Kasid U, Pfeifer A, Weichselbaum R R, Dritschilo A, Mark G E
Science. 1987 Aug 28;237(4818):1039-41. doi: 10.1126/science.3616625.
In order to identify the genetic factors associated with the radiation-resistant human laryngeal carcinoma cell line (SQ-20B), tumor cell DNA was transfected into NIH/3T3 cells. A high incidence (six out of six) of raf sequences was found in transfected NIH/3T3 clones and the tumorigenic potential of SQ-20B DNA could be linked to genomic fragments that represent most of the kinase domain of human c-raf-1. An apparently unaltered 3.5-kilobase pair (kb) human c-raf transcript was identified in SQ-20B cells but was not observed in the transfected NIH/3T3 cell clones. Two new transcripts (4.2 kb and 2.6 kb) were found in tumorigenic clones; the large transcript was missing in a very poorly tumorigenic clone. Cytogenetic analysis indicated that the normal autosomes of chromosome 3 were absent in SQ-20B karyotypes and had formed apparently stable marker chromosomes. Unlike the recipient NIH/3T3 cell line, 30 percent of the transformed clone-1 metaphases had minute and double-minute chromosomes representative of amplified DNA sequences. The frequency of the c-raf-1 identification by NIH/3T3 transfection of SQ-20B DNA suggests the presence of some genetic abnormality within this locus.
为了鉴定与耐辐射人喉癌细胞系(SQ - 20B)相关的遗传因素,将肿瘤细胞DNA转染到NIH/3T3细胞中。在转染的NIH/3T3克隆中发现raf序列的高发生率(6个克隆中有6个),并且SQ - 20B DNA的致瘤潜力可能与代表人类c - raf - 1激酶结构域大部分的基因组片段有关。在SQ - 20B细胞中鉴定出一个明显未改变的3.5千碱基对(kb)的人类c - raf转录本,但在转染的NIH/3T3细胞克隆中未观察到。在致瘤克隆中发现了两个新的转录本(4.2 kb和2.6 kb);在一个致瘤性很差的克隆中缺少大的转录本。细胞遗传学分析表明,SQ - 20B核型中不存在3号染色体的正常常染色体,并且形成了明显稳定的标记染色体。与受体NIH/3T3细胞系不同,30%的转化克隆 - 1中期细胞具有代表扩增DNA序列的微小染色体和双微小染色体。通过NIH/3T3转染SQ - 20B DNA鉴定c - raf - 1的频率表明该位点存在一些遗传异常。