EVALUATION OF DOMINANTLY-ACTING RAS ONCOGENES

描述：本提案的目的是确定维护是否 人类癌细胞的致瘤表型取决于持续的 存在激活的ras癌基因。三种不同的实验 协议被提出：1)激活的ras等位基因将被灭活 靶向同源重组，或通过双重表达 突变的ras显性负等位基因；2)显性负性等位基因 两个特征良好的ras信号通路中的蛋白质将是 用于选择性地干扰每一条通路；以及3)ras反义 寡核苷酸将用于修饰体外生长特性 和含有活化的ras的人肿瘤细胞的致瘤能力 等位基因。用于这些实验的人类肿瘤细胞是 激活H-、K-或N-ras突变的杂合子。化验分析 转基因或寡核苷酸处理的细胞将包括 Ras和ras依赖的信号通路的生化检查， 体外生长和形态观察，小鼠肿瘤形成。

DESCRIPTION: The aim of this proposal is to determine whether maintenance of the human cancer cell tumorigenic phenotype depends on the continued presence of activated ras oncogenes. Three different experimental protocols are proposed: 1) activated ras alleles will be inactivated by targeted homologous recombination, or by the expression of a doubly mutant ras dominant negative allele; 2) dominant negative alleles of proteins in the 2 well characterized ras signalling pathways will be used to selectively disrupt each pathway; and 3) ras antisense oligonucleotides will be used to modify the in vitro growth properties and tumorigenic potential of human tumor cells containing activated ras alleles. The human tumor cells used for these experiments are heterozygous for activating mutations in H-, K- or N-ras. Assays of genetically modified or oligonucleotide-treated cells will include biochemical examination of ras and ras-dependent signalling pathways, growth and morphology in vitro, and tumor formation in mice.