EB病毒编码潜伏膜蛋白1通过转录因子c-Jun和Ets1间信息交流调控鼻咽癌中基质金属蛋白酶9的表达  

Cross-talk between c-Jun/Ets1 involved in EB virus-encoded latent membrane protein 1 regulates expression of matrix metalloproteinase-9 in nasopharyngeal carcinoma

作　　者：曾亮[1] 刘轶平[1] 陶永光[1] 艾米丹[1] 赵晓荣[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》2005年第4期204-208,共5页Chinese Journal of Oncology

基　　金：国家重点基础发展规划资助项目 (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　探讨在EBV -LMP1作用下,基质金属蛋白酶9(MMP 9)启动子区相邻的AP 1( 5 33)和Ets( 5 4 0 )结合位点对其转录活化的影响,并确定LMP1可通过c -Jun和Ets1间信息交流(cross -talk)调控鼻咽癌细胞中MMP 9的表达。方法　应用定点突变技术,在野生型MMP- 9 -CAT质粒的基础上,建立MMP- 9启动子区相邻Ets( 5 4 0 )结合位点和AP- 1 (- 5 33)结合位点单独突变体和共同突变体;在四环素调控表达LMP1的鼻咽癌细胞系L7中,比较LMP1对这些突变体报道基因活性的影响。通过针对c- Jun、Ets1的硫代反义寡核苷酸进行阻断,采用GelatinZymography观察c -Jun、Ets1及其cross- talk对LMP1介导的MMP 9表达的影响。结果　与野生型MMP -9 -CAT质粒比较,突变体的报道基因活性均降低(P <0 .0 1 ) ,以MMP- 9 -CATAP -1 ( 5 33) /Ets( 5 4 0 )mt降低最为显著;在c Jun和Ets1反义寡核苷酸单独或共同阻断后,LMP1介导的MMP -9活性均降低,以共同阻断的作用最为明显。结论　EBV -LMP1可以通过转录因子c -Jun和Ets1间cross talk调控鼻咽癌细胞中MMP 9的表达。Objective To investigate effect of AP-1 and Ets binding site adjacent to matrix metalloproteinase-9(MMP-9) promoter on activation of MMP-9 transcription of nasopharyngeal carcinoma cells transfected with EBV-encoded latent membrane protein1(LMP1), and to ascertain if cross-talk between c-Jun and Ets1 is involved in LMP1-regulating expression of MMP-9. Methods Site-directed mutagenesis technique was used to establish a series of mutants, including MMP-9-CAT-Ets(-540)mt, MMP-9-CAT-AP-1(-533)mt and MMP-9-CAT-AP-1(-533)/Ets(-540)mt. After the mutants were transfected into LMP1-expressing NPC HNE2 cells regulated by Tet-on system (pTet-on-LMP1 HNE2), CAT activity of these mutants were assayed with induction of LMP1. With blockade of c-Jun or Ets1 antisense oligonucleotides, the activity of MMP-9 induced by LMP1 was assayed with gelatin zymography. Results The CAT activity of MMP-9-Ets(-540)mt-CAT, MMP-9-AP-1(-533)mt-CAT, MMP-9-AP-1(-533)/Ets(-540) mt-CAT decreased significantly compared to MMP-9-CAT wt. After blockade with c-Jun or Ets1 antisense oligonucleotides, activity of MMP-9 induced by LMP1 decreased significantly, especially with combined blockade of c-Jun and Ets1. Conclusion The results suggest that transcription factor AP-1 and Ets play an crucial role in activation of MMP-9 transcription induced by LMP1, and cross-talk between c-Jun/Ets1 is involved in expression of MMP-9 mediated by LMP1.

关 键 词：转录因子C-JUN 基质金属蛋白酶9 信息交流 潜伏膜蛋白1 EB病毒编码 Ets EBV-LMP1 转录因子C-JUN 硫代反义寡核苷酸 l反义寡核苷酸 MMP-9活性 结合位点 定点突变技术 鼻咽癌细胞系 启动子区 基因活性 突变体 方法应用 调控表达 

分 类 号：R739.63[医药卫生—肿瘤]