Neoplastic transformation of primary tracheal epithelial cell cultures.
Neoplastic transformation of primary tracheal epithelial cell cultures.
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原代气管上皮细胞培养物的肿瘤转化。
DOI：
10.1093/carcin/4.4.369
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发表时间：
1983
期刊：
影响因子：
4.7
通讯作者：
P. Nettesheim
中科院分区：
文献类型：
--
作者：
S.Balakrishna Pai;Vernon E. Steele;P. Nettesheim
用化学致癌剂N-甲基-N '-硝基-N-亚硝基胍（MNNG）处理原代培养的大鼠气管上皮细胞，以定量研究肿瘤转化的早期事件。从无特定病原体的Fischer-344大鼠的气管中分离上皮细胞，并将其铺在胶原包被的组织培养皿上。为了测定细胞毒性，在第1天将细胞暴露于各种浓度的MNNG 3小时，并在第7天测定集落形成效率（CFE）。与对照培养物相比，浓度为0.1微克/毫升的MNNG没有降低CFE，而1微克/毫升的MNNG使CEF降低了75%。对于转化研究，原代细胞培养物在第1天和第17天之间接受单次暴露于MNNG（0.1-0.6 μ g/ml）或多次暴露于0.1 μ g/ml MNNG 3 h。在致癌物暴露的文化，形态学改变的病灶出现在第18天，可识别的高细胞密度。根据MNNG浓度，观察到1和8%之间的转化频率。当对照培养物停止增殖并从培养皿脱落时，含有改变的病灶的培养物在第三和第四周期间继续生长。超过40%的多次暴露于MNNG的培养物获得细胞系状态，并且可以传代培养大于或等于20次。30个对照培养物中没有一个成为细胞系。70%的MNNG暴露的细胞系显示锚定独立的生长表型在第20代琼脂糖生长判断。10个培养物中有4个暴露于6或8次MNNG，接种到裸鼠后在第20代形成浸润性鳞状细胞癌。基于这些和以前的研究，我们认为，不受限制的细胞复制是一个早期的关键事件，在致癌物暴露的上皮细胞群，肿瘤转化之前。
Primary cultures of rat tracheal epithelial cells were treated with the chemical carcinogen N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) to quantitatively study the early events during neoplastic transformation. Epithelial cells were dissociated from tracheas of specific-pathogen-free Fischer-344 rats and were plated on collagen-coated tissue culture dishes. To determine cytotoxicity, cells were exposed on day 1 to various concentrations of MNNG for 3 h and colony forming efficiency (CFE) was determined on day 7. MNNG at a concentration of 0.1 microgram/ml did not decrease CFE as compared to the control cultures, whereas 1 microgram/ml reduced the CEF by 75%. For transformation studies, primary cell cultures received single exposures to MNNG (0.1-0.6 microgram/ml) or multiple exposures to 0.1 microgram/ml of MNNG for 3 h between days 1 and 17. In carcinogen-exposed cultures, morphologically altered foci appeared on day 18, recognizable by high cell density. Transformation frequencies between 1 and 8% were observed depending on MNNG concentration. Cultures containing altered foci continued to grow during the third and fourth week when control cultures had ceased to proliferate and exfoliated from the dish. Over 40% of the cultures which received multiple exposures to MNNG acquired cell line status and could be subcultured greater than or equal to 20 times. None of the 30 control cultures became cell lines. Seventy per cent of MNNG-exposed cell lines showed the anchorage independent growth phenotype at passage 20 as judged by growth in agarose. Four of 10 cultures exposed either 6 or 8 times to MNNG formed invasive squamous cell carcinomas at passage 20 upon inoculation into nude mice. Based on these and previous studies, we feel that unrestricted cell replication is an early key event in carcinogen-exposed epithelial cell populations, preceding neoplastic transformation.