In vitro transformation of rat esophageal epithelial cells with N-nitrosobenzylmethylamine.
In vitro transformation of rat esophageal epithelial cells with N-nitrosobenzylmethylamine.
复制标题
N-亚硝基苄基甲胺体外转化大鼠食管上皮细胞。
DOI：
10.1093/carcin/3.6.629
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发表时间：
1982
期刊：
影响因子：
4.7
通讯作者：
Knipe,SM
中科院分区：
文献类型：
--
作者：
Stoner,GD;Babcock,MS;Cothern,GA;Klaunig,JE;Gunning3rd,WT;Knipe,SM
采用组织块/细胞培养系统，在体外用N-亚硝基-N-苄基-N-甲胺（BMNA）处理大鼠食管上皮细胞。将每组12个食管外植体培养物暴露于0.0、2.5、5.0或10.0 μg BMNA/ml培养基两次（第1天和第7天）。孵育60-90天后，用所有三种浓度的BMNA处理的原代培养物中的上皮细胞可以传代培养，并形成细胞系。原代培养物的数量和随后发育的上皮细胞系的数量是致癌剂剂量依赖性的。细胞系只能从致癌物处理的外植体中建立。电子显微镜检查显示BMNA处理的细胞系含有食管上皮细胞的形态学标记;即，许多张力细丝和连接复合体，即使经过长期的继代培养。用免疫荧光法检测，细胞与制备的小鼠皮肤前角蛋白（K1和K2）抗体呈阳性反应。两种细胞系（来自5 μg BMNA/ml组）能够在软琼脂中生长，并在注射到同基因受体中时产生可触知的肿瘤。这些肿瘤具有鳞状细胞癌的组织学特征。
Using an explant/cell culture system, rat esophageal epithelial cells were transformedin vitroby exposure to N-nitroso-N-benzyl-N-methylamine (BMNA). Twelve esophageal explant cultures per group were exposed twice (at days 1 and 7) to 0.0, 2.5, 5.0 or 10.0 μg BMNA/ml of medium. After incubation for 60–90 days, epithelial cells in primary cultures treated with all three concentrations of BMNA could be subcultured and cell lines were developed. The number of primary cultures and the number of subsequently developed epithelial cell lines was carcinogen-dose-dependent. Cell lines could only be established from carcinogen treated explants. Electron microscopy revealed that the BMNA-treated cell lines contained morphological markers of esophageal epithelial cells; i.e., numerous tonofilaments and junctional complexes, even after prolonged subculture. By immunofluorescence, the cells reacted positively with antibodies prepared to mouse skin prekeratins (K1and K2). Two cell lines (from the 5 μg BMNA/ml group) were able to grow in soft agar and produce palpable tumors upon injection into syngeneic recipients. These tumors possessed the histological features of squamous cell carcinomas.