Epstein‐Barr virus latent membrane protein 1 induces the matrix metalloproteinase‐1 promoter via an Ets binding site formed by a single nucleotide polymorphism: Enhanced susceptibility to nasopharyngeal carcinoma

Abstract

The Epstein-Barr Virus (EBV) latent membrane protein 1 (LMP1) has a significant role in several malignancies, including nasopharyngeal carcinoma (NPC). LMP1 is the principal oncoprotein, and we have shown that it also induces a set of factors that mediates invasion, angiogenesis and metastasis. Matrix metalloproteinase-1 (MMP1) is also involved in several malignancies. A single guanine insertion polymorphism (2G) in the MMP1 promoter creates an Ets binding site that causes high levels of transcription and correlates with risk for some malignancies. Here, we evaluate the impact of this 2G insertion type on NPC. We genotyped 44 Japanese and 39 Taiwanese NPC patients, as well as 58 Japanese and 23 Taiwanese healthy controls. The proportion of 2G homozygotes was higher in the NPC groups than in controls (Japanese: p = 0.02, odds ratio (OR) = 2.49; Taiwanese: p = 0.02, OR = 3.66). An analysis of overall survival rates in the patients with NPC, and the 1G/1G genotype disclosed a favorable prognosis (5-year survival rate = 100%, p = 0.04). Multivariate analysis showed that 1G/1G has independent prognostic significance. We also examined whether LMP1 enhances MMP1 expression in epithelial cells in culture. LMP1-transfected cells with 2G/2G genotype expressed MMP1, which was abolished by activator protein-1 (AP1) dominant-negative (DN) and Ets-DN. LMP1 also induced active MMP3, which can cleave latent MMP1, and AP1-DN and Ets-DN suppressed the MMP3 expression. These results suggest that LMP1-induced MMP1 and MMP3 are closely linked and show that LMP1 activates MMP1 via an Ets binding site formed by 2G, which is a candidate marker for both risk and prognosis of NPC. ' 2005 Wiley-Liss, Inc.Key words: SNP; MMP1; LMP1; Ets binding site; AP1EBV is a human herpes virus associated with an increasing number of malignancies, both lymphoid and epithelial in origin, including Burkitt's lymphoma and nasopharyngeal carcinoma (NPC). 1-3 Among the viral proteins expressed in the transformed cells, the nuclear antigens, Epstein-Barr nuclear antigen 2 (EBNA2), EBNA3A, EBNA3c and the integral membrane protein called latent membrane protein 1 (LMP1) are essential for growth transformation and immortalization of resting B lymphocytes. 4,5 The carboxyl-terminal portion of LMP1, which is in the cytoplasmic domain of the protein, contains 2 functional domains. The proximal domain, COOH-terminal activation region 1 (CTAR1), interacts with tumor necrosis factor receptor-associated factors and induces nuclear factor-kB (NF-kB). 6,7 The distal domain, CTAR2, is the dominant NF-kB and c-Jun N-terminal kinase (JNK) activating region of LMP1. 8,9 Previously, we showed that the CTAR1 domain also induces Ets1, a member of the Ets transcription family, and recognizes specific nucleotides sequences with a GGAA/T core sequence. We also showed that LMP1 induces c-Met, which enhances cell motility, via Ets1. [10][11][12] We have shown that LMP1 induces MMP9, a type IV collagenase that is a key enzyme in metastasis and cell invasion. 12-15 However, MMP9 is un...

Induction of Receptor for Advanced Glycation End Products by EBV Latent Membrane Protein 1 and Its Correlation with Angiogenesis and Cervical Lymph Node Metastasis in Nasopharyngeal Carcinoma

“…As previously reported, LMP1 transactivates MMP9 production through the activation of NF-nB and activator protein-1 signaling pathways (7). We have also reported that LMP1 promotes cell migration and invasive growth via Ets-1 expression in human epithelial cells (9). These results suggest that the essential binding sites for transactivation of the promoter by LMP1 are, at least in part, gene-specific in epithelial cells.…”

“…Recently, we reported that induction of Twist by LMP1 is directly associated with the metastatic nature of NPC (8). LMP1 also promotes cell migration and invasive growth via Ets-1 expression (9). In addition, LMP1 induces and causes the release of fibroblast growth factor 2 into extracellular fluid (10).…”

Genome-Wide Expression Profiling Reveals EBV-Associated Inhibition of MHC Class I Expression in Nasopharyngeal Carcinoma

“…High MMP-1 expression might contribute to the highly malignant nature of NPC. LMP1-positive tumors also overexpressed SPIB, consistent with epithelial cell studies showing that LMP1 induces MMP-1 transcription via ETS transcription factors, such as SPIB (64). In addition, another recent report described a possible link between EBV LMP2A and NPC metastasis (65).…”

Twist and Epithelial-Mesenchymal Transition Are Induced by the EBV Oncoprotein Latent Membrane Protein 1 and Are Associated with Metastatic Nasopharyngeal Carcinoma

“…NPCs with high LMP1 expression are reported to show an increased tendency toward metastasis compared with those with low LMP1 expression (12,13,(15)(16)(17)(18). Further, we have produced evidence in recent years that LMP1 can up-regulate multiple invasion, angiogenesis, and metastasis factors, and clarified the molecular mechanisms of their induction (12,13,16,17,(19)(20)(21)(22)(23)(24)(25)(26)(27).…”