单核巨噬细胞在煤焦沥青烟提取物诱导永生化人支气管上皮细胞恶变中的作用

The Effects of monocyte-macrophages on malignant transformation of human bronchial epithelial cells induced by extracts from coal tar pitch

目的探讨单核巨噬细胞（THP-1）在煤焦沥青烟提取物（coaltarpitch，CTP）致永生化人支气管上皮细胞humanbronchialepithelialcells，BEAS-2B）恶变过程中的作用及肿瘤坏死因子0【（tumornecrosisfactoralpha，TNF-α）在该过程中的表达。方法以THP．1和BEAS．2B为研究对象，设立CTP组、苯并（a）芘[B（a）P]组（阳性对照组）、二甲亚砜对照组（溶剂对照组）、BEAS-2B与THP-1共培养组，建立细胞恶性变模型。应用软琼脂集落形成实验、染色体数目畸变分析、流式细胞仪测定细胞周期中不同培养时期（10、20、30代）细胞的恶变情况，利用ELISA方法测定CTP组及共培养组细胞培养上清中TNF-α的含量。结果染色体数目异常在实验的早期（10代）已经显现，表现为非整倍体和多倍体比例增加，二倍体数目减少。在第20代：共培养组克隆形成率（17．63‰±0．97‰）明显高于CTP组（13．94‰±0．84‰）和阳性对照组组（12．96‰±1．62‰），差异有统计学意义（P〈0．05）；共培养组S期细胞比例（44．49％±0．68％）明显高于CTP组（38．19％±1．26％）和阳性对照组（36．41％±1．19％），差异有统计学意义（P〈0．05）。共培养组细胞培养上清中TNF-α含量明显高于CTP组，差异有统计学意义（P〈0．01）。结论THP-1能够加速CTP诱导的BEAS．2B恶变，增加TNF-α的表达水平。

Objective To study the effects of monocyte-macrophages （THP-1） in malignant transfor- mation of human bronchial epithelial cells （BEAS-2B） cells induced by coal tar pitch （CTP） and the expres- sion of TNF-α in the process of the cell malignant transformation. Methods BEAS-2B cells and THP-1 Cells were divided into four groups： coal tar pitch （CTP） group, benzo （a）pyrene [B（a）P] group, dimethyl sulfoxide （DMSO） group, BEAS-2B and THP-1 co-culture （co-culture group） group. Carcinogenesis model was estab- lished. The soft agar colony formation, chromosome aberrations and cell cycle tests were used to detect the cel- lular malignant transformation. The ELISA assay was utilized to measure the levels of TNF-α in the supernatant of CTP group and co-culture group. Results The chromosome number abnormalities could be observed in early stage of the experiment （the 10th generation cells）, which showed the increased ratio of aneuploid to polyploid, and the decreased number of diploid. The colony formation rate of co-culture group （the 20th generation cells） was 17.63‰±0.97‰, which was significantly higher than that （13.94‰±0.84‰） of CTP group and that （ 12.96‰±1.62‰） of B （a）P group （P〈0.05）. The proportion of S phase cells in the co-culture group was 44.49%±0.68%, which was significantly higher than that （38.19%±1.26%） of CTP group and that （36.41%±1.19%） of B （a）P group （P〈0.05）. The TNF-α level in the co-culture group was significantly higher than that in CTP group （P=0.001）. Conclusion Monocyte-Macrophages can accelerate the malignant transformation of BEAS- 2B cells induced by CTP and increase the expression level of TNF-α.