TRAF1在霍奇金淋巴瘤细胞中的表达和作用机制的探讨  

Expression and function of TRAF1 in Hodgkin＇ s lymphoma cells

作　　者：王文娟[1] 国风[2] 孙爱宁[1] 周鹏[2] 马亮[2] 

机构地区：[1]苏州大学附属第一医院血液科、江苏省血液研究所,215006 [2]苏州大学附属第一医院中心实验室

出　　处：《中华血液学杂志》2010年第1期29-33,共5页Chinese Journal of Hematology

基　　金：江苏省卫生厅面上项目（H200877）;江苏省六大人才高峰资助项目

摘　　要：目的探讨肿瘤坏死因子受体相关因子（TRAF）1以及CD30-TRAF1信号通路在B细胞性霍奇金淋巴瘤细胞中的作用。方法分别采用荧光定量RT—PCR和Westernblot法检测B细胞性霍奇金淋巴瘤细胞株TRAF1mRNA和蛋白质表达水平，并进一步研究CD30信号活化对TRAF1表达的作用。采用RNA干扰技术研究TRAF1沉默对霍奇金淋巴瘤细胞存活的影响，并通过TransAM方法定量分析核转录因子（NF）-κB的DNA结合活性的变化。通过Westernblot法测定NAPK／ERK信号通路中JunB蛋白的表达。结果B细胞性霍奇金淋巴瘤细胞株IA28和KM—H2细胞中TRAF1在mRNA和蛋白质水平均表达。CD30配体作用后IA28细胞中TRAF1mRNA相对表达水平自3．50±0．20上调至7．26±0．23；蛋白相对表达水平自2．31±0．35上升至4．53±0．55。TRAF1沉默后凋亡细胞百分率由（5．7±1．2）％上升至（27．7±5．8）％，并伴随p50和RelA活力的下降。JunB的表达在TRAF1沉默细胞中未发生明显变化。结论TRAF1在B细胞系来源的霍奇金淋巴瘤细胞中表达增加，并受到CD30信号通路活化的调控。TRAF1为介导经典性NF—κB活性的关键性调节分子，参与霍奇金淋巴瘤细胞的抗凋亡活性。Objective To investigate the function of tumor necrosis factor receptor-associated factor 1 （ TRAF1 ） and CD30-TRAF1 signaling in Hodgkin＇ s lymphoma. Methods Endogenous and CD30 ligandinduced TRAF1 expression at mRNA and protein levels were examined by quantitative RT-PCR and Western blot analyses, respectively. RNA interference was performed to silence the expression of TRAF1 in IA28 cells and examine its effect on cell survival. ELISA-based NF-κB family transcription factor activity assay was performed to quantify the KB DNA-binding activity in nuclear extracts. The expression of JunB was measured by Western blot. Results TRAFI expression was detected at both mRNA and protein levels in B cell-derived lymphoma cell lines （IA28 and KM-H2）. CD30 activation via binding to CD30 ligand induced the TRAF1 expression, the relative mRNA expression was increased to 7.26 ± 0.23 from 3.50 ± 0.20, the relative protein expression was increased to 4.53 ± 0.55 from 2.31± 0.35. The apoptosis rate was increased to （27.7 ±5.8 ） % in TRAFl-silenced IA28 cells compared to （5.7 ±1.2） % in control cells. The p50 and RelA DNA- binding activity were decreased in TRAF1-silenced IA28 ceils. The expression of JunB upon CD30 ligand stimulation was not changed in TRAFl-silenced IA28 cells. Conclusions TRAF1 is overexpressed in B cell- derived Hodgkin＇ s lymphoma cells, which is regulated by CD30 signaling pathway. TRAFl is a crucial molecule mediating the activation of the classical NF-κB activity, which further facilitates the anti-apoptosis.

关 键 词：TNF受体相关因子1 霍奇金病 配体 CD30 NF—κB 

分 类 号：R733.1[医药卫生—肿瘤] R733.7[医药卫生—临床医学]