肿瘤坏死因子-α和核因子-κB在巨噬细胞样THP-1细胞促进煤焦油沥青提取物诱导的人支气管上皮细胞致瘤性转化中发挥作用。
[Tumor necrosis factor-α and NF-κB play a role in macrophage-like THP-1 cells promoting coal tar pitch extract-induced tumorigenic transformation of human bronchial epithelial cells].
作者信息
Feng Feifei, Zhang Qiao, Zhou Fanjing, Wu Yongjun, Wu Yiming
机构信息
Department of Toxicology, College of Public Health, Zhengzhou University, Zhengzhou 450001, China.
Department of Toxicology, College of Public Health, Zhengzhou University, Zhengzhou 450001, China. E-mail:
出版信息
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 2014 Jan;32(1):38-43.
OBJECTIVE
To characterize the role of tumor necrosis factor-α (TNF-α) and NF-κB play a role in macrophage-like THP-1 cells promoting coal tar pitch extract (CTPE)-induced tumorigenic transformation of human bronchial epithelial cells (BEAS-2B).
METHODS
From passage 10, CTPE-induced BEAS-2B cells cocultured with THP-1 cells were treated with NF-κB inhibitor-Pyrrolidine dithiocarbamate (PDTC) every 3 passages and TNF-α antibody every passage. Alterations of cell cycle, karyotype and colony formation in soft agar of BEAS-2B cells at passages 20, indicative of tumorigenicity, were determined, respectively. In addition, mRNA and protein levels of TNF receptor associated factor2 (TRAF2) and Cyclin D1 in BEAS-2B cells were measured with Real Time-PCR and Western blot, respectively.
RESULTS
The percentages of S-phase BEAS-2B cells at passage 20 in PDTC group and TNF-α antibody group were (33.97±2.16)% and (34.29±2.04)% respectively, which were less than that in Co-culture+CTPE group of 20th passage [(44.46±0.83)%], P < 0.05; The number of cells with aneuploidy in 100 cells in 20th passage PDTC group and TNF-α antibody group were 40 and 37, and there were significantly different when comparing to that of 20th passage Co-culture+CTPE group (75); The number of colony formation and the rate of colony formation of BEAS-2B cells in soft agar at passage 20 in PDTC group were (15.17±2.48) and (1.51‰±0.25‰), (13.33±2.58)and (1.33‰±0.26‰) in TNF-α antibody group, which were less that those in 20th passage Co-culture+CTPE group [(172.33±12.09) and (17.23‰±1.20‰)], P < 0.05; at the same time, the mRNA and protein levels of TRAF2 and Cyclin D1 in BEAS-2B cells were decreased after PDTC and TNF-α antibody treatment.
CONCLUSION
TNF-α and NF-κB could play an important role in THP-1 cells promoting coal tar pitch extract-induced tumorigenic transformation of BEAS-2B cells by influencing the expression of TRAF2 and Cyclin D1.
目的
探讨肿瘤坏死因子-α（TNF-α）和核因子-κB（NF-κB）在巨噬细胞样THP-1细胞促进煤焦油沥青提取物（CTPE）诱导人支气管上皮细胞（BEAS-2B）致瘤性转化中的作用。
方法
从第10代开始，每3代用NF-κB抑制剂-吡咯烷二硫代氨基甲酸盐（PDTC）处理与THP-1细胞共培养的CTPE诱导的BEAS-2B细胞，每代用TNF-α抗体处理。分别测定第20代BEAS-2B细胞的细胞周期、核型变化及软琼脂集落形成情况，这些指标可指示致瘤性。此外，分别用实时定量PCR和蛋白质免疫印迹法检测BEAS-2B细胞中肿瘤坏死因子受体相关因子2（TRAF2）和细胞周期蛋白D1的mRNA和蛋白水平。
结果
第20代PDTC组和TNF-α抗体组BEAS-2B细胞S期百分比分别为（33.97±2.16）%和（34.29±2.04）%，低于第20代共培养+CTPE组[（44.46±0.83）%]，P<0.05；第20代PDTC组和TNF-α抗体组100个细胞中出现非整倍体的细胞数分别为40和37，与第20代共培养+CTPE组（75）相比有显著差异；第20代PDTC组BEAS-2B细胞在软琼脂中的集落形成数和集落形成率分别为（15.17±2.48）和（1.51‰±0.25‰），TNF-α抗体组为（13.33±2.58）和（1.33‰±0.26‰），均低于第20代共培养+CTPE组[（172.33±12.09）和（17.
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