High levels of MMP‐1 expression in the absence of the 2G single nucleotide polymorphism is mediated by p38 and ERK1/2 mitogen‐activated protein kinases in VMM5 melanoma cells

Abstract

Matrix metalloproteinase-1 (MMP-1) is one of only a few enzymes with the ability to degrade the stromal collagens (types I and III) at neutral pH, and high expression of MMP-1 has been associated with aggressive and invasive cancers. We recently reported a single nucleotide insertion/deletion polymorphism (SNP) in the collagenase-1 (MMP-1) promoter (Rutter et al. [1998] Can. Res. 58:5321-5325), where the insertion of an extra guanine (G) at -1607 bp creates the sequence, 5'-GGAA-3 (2G allele), compared to the sequence 5'-GAA-3' (1G allele). The presence of 2G constitutes a binding site for the ETS family of transcription factors, and increases MMP-1 transcription in fibroblasts and A2058 melanoma cells cultured in vitro. In addition, the presence of the 2G allele has been linked to several aggressive malignancies as well as to enhanced expression of MMP-1. In this study, we describe a melanoma cell line, VMM5, that is 1G homozygous, but that is invasive and expresses high levels of MMP-1 constitutively. The high level of MMP-1 expression in VMM5 cells is due to the utilization of both the p38 and ERK1/2 transduction pathways. In contrast, in the A2058 cell line, which also expresses MMP-1 constitutively and which is 2G homozygous, only the ERK pathway is activated. Thus, our data suggest that in the absence of 2G allele and in the presence of the appropriate transcription factors, tumor cells may use alternative signal/transduction pathways and cis-acting sequences to achieve high levels of MMP-1 expression, which contribute to the ability of tumor cells to invade, regardless of their genotype.

Absence of a Correlation between the Presence of a Single Nucleotide Polymorphism in the Matrix Metalloproteinase 1 Promoter and Outcome in Patients of Chondrosarcoma

“…This suggests that Ets family proteins and partner proteins may differ in various cell types (32). Alternatively, other pathways regulating MMP-1 expression may act independently of the SNP at Ϫ1607 bp (33). Although various human tumor tissues have demonstrated coexpression of Ets factor and MMPs, there was no correlation between Ets expression and metastasis in pancreatic and thyroid carcinoma (34,35).…”

Topical Treatment with Inhibitors of the Phosphatidylinositol 3′-Kinase/Akt and Raf/Mitogen-Activated Protein Kinase Kinase/Extracellular Signal-Regulated Kinase Pathways Reduces Melanoma Development in Severe Combined Immunodeficient Mice

“…Because BRaf mutation activates the mitogen-activated protein kinase cascade, and loss of PTEN activates PI3K, these observations suggest that these two pathways are important mediators of melanoma downstream of Ras. The PI3K/Akt and Raf/MEK/Erk pathways promote tumorigenesis essentially by positively regulating cell survival, cell cycle progression (27)(28)(29), tumor angiogenesis (30 -32), and tumor invasion (33)(34)(35).…”

Overexpression of Collagenase 1 (MMP-1) Is Mediated by the ERK Pathway in Invasive Melanoma Cells

“…Extracts were heated for 10 min at 100°C and resolved by SDS-PAGE and immunoblotted as described previously (16,18). Antibodies to p38, pp38, MEK1/2, ppMEK1/2, pp42/44, and p42/44 were purchased from Cell Signaling.…”

“…RT and real-time PCR were performed using protocols and reagents from Applied Biosystems Taqman RT reagent kit and Sybr Green PCR master mix as described (16). Briefly, 2 g of DNase-treated RNA was reverse-transcribed in a 20-l reaction containing 5.5 mM MgCl 2 , 500 M each dNTP, 2.5 M oligonucleotide d(T) 16 , 0.4 unit/l RNase inhibitor, and 1.25 unit/l Multiscribe reverse transcriptase. The reactions were incubated at 25°C for 5 min, 48°C for 30 min, and then 95°C for 5 min.…”