Manoharan T H, Burgess J A, Ho D, Newell C L, Fahl W E
Carcinogenesis. 1985 Sep;6(9):1295-301. doi: 10.1093/carcin/6.9.1295.
C3H/10T1/2-CL8 mouse cells were shown to take up and express a plasmid-cloned drug resistance gene (Ecogpt) after DNA transfection at a frequency (2-6 X 10(-4) which is acceptable for routine recovery of gene-transformed populations. Transfection of 10T1/2 cells with a mutant c-Ha-ras oncogene (pEJ6.6 plasmid) results in neoplastically transformed 10T1/2 cell populations as judged by colony morphology and tumorigenic growth in nude mice. The levels of mutant c-Ha-ras gene integration and expression in the tumorigenic cell populations and 10T1/2 cell controls were determined, and the highest level of mutant ras transcript was seen in the most tumorigenic cell population. A preliminary comparison of 10T1/2 and NIH/3T3 cells showed similar frequencies for pEJ 6.6-induced transformed foci and a similar lack of sensitivity to the transforming effects of a cloned B-lym oncogene. The results identify a genetic event, which has previously been shown to be carcinogen-inducible, that is permissive for neoplastic transformation of the widely used carcinogen-transformable 10T1/2 mouse cell line.
已证明C3H/10T1/2 - CL8小鼠细胞在DNA转染后能够摄取并表达质粒克隆的耐药基因（Ecogpt），其频率为（2 - 6×10⁻⁴），这对于常规回收基因转化群体而言是可以接受的。用突变型c - Ha - ras癌基因（pEJ6.6质粒）转染10T1/2细胞，根据集落形态和在裸鼠中的致瘤生长情况判断，会产生肿瘤转化的10T1/2细胞群体。测定了致瘤细胞群体和10T1/2细胞对照中突变型c - Ha - ras基因整合和表达的水平，在致瘤性最强的细胞群体中观察到了最高水平的突变型ras转录本。对10T1/2细胞和NIH/3T3细胞的初步比较显示，pEJ 6.6诱导的转化灶频率相似，并且对克隆的B - lym癌基因的转化作用同样缺乏敏感性。这些结果确定了一个遗传事件，该事件先前已被证明可被致癌物诱导，并且对于广泛使用的可被致癌物转化的10T1/2小鼠细胞系的肿瘤转化是允许的。
