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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 % ;导入m
Objective To investigate the mechanism of tumorigenesis of Epstein-Barr virus (LMP1), and to investigate whether LMP1 promotes IL-8 secretion through NF-κB or AP-1 on the basis that LMP1 effectively activates NF-κB or AP-1 in nasopharyngeal carcinoma cell lines. Methods HNE2 LMP1, HNE2 LMP1 (1 185), HNE2 LMP1 (1 2 31), HNE2 LMP1 △ 187 35 1 and HNE2 pSG5 were stably expressed in LMP1 and its 3 mutants. antisense LMPl -treated HNE2 LMPl nasopharyngeal carcinoma cell line was used as material, IL8 reporter plasmids were transiently introduced into these cell lines by measuring the luciferase value to reflect whether LMPl promoted IL8 transcription; mutAP1 IL8 luc or IκBα (S32A S36A ) Expression plasmid was introduced into HNE2 LMP1 cell line to compare its IL 8 reporter activity to determine whether LMP1 induces IL 8 transcription via AP 1 or NF κB; HNE2 LMP1, HNE2 pSG5, anti sense LMP1-treated HNE2 LMP1 nasal The IL8 concentration in pharyngeal cancer cell lines further determines from the protein level whether LMP1 promotes IL8 secretion. Results Compared with HNE2 pSG5, the reporter activity of IL 8 in the HNE2 LMP1, HNE2 LMP1 △ 187 35 1 and HNE2 LMP1 (1231) cell lines increased by 11.5, 8.6 and 3.4 times of the original level respectively, while the HNE2 LMP1 ( 1 185) No effect on IL 8 reporter activity. In the HNE2 LMP1 cell line, the level of IL8 protein was increased by 17.4 times, while antisense LMP1 reduced the IL8 reporter activity and protein level of HNE2 LMP1 cells to 18.3% and 9.2%