烟草中甲基亚硝胺吡啶基丁酮诱发人支气管上皮细胞恶性转化的研究

Study on malignant transformation of human bronchial epithelial cells induced by tobacco-specific nitrosamine NNK

目的研究吸烟致肺癌过程中人类支气管上皮细胞恶变机制,探索一种较快捷的体外研究吸烟致肺癌的途径。方法采用细胞毒性实验确定亚硝胺吡啶基丁酮(NNK)水溶液转化剂量为600μg/ml,应用NNK多次染毒法对人支气管上皮细胞系(16HBE)进行转化,对转化组织和细胞分别进行病理学检查和透射电镜与扫描电镜分析。结果16HBE细胞染毒至第23代,细胞呈恶性形态;可以在软琼脂上形成集落(锚着非依赖性实验为阳性);裸鼠成瘤实验为阳性。对裸鼠成瘤组织进行病理组织学检查,证实为低分化鳞状细胞癌,并取裸鼠成瘤组织原代培养细胞作扫描电镜和透射电镜检查,结果显示为瘤组织培养细胞具有恶性型肿瘤细胞的特征。结论NNK致16HBE细胞恶性转化能力较强,为深入研究人类支气管上皮细胞恶变机制提供了理想的生物模型。

Objective To explore the mechanism of the malignant transformation of human bronchial epithelial cell （16HBE） during the lung cancer induced by smoking, and find a shortcut methods for the study of lung cancer induced by smoking. Methods We con_firmed the transform dosage was 600 μg/ml for 4-（ N-methyl-N-nitrosamino）-1-（3-pyridly）-1-butanone （NNK） solution by cell toxicity test. 16HBE cells treated with NNK setp by step, and the transformed cell and tissue were analysed by morphology and electron misroscope melthods. Results The 23th passage cells were introduced by NNK, colony formation efficiency of transformed cells in semisold ager showed significant increase（ P 〈 0.01 ） and dose-dependent effects and could grow into tumor in nude mice. The tumor was poor differentiated squamous cell carcinoma con-firmed ,by histopathological examination. Conclusion NNK has a strongly transformed ability.This culture system provides a potential tool for the study of cell and molecular mechanism in the carcinogenesis of human bronchial epithelial cells.