批次和流加培养中国仓鼠卵巢工程细胞的细胞周期相关基因转录谱分析  

Analysis of the transcriptional profiling of cell cycle regulatory networks of recombinant Chinese hamster ovary cells in batch and fed-batch cultures

作　　者：刘兴茂[1] 叶玲玲[1] 刘红[1] 李世崇[1] 王启伟[1] 吴本传[1] 陈昭烈[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物工程学报》2011年第8期1198-1205,共8页Chinese Journal of Biotechnology

基　　金："重大新药创制"科技重大专项资助课题(No.2009ZX09503-011)资助~~

摘　　要：以表达人重组尿激酶原中国仓鼠卵巢(CHO)工程细胞系11G-S为研究对象,运用基因芯片技术比较了CHO工程细胞在批次及流加培养不同生长阶段基因表达水平的差异,在此基础上采用Genmapp软件,同时结合已知的细胞周期信号通路图,着重分析了批次及流加培养CHO工程细胞的细胞周期调控基因转录谱差异。在基因芯片涉及的19 191个目标基因中,批次和流加培养不同生长阶段CHO工程细胞的下调表达的基因数量多于上调表达基因数目;两种培养模式下的基因差异表达有着明显的不同,尤其是在细胞生长的衰退期,流加培养CHO工程细胞中下调表达的基因数量明显多于批次培养。有关调控细胞周期关键基因的转录谱分析表明,CHO工程细胞主要是通过下调表达CDKs、Cyclin及CKI家族中的Cdk6、Cdk2、Cdc2a、Ccne1、Ccne2基因及上调表达Smad4基因,来达到调控细胞增殖及维持自身活力的目的。In the light of Chinese hamster ovary（CHO） cell line 11G-S expressing human recombinant pro-urokinase,the differences of gene expression levels of the cells in different growth phases in both batch and fed-batch cultures were revealed by using gene chip technology.Then,based on the known cell cycle regulatory networks,the transcriptional profiling of the cell cycle regulatory networks of the cells in batch and fed-batch cultures was analyzed by using Genmapp software.Among the approximate 19 191 target genes in gene chip,the number of down-regulated genes was more than those of up-regulated genes of the cells in both batch and fed-batch cultures.The number of down-regulated genes of the cells in the recession phase in fed-batch culture was much more than that of the cells in batch culture.Comparative transcriptional analysis of the key cell cycle regulatory genes of the cells in both culture modes indicated that the cell proliferation and cell viability of the cells in both batch and fed-batch cultures were mainly regulated through down-regulating Cdk6,Cdk2,Cdc2a,Ccne1,Ccne2 genes of CDKs,Cyclin and CKI family and up-regulating Smad4 gene.

关 键 词：中国仓鼠卵巢工程细胞 细胞培养 细胞周期 基因 转录谱 

分 类 号：Q813[生物学—生物工程]