CARCINOGEN-TRANSFORMED HUMAN CELLS, GENETIC TRAITS

本研究的两个基本目标是首先研究 特定致癌物修饰核苷酸之间的定量关系 在二倍体人类细胞基因组中，突变体（HGPRT） 位点）或锚定非依赖性细胞中观察到的存活细胞 第二，使用基本的细胞遗传学和基因转移技术 为了表征不依赖于贴壁的人类细胞克隆或紧急的 这些细胞系是通过我们的致癌物处理产生的。 我们将 确定这些克隆人类细胞的表型特征 群体（例如，作为细胞在软琼脂生长中的锚定独立性 系，其DNA转化NIH/3 T3细胞致瘤性的能力）是 与任何持续观察到的细胞遗传学变化相关，或 在未来的细胞杂交实验中的显性-隐性行为。 综合分析（HPLC），细胞（突变选择，软琼脂 克隆、中期核型分析）和分子（DNA 纯化/转染）技术用于这些实验。 我们 目的是整合核苷酸修饰的生物化学观察， 通过转化表型的分子研究进行突变诱导， 它的显性/隐性行为，以发展的理解， 肿瘤性转化

The two basic objectives of this research are first to study the quantitative relationship between specific carcinogen-modified nucleotides in a diploid human cell genome and the frequency with which mutant (HGPRT locus) or anchorage-independent cells are observed in the surviving cell progeny; and second, to use basic cytogenetic and gene transfer techniques to characterize the anchorage-independent human cell clones or emergent cell lines which are produced by our carcinogen treatment. We will determine whether the phenotypic traits in these cloned human cell populations (e.g. anchorage independence in soft agar growth as a cell line, ability of their DNA to transform NIH/3T3 cells tumorigenicity) are relatable to any consistently observed cytogenetic change, or dominant-recessive behavior in future cell hybridization experiments. Integrated analytical (HPLC), cellular (mutation selection, soft agar cloning , metaphase karyotyping) and molecular (DNA purification/transfection) techniques are used in these experiments. Our aim is to integrate biochemical observations of nucleotide modification and mutation induction with molecular studies of the transformed phenotype and its dominant/recessive behavior in order to develop an understanding of neoplastic transformation.