烹调油烟冷凝物诱导永生化人支气管上皮细胞恶性转化的研究

Study for Malignant Transformation of Immortalized Human Bronchial Epithelial(BEP-2D) Cells Induced by Cooking Oil Fumes Condensation

目的用烹调油烟冷凝物(COFC)诱导正常永生化人支气管上皮细胞(BEP-2D)慢性恶性转化,构建恶性转化的细胞模型,以探讨COFC的危害。方法通过MTT法确定诱导物的染毒剂量,以COFC处理细胞,用血清抗性实验和锚着独立生长实验鉴定细胞恶性转化的倾向和恶性特征,比较对照组和实验组抗血清促分化的能力和集落形成率,并将细胞接种于裸鼠,观察其成瘤情况。结果以50μg/mL的COFC对细胞进行染毒,传至第20代细胞,与对照组相比,其抗血清促分化的能力差异均有统计学意义(P<0.01);传至第35代,转化细胞的锚着独立生长实验克隆形成率差异均有统计学意义(P<0.05),并接种裸鼠成瘤,病理检查结果为低分化鳞癌。结论 COFC可诱导正常永生化人支气管上皮细胞发生恶性转化,提示烹调油烟对人体具有致癌致畸作用。

Objective To find a good method for chronic malignant tranformation of immortalized human bronchial epithelial（BEP-2D） cells induced by cooking oil fumes condensation（COFC）,establish a model of malignant tranformation of BEP-2D cells and discuss the harm of COFC.Methods Cells was divided into control group and experimental group.MTT was used to decide the chronic exposure doses of the COFC.The malignant tendency of transformed BEP-2D cells was identified by anti-blood serum experiment and the malignant features of transformed BEP-2D cells were identified by the method of semisolid agar culture.The capability of COFC inducing differentiation in anti-blood serum experiment and the colony forming frequency of BEP-2D cells in semisolid agar medium in each group were compared.Results The best method for malignant tranformation of BEP-2D cells induced by the COFC was that the BEP-2D cells were treated with COFC at doses of 50 μg/mL respectively.Inoculated to the 20 generations,the cells inoculative efficiency were obviously disparity in the comparison of control group（P 0.01）.Inoculated to the 35th generation,the anchorage independent growth was appeared in the P35（P 0.05）.The P35 BEP-2DCOFC cells formed tumors in nude mice,and tumors were poorly differentiated to squamous cell carcinoma confirmed by histopathological examination.Conclusion The malignant tranformation of immortalized human bronchial epithelial（BEP-2D） cells could be successfully induced by the COFC at the dose of 50 μg/mL,and the prompt cooking lampblack had the carcinogenicity to the human body.