中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2015年
6期
373-376
,共4页
王婕%徐菁%贺杰峰%李灵敏%覃艳红%陈秀花%贺艳玲
王婕%徐菁%賀傑峰%李靈敏%覃豔紅%陳秀花%賀豔玲
왕첩%서정%하걸봉%리령민%담염홍%진수화%하염령
肝内胆管癌%肝脏干/祖细胞%基因芯片%信号通路
肝內膽管癌%肝髒榦/祖細胞%基因芯片%信號通路
간내담관암%간장간/조세포%기인심편%신호통로
Intrahepatic cholangiocarcinoma%Hepatic stem/progenitor cell%Gene microarray%Signaling pathway
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.
目的 通過比較肝內膽管細胞癌RBE細胞株中NCAM+ c-Kit+與NCAM-c-Kit-的全基因組錶達差異,篩選齣與榦細胞信號通路相關的差異基因,進一步揭示肝內膽管癌髮生、髮展的內在分子機製.方法 培養RBE細胞株,用免疫磁珠分選齣NCAM+ c-Kit+ RBE與NCAM-c-Kit-RBE亞群,採用Agilent人全基因組錶達譜芯片對NCAM+ c-Kit+ RBE與NCAM-c-Kit-RBE進行檢測.通過SAS繫統分析,得到兩者榦細胞信號通路的差異錶達基因,進一步採用實時-PCR技術驗證芯片結果.結果 芯片篩選齣NCAM+ c-Kit+ RBE與NCAM-c-Kit-RBE亞群之間的差異基因共7270條[變化倍數(fc)≥2或fc≤0.5].與NCAM-c-Kit-RBE相比,NCAM+ c-Kit+ RBE上調基因3572條,下調基因3698條.其中榦細胞信號通路相關差異基因共421條(fc≥2或fc≤0.5),上調基因231條,下調基因190條.結論 高通量芯片能夠篩選齣NCAM+ c-Kit+ RBE與NCAM-c-Kit-RBE大量的差異基因.SAS分析繫統能進一步分析與榦細胞信號通路相關的差異基因.
목적 통과비교간내담관세포암RBE세포주중NCAM+ c-Kit+여NCAM-c-Kit-적전기인조표체차이,사선출여간세포신호통로상관적차이기인,진일보게시간내담관암발생、발전적내재분자궤제.방법 배양RBE세포주,용면역자주분선출NCAM+ c-Kit+ RBE여NCAM-c-Kit-RBE아군,채용Agilent인전기인조표체보심편대NCAM+ c-Kit+ RBE여NCAM-c-Kit-RBE진행검측.통과SAS계통분석,득도량자간세포신호통로적차이표체기인,진일보채용실시-PCR기술험증심편결과.결과 심편사선출NCAM+ c-Kit+ RBE여NCAM-c-Kit-RBE아군지간적차이기인공7270조[변화배수(fc)≥2혹fc≤0.5].여NCAM-c-Kit-RBE상비,NCAM+ c-Kit+ RBE상조기인3572조,하조기인3698조.기중간세포신호통로상관차이기인공421조(fc≥2혹fc≤0.5),상조기인231조,하조기인190조.결론 고통량심편능구사선출NCAM+ c-Kit+ RBE여NCAM-c-Kit-RBE대량적차이기인.SAS분석계통능진일보분석여간세포신호통로상관적차이기인.
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.
