Seto N O, Seligy V L
Immunology Group, Institute of Biological Sciences, National Research Council of Canada, Ottawa, Ontario.
Cell Mol Biol (Noisy-le-grand). 1999 Mar;45(2):203-9.
The HT29 colonic carcinoma cell line has proven to be a very practical tool for modelling aspects of colonic cell differentiation and toxification by chemotherapeutic agents. As an approach to subclone and clarify molecular events involved in sublineage maturation, non-differentiated HT29 cells were electroporated with a dominant marker gene (NeoR) to convey aminoglycoside resistance (G418R). Transfectants surviving passage in glucose-G418 medium were >200 times the abundance of transient G418R cells of controls. Genomic analysis showed that each clonal type was unique in NeoR integration pattern while mitochondrial DNA copy was relatively unchanged. All of the randomly generated NeoR clones resembled the parental phenotype, but some over-produced the mucin, secretory cell type or the cell death phenotype after culturing in 2 mM sodium butyrate medium. Re-exposure to glucose medium restored the parental-like phenotype.
事实证明,HT29结肠癌细胞系是模拟结肠细胞分化和化疗药物毒性作用方面的一种非常实用的工具。作为亚克隆和阐明亚系成熟过程中涉及的分子事件的一种方法,用显性标记基因(NeoR)对未分化的HT29细胞进行电穿孔,以赋予氨基糖苷抗性(G418R)。在葡萄糖-G418培养基中传代存活的转染子数量是对照瞬时G418R细胞数量的200倍以上。基因组分析表明,每种克隆类型在NeoR整合模式上都是独特的,而线粒体DNA拷贝数相对不变。所有随机产生的NeoR克隆都与亲本表型相似,但在2 mM丁酸钠培养基中培养后,一些克隆过量产生粘蛋白、分泌细胞类型或细胞死亡表型。重新暴露于葡萄糖培养基后恢复了亲本表型。