人脐带间充质干细胞与食管癌细胞融合后的转录组比较  

A comparative analysis of the changes of transcriptome after fusion of esophageal cancer cells with human umbilical mesenchymal stem cells

作　　者：王勇[1] 周兵[2] 樊宏斌[1] 遇珑[1] 郭黎平[1] 陆士新[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤医院肿瘤研究所分子肿瘤学国家重点实验室病因及癌变研究室北京市癌症发生及预防分子机理重点实验室,100021 [2]中国科学院遗传与发育研究所发育及分子生物学重点实验室系统生物学研究中心

出　　处：《中华肿瘤杂志》2014年第10期726-732,共7页Chinese Journal of Oncology

基　　金：国家重点基础研究发展规划项目（2009CB521803）;国家高技术研究发展计划项目（2012AA02A503）

摘　　要：目的 探讨人脐带间充质干细胞（hMSCs）和食管癌细胞融合前后基因表达的差异和信号通路的改变.方法 分选和鉴定融合细胞（EMFs）后,通过基因芯片比较食管癌细胞、hMSCs和EMFs的表达谱.PCA聚类分析转录组的整体关系,LIMMA分析获得差异表达基因,PCC确定差异表达基因的表达模式,再通过DAVID、ToppGene和MSigDB数据库分析各个模式的基因功能、信号通路富集、染色体定位和富集,以KEGGanim和Idiographica分别绘制核心差异表达基因的信号通路关联图和染色体分布图.结果 EMFs细胞中DNA损伤修复、细胞周期阻滞和凋亡通路的核心基因高表达,为EC9706或hMSCs细胞的4倍（Pi ＜0.05）,而抑制因子DUSP6的高表达可能负反馈地抑制促分裂素原活化蛋白激酶通路.hMSCs细胞中细胞因子和趋化因子高表达,为EC9706或EMFs细胞的2倍（Pi＜ 0.05）,而EMFs细胞中33个免疫功能相关基因高表达,为EC9706细胞的2倍（Pi＜0.05）.M期阻滞和氨基酸代谢相关的位于食管癌染色体扩增区的39个差异表达基因中,其中30个表达下降,为EC9706细胞的0.5倍（Pi＜0.05）.结论 hMSCs和EC9706细胞融合后可能通过诱导促凋亡信号和DUSP6负反馈抑制机制来发挥抑制食管癌细胞的作用,免疫调节和分化相关基因的改变提示融合细胞继承了干细胞的部分免疫功能.Objective To compare the transcriptome of esophageal cancer cells （EC9706）,human mesenchymal stem cells （MSCs）,and after fusion of esophageal cancer cells with MSCs,and to further study their different expression profiles and the changes of their signaling pathways.Methods We examined the gene expression profiles of these cells with transcriptome microarray using LIMMA package and several web-based applications,such as DAVID,ToppGene and MSigDB.The resulting sets of differentially expressed genes （DEGs） were comprehensively analyzed to identify the pathways and their changes after the cell fusion.Results A total of 4 548 significantly DEGs among the three cell lines were found by LIMMA.Three functional annotation web tools predicted that DNA damage repair,cell cycle arrest and apoptosis pathways were enriched.Total DEGs were mapped to the canonic pathways with KEGGanim which depicted that the core genes of DNA damage repair,cell cycle arrest and pro-apoptosis were up-regulated in fusion cells,and they mightbe combined to respond the fusion-induced damage stress.The up-regulation of suppressive factor DUSP6 might feedback inhibit the MAPK signaling pathway in the fusion cells,too.Conclusions Transcriptome analysis suggests that hMSCs and EC9706 cell fusion may inhibit growth of EC cells by induction of pro-apoptotic signaling and DUSP6 negative feedback inhibition mechanism.In addition,the changes of immune regulation-related and differentiation-related genes indicate that the fusion cells inherited certain immune-suppressive function from the stem cells.

关 键 词：间质干细胞 脐带 食管肿瘤 细胞融合 转录因子 

分 类 号：R735.1[医药卫生—肿瘤]