中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2014年
1期
38-43
,共6页
冯斐斐%张巧%周凡静%吴拥军%吴逸明
馮斐斐%張巧%週凡靜%吳擁軍%吳逸明
풍비비%장교%주범정%오옹군%오일명
肺肿瘤%煤焦沥青烟气提取物%巨噬细胞%肿瘤坏死因子%NF-κB
肺腫瘤%煤焦瀝青煙氣提取物%巨噬細胞%腫瘤壞死因子%NF-κB
폐종류%매초력청연기제취물%거서세포%종류배사인자%NF-κB
Lung neoplasms%Coal tar pitch extract%Macrophages%Tumor necrosis factor%,NF-kappa B
目的 探讨肿瘤坏死因子-α(TNF-α)和NF-κB在人单核巨噬细胞系(THP-1)细胞促煤焦沥青烟气提取物(CTPE)诱导人支气管上皮细胞系(BEAS-2B)细胞恶性转化中的作用.方法 在第10代BEAS-2B和THP-1细胞共培养组中分别加入NF-κB的抑制剂-吡咯烷二硫代氨基甲酸(PDTC)和TNF-α中和抗体,用细胞周期检测、染色体核型分析和软琼脂集落形成实验检测第20代PDTC组和TNF-α中和抗体组BEAS-2B细胞的恶性转化情况,并与第10代、第20代共培养组进行比较;采用实时定量PCR和免疫印迹法(Western blot)法检测各组BEAS-2B细胞中TNFR相关因子2(TRAF2)和细胞周期蛋白D1 (Cyclin D1)基因及其蛋白的表达.结果 第20代PDTC组和INF-α中和抗体组BEAS-2B细胞中S期细胞的比例分别为(33.97％±2.16％)和(34.29％±2.04％),明显低于第20代共培养组(44.46％±0.83％％),差异有统计学意义(P＜0.05).第20代PDTC组和TNF-α中和抗体组100个细胞中出现异常染色体核型的细胞数量为40个和37个,而第20代共培养组为75个,差异有统计学意义(P＜0.05).第20代PDTC组软琼脂集落形成数(15.17±2.48)个,集落形成率为(1.51‰±0.25‰),第20代TNF-α中和抗体组集落形成数为(13.33±2.58)个;集落形成率为(1.33‰±0.26‰),均明显低于20代共培养组f集落形成数:(172.33±12.09)个和集落形成率:(17.23‰±1.20‰)],差异有统计学意义(P＜0.05);PDTC组和TNF-α中和抗体组TRAF2和Cyclin D1基因及其蛋白的表达明显低于第20代共培养组,差异有统计学意义(P＜0.05).结论 TNF-α和NF-κB在THP-1细胞促进煤焦沥青烟气提取物诱导BEAS-2B细胞恶性转化的早期阶段发挥重要的作用,并可能通过影响TRAF2和Cyclin D1的表达促使细胞恶性变发生.
目的 探討腫瘤壞死因子-α(TNF-α)和NF-κB在人單覈巨噬細胞繫(THP-1)細胞促煤焦瀝青煙氣提取物(CTPE)誘導人支氣管上皮細胞繫(BEAS-2B)細胞噁性轉化中的作用.方法 在第10代BEAS-2B和THP-1細胞共培養組中分彆加入NF-κB的抑製劑-吡咯烷二硫代氨基甲痠(PDTC)和TNF-α中和抗體,用細胞週期檢測、染色體覈型分析和軟瓊脂集落形成實驗檢測第20代PDTC組和TNF-α中和抗體組BEAS-2B細胞的噁性轉化情況,併與第10代、第20代共培養組進行比較;採用實時定量PCR和免疫印跡法(Western blot)法檢測各組BEAS-2B細胞中TNFR相關因子2(TRAF2)和細胞週期蛋白D1 (Cyclin D1)基因及其蛋白的錶達.結果 第20代PDTC組和INF-α中和抗體組BEAS-2B細胞中S期細胞的比例分彆為(33.97％±2.16％)和(34.29％±2.04％),明顯低于第20代共培養組(44.46％±0.83％％),差異有統計學意義(P＜0.05).第20代PDTC組和TNF-α中和抗體組100箇細胞中齣現異常染色體覈型的細胞數量為40箇和37箇,而第20代共培養組為75箇,差異有統計學意義(P＜0.05).第20代PDTC組軟瓊脂集落形成數(15.17±2.48)箇,集落形成率為(1.51‰±0.25‰),第20代TNF-α中和抗體組集落形成數為(13.33±2.58)箇;集落形成率為(1.33‰±0.26‰),均明顯低于20代共培養組f集落形成數:(172.33±12.09)箇和集落形成率:(17.23‰±1.20‰)],差異有統計學意義(P＜0.05);PDTC組和TNF-α中和抗體組TRAF2和Cyclin D1基因及其蛋白的錶達明顯低于第20代共培養組,差異有統計學意義(P＜0.05).結論 TNF-α和NF-κB在THP-1細胞促進煤焦瀝青煙氣提取物誘導BEAS-2B細胞噁性轉化的早期階段髮揮重要的作用,併可能通過影響TRAF2和Cyclin D1的錶達促使細胞噁性變髮生.
목적 탐토종류배사인자-α(TNF-α)화NF-κB재인단핵거서세포계(THP-1)세포촉매초력청연기제취물(CTPE)유도인지기관상피세포계(BEAS-2B)세포악성전화중적작용.방법 재제10대BEAS-2B화THP-1세포공배양조중분별가입NF-κB적억제제-필각완이류대안기갑산(PDTC)화TNF-α중화항체,용세포주기검측、염색체핵형분석화연경지집락형성실험검측제20대PDTC조화TNF-α중화항체조BEAS-2B세포적악성전화정황,병여제10대、제20대공배양조진행비교;채용실시정량PCR화면역인적법(Western blot)법검측각조BEAS-2B세포중TNFR상관인자2(TRAF2)화세포주기단백D1 (Cyclin D1)기인급기단백적표체.결과 제20대PDTC조화INF-α중화항체조BEAS-2B세포중S기세포적비례분별위(33.97％±2.16％)화(34.29％±2.04％),명현저우제20대공배양조(44.46％±0.83％％),차이유통계학의의(P＜0.05).제20대PDTC조화TNF-α중화항체조100개세포중출현이상염색체핵형적세포수량위40개화37개,이제20대공배양조위75개,차이유통계학의의(P＜0.05).제20대PDTC조연경지집락형성수(15.17±2.48)개,집락형성솔위(1.51‰±0.25‰),제20대TNF-α중화항체조집락형성수위(13.33±2.58)개;집락형성솔위(1.33‰±0.26‰),균명현저우20대공배양조f집락형성수:(172.33±12.09)개화집락형성솔:(17.23‰±1.20‰)],차이유통계학의의(P＜0.05);PDTC조화TNF-α중화항체조TRAF2화Cyclin D1기인급기단백적표체명현저우제20대공배양조,차이유통계학의의(P＜0.05).결론 TNF-α화NF-κB재THP-1세포촉진매초력청연기제취물유도BEAS-2B세포악성전화적조기계단발휘중요적작용,병가능통과영향TRAF2화Cyclin D1적표체촉사세포악성변발생.
Objective To characterize the role of tumor necrosis factro-o (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 tumorigenecity,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-αt 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-αt 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 D 1.
