香烟烟雾诱发BEAS-2B细胞体外恶性转化的研究  

Study on malignant transformation of immortalized human bronchial epithelial cells BEAS-2B induced by cigarette smoke in vitro

作　　者：杜厚兵[1] 童建[1] 聂继华[1] 张素萍[1] 吴昭昭[1] 刘海云[1] 李建祥[1] 

机构地区：[1]苏州大学医学部放射医学与公共卫生学院卫生毒理学教研室江苏省放射医学与防护重点实验室,江苏苏州215123

出　　处：《毒理学杂志》2011年第1期17-21,共5页Journal of Toxicology

基　　金：国家自然科学基金项目(30671784);苏州大学医学部研究生创新基金

摘　　要：目的通过香烟烟雾体外染毒诱发细胞恶性转化,建立吸烟致肺癌的细胞模型。方法确定合适染烟浓度及时间后,将永生化人支气管上皮细胞系BEAS-2B细胞分为对照组(C)和20%烟雾浓度组(S)。以每孔1×105细胞接种于Transwell培养皿,贴壁后置于气体染毒装置中,香烟烟雾持续泵入直接对细胞染毒。检测染毒后5、10、15、20代细胞恶性转化情况,流式细胞仪分析细胞周期和凋亡变化。结果 BEAS-2B细胞在20%烟雾浓度下染毒5代后,出现生长动力学和形态学改变,生长失去接触抑制,获得对血清诱导分化的抗性和锚着独立性生长能力。G1期细胞百分比明显降低,S期细胞百分比明显升高,G2期细胞百分比先升高后降低,各代细胞凋亡率均明显降低。结论香烟烟雾染毒能够诱发BEAS-2B细胞体外转化,且随染毒代数的增加呈现剂量效应关系,可作为体外研究吸烟致肺癌的细胞模型。Objective To establish a model of malignant transformation of human cells in vitro to study lung cancer induced by cigarette smoke.Methods When the time and concentration had been determined,the immortalized human bronchial epithelial cells BEAS-2B were divided into a control group（C） and a concentration 20% cigarette smoke group（S）.1 × 105 cells were planted onto Transwell membrane one day before exposure to cigarette smoke with a pump in a gas inhalation box.After the exposure cells were trypsinized into dishes for further growth and phenotype transformation was detected in order to compare the results due to cigarette smoke exposure.Cell cycle and apoptotic rate were detected through Flow Cytometer.Results There are series of sequential steps emceed among the transformed cells from the fifth generation,including altered growth kinetics and morphology,loss of contact inhibition resistance to serum and obtain anchorage independent growth ability.The percentage of G1 phase in the fifth generation cells was significantly lower,and that of the S phase was significantly higher with the G2 phase percentage increasing first and then decreasing.The apoptotic rates were significantly lower in all the cell generations.Conclusion BEAS-2B cells could be malignantly transformed by smoke exposure in vitro,with the increase of exposure there has a dose-effect relation,which could be used as a cell model in lung bronchial carcinogenesis.

关 键 词：香烟烟雾 体外染毒 BEAS-2B细胞 恶性转化 

分 类 号：R99[医药卫生—毒理学]