【摘要】 【目的】检测EB病毒潜伏膜蛋白1(LMP1)远端启动子L1-TR在鼻咽癌(NPC)组织中的突变情况并探讨突变对启动子活性的影响。【方法】用聚合酶链反应(PCR)方法分别扩增出B95.8细胞和新鲜鼻咽癌组织细胞中EB病毒(EBV)的L1-TR片段,并测序比较其核苷酸序列的差异。构建含原型和突变型L1-TR启动子的荧光素酶报告质粒并分别转染HaCat细胞、B95.8细胞和C666-1细胞,比较两种启动子的活性。【结果】和B95.8原型比较,鼻咽癌组织中的EB病毒L1-TR启动子区域有多处缺失、插入和点突变,且启动子活性明显降低(P<0.05)。【结论】鼻咽癌组织中LMP1启动子L1-TR的活性与B95.8原型相比明显降低,这在EB病毒的免疫逃避机制中可能有一定的意义。更多还原

【Abstract】 [Objective] To investigate the mutation of latent membrane protein 1(LMP1) distal promoter L1-TR in nasopharyngeal carcinoma(NPC) tissue and the influence on promoter activity.[Methods] The L1-TR fragments of Epstein-Barr virus(EBV) genome in B95.8 cells and fresh NPC tissue cells were amplified by using polymerase chain reaction(PCR) and the difference between their nucleotides sequence were compared by direct sequencing.The luciferase reporter containing mutant or prototype L1-TR promoter was constructed and transfected into the HaCat,B95.8,and C666-1 cells respectively to compare the transcriptional activity.[Results] The nucleotides insertion,deletion and point mutations were found in L1-TR promoter derived from NPC tissue compared with B95.8 prototype,and activity of the mutant L1-TR promoter was reduced markedly(P < 0.05).[Conclusions] The activity of the LMP1 promoter in NPC tissue is significantly lower than that of B95.8 prototype L1-TR,which might contribute to the immune evasion mechanism of EBV.更多还原