Haas M, Isakov J, Howell S B
Department of Biology, University of California, San Diego, La Jolla 92093.
Mol Biol Med. 1987 Oct;4(5):265-75.
Activation of ras genes in human ovarian carcinomas and ovarian carcinoma cell lines was tested by transfection of NIH 3T3 cells with high molecular weight DNA extracted from fresh tumors or from cell lines. Of 18 ovarian tumors and tumor cell lines that were exhaustively tested, only one yielded DNA active in focus-induction in this assay. Southern blot analysis of DNA in serially transformed NIH 3T3 foci revealed restriction fragments that hybridized with a probe specific for the human N-ras oncogene and with human repetitive (Alu) sequences. Individually transformed foci contained N-ras hybridizing fragments of different sizes. Northern blots of RNA extracted from different transformants revealed the expression of different transcripts hybridizing with the N-ras probe. The transformants also expressed a 700 base RNA that hybridized with a human B-lym probe. The low prevalence of ras activation in ovarian tumors as measured by transforming capability suggests that ras activation does not play an important role in the development of most human ovarian tumors.
通过用从新鲜肿瘤或细胞系中提取的高分子量DNA转染NIH 3T3细胞，检测人卵巢癌及卵巢癌细胞系中ras基因的激活情况。在对18个卵巢肿瘤和肿瘤细胞系进行全面检测后，在此检测中只有一个产生了具有焦点诱导活性的DNA。对连续转化的NIH 3T3焦点中的DNA进行Southern印迹分析，发现了与人类N-ras癌基因特异性探针以及人类重复（Alu）序列杂交的限制性片段。单个转化焦点包含不同大小的与N-ras杂交的片段。从不同转化体中提取的RNA进行Northern印迹分析，发现了与N-ras探针杂交的不同转录本的表达。这些转化体还表达了一种与人类B-lym探针杂交的700碱基RNA。通过转化能力测量，卵巢肿瘤中ras激活的低发生率表明ras激活在大多数人类卵巢肿瘤的发生中不发挥重要作用。
