CAJ | 학술논문

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.
목적 탐토인제대간충질간세포(hMSCs)화식관암세포융합전후기인표체적차이화신호통로적개변.방법 분선화감정융합세포(EMFs)후,통과기인심편비교식관암세포、hMSCs화EMFs적표체보.PCA취류분석전록조적정체관계,LIMMA분석획득차이표체기인,PCC학정차이표체기인적표체모식,재통과DAVID、ToppGene화MSigDB수거고분석각개모식적기인공능、신호통로부집、염색체정위화부집,이KEGGanim화Idiographica분별회제핵심차이표체기인적신호통로관련도화염색체분포도.결과 EMFs세포중DNA손상수복、세포주기조체화조망통로적핵심기인고표체,위EC9706혹hMSCs세포적4배(Pi ＜0.05),이억제인자DUSP6적고표체가능부반궤지억제촉분렬소원활화단백격매통로.hMSCs세포중세포인자화추화인자고표체,위EC9706혹EMFs세포적2배(Pi＜ 0.05),이EMFs세포중33개면역공능상관기인고표체,위EC9706세포적2배(Pi＜0.05).M기조체화안기산대사상관적위우식관암염색체확증구적39개차이표체기인중,기중30개표체하강,위EC9706세포적0.5배(Pi＜0.05).결론 hMSCs화EC9706세포융합후가능통과유도촉조망신호화DUSP6부반궤억제궤제래발휘억제식관암세포적작용,면역조절화분화상관기인적개변제시융합세포계승료간세포적부분면역공능.
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.