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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 the effect of EBV-LMP1 on the transcriptional activation of adjacent AP 1 (5 33) and Ets (5 4 0) promoters in the promoter region of MMP 9, Regulation of MMP-9 expression in nasopharyngeal carcinoma cells through cross-talk between c-Jun and Ets1. Methods Site-directed mutagenesis was used to establish the MMP-9 promoter region of MMP-9 promoter. The binding sites of Ets (540) and AP-1 (-533) Mutants and Co-Mutants. The effects of LMP1 on the activity of these mutant reporters were compared in a tetracycline-regulated LMP1-expressing nasopharyngeal carcinoma cell line L7. The influence of c-Jun, Ets1 and their cross talk on LMP1-mediated MMP 9 expression was observed by GelatinZymography by blocking the antisense oligonucleotides against c-Jun and Ets1. Results Compared with the wild-type MMP-9-CAT plasmid, the reporter gene activity of the mutant was decreased (P <0.01) and decreased by MMP-9 -CATAP -1 (5 33) / Ets The most significant; c Jun and Ets1 antisense oligonucleotides alone or together block LMP1-mediated MMP -9 activity were reduced to co-block the role of the most obvious. Conclusion EBV-LMP1 can regulate the expression of MMP-9 in nasopharyngeal carcinoma cells through cross talk between transcription factors c-Jun and Ets1.