国家重点基础研究发展规划(973)“恶性肿瘤发生发展的基础研究”项目(G1998051201) ,国家自然科学基金(30000087和30100005)和国家自然科学基金杰出青年基金(39525022)资助项目.
This work was supported by grants from The Special Funds for Major State Basic Research of China(G1998051201), The National Natural Sciences Foundation of China(30000087, 30100005) and The National Science Fund for Distinguished Young Scholars(39525022).
探讨了EB病毒编码的潜伏膜蛋白1(LMP1)是否通过STAT3调控诱导血管内皮细胞生长因子(VEGF)的表达.利用蛋白质印迹的方法对HNE2、HNE2-LMP1以及瞬时转染STAT3显性负性突变体STAT3β的HNE2-LMP1细胞中VEGF含量进行检测,发现LMP1可以上调VEGF的表达,而STAT3β可以抑制VEGF的上调;利用LMP1可控表达细胞系tet-on-LMP1-HNE2进行LMP1时间和剂量诱导表达研究,发现VEGF可以随LMP1的动态表达而表达;将VEGF野生型报告基因和VEGF潜在的STAT3转录因子突变体报告基因与LMP1表达载体分别共转染研究发现,LMP1可以激活VEGF的转录,这种转录通过VEGF启动子区STAT3转录因子的结合位点发挥作用;电泳迁移率变动分析(EMSA)确证了STAT3的这种DNA位点的特异性活性.结果表明:EB病毒编码的LMP1 在鼻咽癌细胞中可以增加VEGF的转录和表达,并能通过VEGF启动子区STAT3转录因子结合位点发挥作用.
In order to identify whether EB virus latent membrane protein 1 (LMP1) can regulate vascular epithelial growth factor (VEGF) expression via STAT3 in nasopharyngeal carcinoma cell line, Western blotting was used to detect VEGF expression in HNE2, HNE2-LMP1 and HNE2-LMP1 transient transfected with STAT3 dominant negative plasmid STAT3β. VEGF is up-regulated by LMP1 and STAT3 dominant negative STAT3β can down-regulate VEGF expression. In tet-on-HNE2-LMP1 cell line, in which LMP1 can be tightly regulated, VEGF is expressed in dose and time dependent fashion. After cotransfected LMP1 with VEGF wild promoter luciferase reporter and 848 mutated luciferase reporter respectively, luciferase assay results indicated that LMP1 can up-regulate wild promoter luciferase activity and not that of 848 mutated luciferase reporter. Electrophoretic mobility shift assay results in HNE2 and HNE2-LMP1 cell line indicated that LMP1 enhances STAT3 binding ability in VEGF promoter. In conclusion, latent membrane protein 1 encoded by EB virus can enhance VEGF transcription and expression in nasopharyngeal carcinoma cell line via activating STAT3 binding ability in VEGF 848 promoter binding site.
谭运年,陶永光,李力力,刘素芳,唐敏,顾焕华,曹亚. EB病毒潜伏膜蛋白1在鼻咽癌细胞中通过STAT3促进VEGF表达[J].生物化学与生物物理进展,2004,31(5):427-431
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