煤焦沥青烟提取物致人支气管上皮细胞BEAS-2B染色体不稳定性研究  

Chromosomal Instability of Human Tracheal Epithelial Cells BEAS-2B Induced by Extract of Coal Tar Pitch Fume

作　　者：李智涛[1] 冯艳铭[2] 王威[1] 王丽霞[1] 赵勇[1] 祝寒松[1] 吴卫东[1] 吴逸明[1] 

机构地区：[1]郑州大学公共卫生学院,河南郑州450001 [2]河南科技大学医学院,河南洛阳471003

出　　处：《工业卫生与职业病》2011年第3期129-136,共8页Industrial Health and Occupational Diseases

基　　金：国家自然科学基金(30872095)

摘　　要：目的建立煤焦沥青烟提取物诱导的人支气管上皮细胞BEAS-2B株的恶化模型;观察不同时期细胞的染色体不稳定性与细胞恶性转化之间的关系。方法用四唑蓝(MTT)法测定煤焦沥青烟提取物的细胞毒性,以煤焦沥青烟提取物诱导并观察BEAS-2B细胞传代转化过程中的形态学改变;软琼脂克隆形成实验检测细胞恶性转化能力,流式细胞术检测细胞增殖周期变化,核型分析观察细胞染色体不稳定性。结果煤焦沥青烟提取物的半数致死浓度(LC50)为8.64 mg/L。以诱导剂量(2.0 mg/L)诱导细胞转化,经过30代传代培养,细胞形态发生恶性变化。第30代时,诱导组细胞即能在软琼脂上形成阳性克隆,细胞克隆形成率为21.50‰,明显高于正常对照组和溶剂对照组。流式细胞术检测诱导组G1期细胞比例明显减少,G2/M期细胞比例明显增加。煤焦沥青诱导组从第10代开始细胞二倍体核型的比例明显下降,非整倍体细胞的比例明显增加。随着传代次数增加,染色体不稳定性更明显。细胞克隆形成率和染色体变异的细胞百分比两者呈正相关。结论煤焦沥青烟提取物可以在体外诱导BEAS-2B细胞产生染色体不稳定性并发生恶化。Objective To establish a cell model of malignant transformation with human bronchial epithelial cells BEAS-2B induced by extract of coal tar pitch fume and to observe the relationship between chromosome instability and the cell malignant transformation in different cell generations.Methods MTT assay was used to determine the cytotoxicity of the extract of coal tar pitch fume.After induced by coal tar pitch fume extract at 2.0 mg/L,the morphological changes of BEAS-2B cells were observed in their passage and transformation.In every 10 generations the soft agar cell colony formation rate was used to detect the malignant transformation capability,and traditional methods were used to observe the chromosome karyotype.When morphological changes were shown,cell cycle was detected by flow cytometry.Results Half lethal concentration（LC50）of the fume extract of coal tar pitch was 8.64 mg/L with BEAS-2B cells.BEAS-2B cells were induced at dosage of 2.0 mg/L.After the cells were cultured for 30 passages,the malignant transformation was shown in the induced cells.At the 30th generation,the cell clone formation rate was 21.50‰ in the coal tar pitch group,which was significantly higher than those of the normal control group and DMSO group.The G1 phase cell ratio significantly reduced and G2/M phase cell ratio increased significantly in the coal tar pitch group measured by flow cytometry.Beginning from the 10 th generation,the proportion of diploid cells decreased and aneuploid cells increased distinctively in the coal tar pitch induced group.Chromosomal instability increased as the cell passage increased.Cells in soft agar colony formation rate and the percentage of chromosome aberrations showed a positive correlation.Conclusions The fume extract of coal tar pitch could injury the cell DNA,and could induce BEAS-2B cells to produce chromosomal instability and malignant transformation in vitro.

关 键 词：煤焦沥青烟提取物 细胞恶性转化 染色体不稳定 

分 类 号：R134[医药卫生—劳动卫生]