CARCINOGEN-TRANSFORMED HUMAN CELLS, GENETIC TRAITS

这项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以及突变(HGPRT)的频率 在存活的细胞中观察到)或与锚定无关的细胞 第二，使用基本的细胞遗传学和基因转移技术 描述与锚定无关的人类细胞克隆或新出现的 通过我们的致癌物治疗产生的细胞系。我们会 确定这些克隆的人类细胞的表型特征 群体(例如，软琼脂作为细胞生长的锚定独立性 它们的DNA转化NIH/3T3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有关，或 未来细胞杂交实验中的显性-隐性行为。 综合分析(高效液相色谱法)，细胞(突变选择，软琼脂 克隆、中期核型)和分子(DNA 纯化/转基因)技术在这些实验中被使用。我们的 目的是将核苷酸修饰的生物化学观察与 突变诱导与转化表型和分子生物学研究 它的显性/隐性行为，以发展对 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.