahepatic cholangiocarcinoma
Hepatic stem/progenitor cell
Gene microarray
Signaling pathway
