EB病毒潜伏膜蛋白1介导鼻咽癌细胞中转录因子Ets-1表达的实验研究  

EB virus encoded latent membrane protein 1 mediated expression of transcription factor Ets-1 in nasopharyngeal carcinoma cells

作　　者：曾亮[1] 刘轶平[1] 王海[1] 陶永光[1] 赵晓荣[1] 李嵬[1] 曹亚[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所

出　　处：《中华肿瘤杂志》2004年第8期454-457,共4页Chinese Journal of Oncology

基　　金：国家重点基础发展规划 ( 973 )资助项目 ( 2 0 0 2CB5 13 10 0 );国家自然科学基金资助项目 ( 3 0 10 0 0 0 5 )

摘　　要：目的　探讨鼻咽癌 (NPC)细胞中EB病毒潜伏膜蛋白 1(LMP1)是否可介导转录因子Ets 1的表达。方法　采用受四环素 (Dox)调控的、LMP1表达的NPC细胞系 (pTet on LMP1HNE2 ) ,用Dox诱导LMP1表达 ,并检测Ets 1的表达。采用逆转录PCR(RT PCR)方法检测Ets 1RNA水平的变化 ;应用Westernblot方法检测Ets 1蛋白质表达情况 ;应用免疫共沉淀和Westernblot方法检测Ets 1磷酸化水平 ;应用EMSA方法检测Ets 1的DNA结合活性。结果　在不同诱导时间、不同浓度Dox诱导的pTet on LMP1HNE2细胞中 ,LMP1表达与Ets 1RNA表达、蛋白质表达、苏氨酸磷酸化水平及DNA结合活性在一定范围内呈正相关。结论　LMP1可介导NPC细胞中Ets 1的转录活化和表达 。Objective To elucidate the expression of tanscription factor Ets-1 mediated by EB virus encoded latent membrane protein 1 (LMP1) in nasopharyngeal carcinoma (NPC) cells. Methods LMP1-expressing NPC HNE2 cells regulated by Tet-on system (pTet-on-LMP1 HNE2) were used. Expression of LMP1 and Ets-1 was observed after induction with Doxycycline (Dox). Expression of Ets-1 mRNA and protein was detected by RT-PCR and Western blot, respectively. The phosphorylation level of Ets-1 protein was examined by co-immunoprecipitation. The DNA binding activity of Ets-1 was detected by electrophoretic-mobility shift assay (EMSA). Results After induction with Dox in pTet-on-LMP1 HNE2 cells, to some extent, the expression of Ets-1 mRNA and protein , its phosphorylation level and DNA binding activity were increased with enhancement of LMP1 expression. Conclusion LMP1 induces transcriptional activation and expression of Ets-1 which may contribute to the development of NPC.

关 键 词：EB病毒 膜蛋白1 鼻咽癌 癌细胞 NPC 转录因子Ets-1表达 爱泼斯坦-巴尔病毒 肿瘤 

分 类 号：R739.63[医药卫生—肿瘤]