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| 低剂量烟草悬凝物诱导正常永生化人支气管上皮细胞慢性恶性转化 |
| 张诚1,吴庆琛1*,李强1,朱茂祥2,杨陟华2,潘秀颉2,谢俊豪1 |
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| (1.重庆医科大学附属第一医院胸心外科,重庆 400016;2.军事医学科学院放射与辐射医学研究所,北京 100850) |
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| 摘要: |
| 目的：用低剂量烟草悬凝物模拟吸烟环境诱导正常永生化人支气管上皮细胞慢性恶性转化，构建恶性转化的细胞模型。方法：通过MTT法及细胞亚急性毒性存活能力检测实验确定慢性诱导物的剂量，以不同剂量的烟草悬凝物对细胞进行单次或多次染毒,以不染毒细胞为对照。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，比较各组抗血清促分化的能力和集落形成率。结果：以体积分数为0.5、1、2 μl/ml的烟草悬凝物对细胞进行单次和多次染毒，各相应剂量组多次染毒和单次染毒与对照组相比，传至第25代细胞，其抗血清促分化的能力均差异明显（P<0.05）；传至第38代，成功建立细胞恶性转化模型，细胞复层生长，无接触性抑制，染色体异常，转化细胞的锚着独立生长实验克隆形成率均明显高于对照组（P<0.05）。多次染毒组的恶性转化趋势比单次染毒组强，而且剂量反应关系呈现良好的直线相关（r=0.969,y=42x）。结论：用0.5～2 μl/ml烟草悬凝物诱导的正常永生化人支气管上皮细胞恶性转化，可作为模拟吸烟环境下细胞慢性恶性转化的理想模型。 |
| 关键词: 烟草悬凝物 致癌性试验 人支气管上皮细胞 慢性恶性转化 |
| DOI：10.3724/SP.J.1008.2008.00150 |
| 投稿时间：2007-10-26修订日期：2007-11-26 |
| 基金项目: |
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| Low dose of cigarette smoke condensate induces chronic malignant transformation of immortalized human bronchial epithelial cells |
| ZHANG Cheng1,WU Qing-chen1*,LI Qiang1,ZHU Mao-xiang2,YANG Zhi-hua2,PAN Xiu-jie2,XIE Jun-hao1 |
| (1.Department of Cardiothoracic Surgery，The First Affiliated Hospital of Chongqing Medical University，Chongqing 400016, China;2.Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850) |
| Abstract: |
| Objective：To establish a chronic malignant transformation model of immortalized human bronchial epithelial cell line BEP2D by low dose cigarette smoke condensate (simulating smoking environment).Methods: The chronic dose of cigarette smoke condensate was determined by MTT assay and the colony formation test of BEP2D cells. BEP2D cells were exposed to cigarette smoke condensate once or for multiple times; unexposed cells were taken as control. The malignant tendency of BEP2D cells was identified by antiserum experiment and the malignant features of transformed BEP2D cells were identified by semisolid agar culture. The differentiation ability of BEP2D cells in antiserum experiment and the colony forming rates of BEP2D cells were compared between different groups. Results: The BEP2D cells were exposed to 0.5,1 and 2 μl/ml of cigarette smoke condensate once or for multiple times and were cultured for 25 generations;the differentiation abilities of BEP2D cells(the 25th generation) was significantly different between the cigarette smoke condensate exposed groups (at 0.5,1 and 2 μl/ml) and the normal control group (P<0.05). The cell malignant transformation model was successfully established in the cells of the 38th generation; the cells had multilayer growth and had no contacting inhibition, with chromosome abnormality. The colony forming rates in the semisolid agar culture test was significantly higher in all smoke condensate exposed groups than in control group(P<0.05). The doseresponse relationship showed a good linear correlation (r=0.969, y=42x, P<0.05). Conclusion: The malignant transformation of immortalized human bronchial epithelial cells can be successfully induced by cigarette smoke condensate at 0.52 μl/ml, which offers an ideal model for simulating smoking environment induced chronic malignant transformation. |
| Key words: cigarette smoke condensate carcinogenicity tests chronic malignant transformation bronchial epithelial cells |
