EB病毒LMP1在鼻咽癌细胞系中通过NF-κB、AP-1促进IL-8分泌  

IL-8 production in nasopharyngeal carcinoma cell line via an NF-κB or AP-1 signaling pathway mediated by Epstein-Barr virus-encoded LMP1

作　　者：王承兴[1] 顾焕华[2] 邓锡云[2] 李晓艳[2] 夏林庆[2] 易薇[2] 翁新宪[2] 曹亚[2] 

机构地区：[1]中南大学湘雅医院口腔颌面外科 [2]中南大学湘雅医学院肿瘤研究所

出　　处：《中华微生物学和免疫学杂志》2002年第2期198-201,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家 973项目 (G19980 5 12 0 1);国家自然科学基金杰出青年基金 ( 395 2 5 0 2 2 );美国中华医学基金(CMB96 6 5 5 );国家自然科学基金重点项目 ( 39830 410 )

摘　　要：目的　探讨EB病毒LMP1分子致瘤机制 ,在已证实鼻咽癌细胞系中LMP1有效激活NF κB或AP 1的基础上 ,对LMP1是否通过NF κB或AP 1促进IL 8分泌进行探讨。方法　以稳定表达LMP1及其 3种突变体、空白载体的鼻咽癌细胞系 [HNE2 LMP1、HNE2 LMP1(1 185 )、HNE2 LMP1(1 2 31)、HNE2 LMP1△ 187 35 1和HNE2 pSG5 ]及antisense LMP1处理的HNE2 LMP1鼻咽癌细胞系为材料 ,将IL 8报道质粒瞬时导入这些细胞系中 ,通过测定luciferase值以反映LMP1是否促进IL 8转录 ;将mutAP 1 IL 8 luc或IκBα(S32A S36A)表达质粒导入HNE2 LMP1细胞系中 ,比较其IL 8报道活性 ,以确定LMP1是否通过AP 1或NF κB诱导IL 8转录 ;利用ELISA方法测定HNE2 LMP1、HNE2 pSG5、anti sense LMP1处理的HNE2 LMP1鼻咽癌细胞系中的IL 8浓度 ,进一步从蛋白水平上确定LMP1是否促进IL 8分泌。结果　与HNE2 pSG5相比 ,在HNE2 LMP1、HNE2 LMP1△ 187 35 1和HNE2 LMP1(1 2 31)细胞系中IL 8报道活性分别升高了原来水平的 11.5、8.6和 3.4倍 ,而HNE2 LMP1(1 185 )对IL 8报道活性不影响。在HNE2 LMP1细胞系中IL 8蛋白水平提高了 17.4倍 ,而antisense LMP1则使HNE2 LMP1细胞的IL 8报道活性及蛋白水平分别下降到原来水平的 18.3%和 9.2 % ;Objective To study the regulation of LMP1, which is a pleiotropic protein encoded by EBV, on interleukin-8 production in nasopharyngeal carcinoma cells via an NF-κB or AP-1 signaling pathway. Methods To determine whether LMP1 contributes to IL-8 production, nasopharyngeal cell lines of HNE2-LMP1, HNE2-LMP1(1-185), HNE2-LMP1(1-231), HNE2-LMP1△187-351 and HNE2-pSG5 as well as antisense-LMP1 treated HNE2-LMP1 cell line were transiently transfected with IL-8-luc plasmid, and the relative reporter activity was assessed. The supernatants of HNE2-LMP1, HNE2-pSG5 and antisense LMP1 treated HNE2-LMP1 cells were analyzed using IL-8-specific ELISA. To examine the contribution of the AP-1 site to LMP1-mediated IL-8 promoter activity, the mutated AP-1 site construct (mut AP-1/IL-8-luc) was transfected into HNE2-LMP1 cells, and the luciferase activity was examined. The role for NF-κB in IL-8 promoter regulation was demonstrated by the ability of a constitutively active IκBα mutant (IκBα S32A/S36A), which had been shown previously to interfere with LMP1-mediated NF-κB activation. Results LMP1, LMP1(1-231) or LMP1Δ187-351 mutants could induce an increase in IL-8 production compared with vector-transfected cells. However, a dramatic decrease in LMP1-induced IL-8 secretion was observed by introducing TRAF2 domain negative mutant, mut AP-1/IL-8-luc or IκBα (S32A/S36A)into HNE2-LMP1 cells. ELISA assays demonstrated that LMP1 potently enhanced IL-8 secretion, and this effect was inhibited by the introduction of antisense LMP1 into HNE2-LMP1 cells. Conclusion Epstein-Barr virus-encoded LMP1 induces IL-8 production in nasopharyngeal carcinoma cells via an NF-κB or AP-1 signaling pathway.

关 键 词：潜伏期膜蛋白1 细胞因子 信号传导 EB病毒 IL-8分泌 NF-ΚB AP-1 鼻咽癌 

分 类 号：R739.63[医药卫生—肿瘤]