The expression of matrix metalloproteinase 9 is enhanced by Epstein–Barr virus latent membrane protein 1

Matrix metalloproteinases (MMPs) are frequently expressed in malignant tumor cells and are thought to play crucial roles in tumor invasion and metastasis. Here we report that expression of MMP9 is increased in Epstein-Barr virus (EBV)-infected type III latency lymphoma cell lines, but not in type I lines where latent viral gene expression is highly restricted. Type III cell lines express abundant EBV latent membrane protein 1 (LMP1), the principal EBV oncoprotein, as well as the other latency proteins including the transcriptional factor, EBV nuclear antigen 2, which is also required for cell immortalization. Transfection of an LMP1 expression plasmid in the C33A cell line increased MMP9 expression, whereas overexpression of EBV nuclear antigen 2 did not. Three motifs, homologous to the binding sites of NF-kappaB, SP-1, and AP-1 proteins, contribute to induction of the MMP9 promoter by 12-O-tetradecanoyl-phorbol-13-acetate and tumor necrosis factor alpha. Here we report that binding sites for NF-kappaB, SP-1, and AP-1 also contribute to induction of the MMP9 promoter by the viral protein, LMP1, mainly through the NF-kappaB and, to a lesser extent, the SP-1 and AP-1 sites. Moreover the AP-1 binding site is essential in that mutation of it abolished reporter gene induction by LMP1. The enhancement of MMP9 expression was blocked by cotransfection of an IkappaB expression plasmid. Thus in addition to its transforming properties, the oncoprotein LMP1 may contribute to invasiveness and metastasis of EBV-associated tumors such as nasopharyngeal carcinoma.