Matrix metalloproteinases contribute to vascular remodeling by breaking down extracellular-matrix while new matrix is synthesized. Of the variety of MMPs, stromelysin-1 and gelatinase B may have key roles in coronary artery atherosclerosis. Moreover, The 5A/6A polymorphism in the promoter region of the stromelysin-1 gene may be a pathogenetic risk factor for acute myocardial infarction. Gelatinase B (92-kDa type IV collagenase and MMP-9) is one of the MMPs found to be highly expressed in the disruption-prone regions of atherosclerotic plaques. C- to T substitution at the promoter site (-1562) resulted in the higher promoter activity of the T-allelic promoter. The R279Q polymorphism in exon 6 led to the substitution of adenosine by guanine, and was a common polymorphism in the general population. We evaluated the relation between these polymorphisms and stable angina, the severity of atherosclerosis in coronary artery disease, and instent restenosis after percutaneous coronary angioplasty. The study population was composed of 131 patients with stable angina (mean age 61.3 years, 89 males) and 117 control subjects (mean age 59.3 years, 59 males). Coronary angiographies were performed in all cases at Yonsei University Cardiovascular Hospital from February 1998 to June 2000. The genotype for each polymorphism was determined using a SNaPshotTM kit and by restriction fragment length polymorphism (RFLP). The prevalence of 5A containing a polymorphism of the stromelysin-1 gene was higher in the stable angina group than in control patients, but no difference in the two polymorphisms of the gelatinase B gene was found between the two groups. By multiple logistic analysis, the 5A-allele of the stromelysin-1 gene was found to be an independent risk factor of stable angina with an odds ratio of 2.29 (95% CI; 1.19-4.38). However, the severity of atherosclerosis in coronary artery or in stent restenosis was not related to any polymorphism of stromelysin-1 or gelatinase B. Our results show that functional genetic variation of stromelysin-1 could be a significant risk factor for stable angina, and might play an important role in coronary atherosclerosis involving vascular remodeling.
Aged ; Angina Pectoris/*etiology/genetics ; Coronary Restenosis/etiology/genetics ; Female ; Gelatinase B/*genetics ; Genotype ; Human ; Male ; Middle Age ; *Polymorphism (Genetics) ; Promoter Regions (Genetics) ; Stromelysin 1/*genetics

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Animal ; Blotting, Northern/methods ; Brain/pathology ; Brain Neoplasms/pathology ; Brain Neoplasms/enzymology* ; Enzyme Activation ; Gelatinase A/metabolism ; Gelatinase A/genetics* ; Gelatinase B/metabolism ; Gene Expression Regulation, Enzymologic* ; Glioma/pathology ; Glioma/enzymology* ; Human ; Metalloendopeptidases/genetics* ; Papio ; Tissue Inhibitor-of Metalloproteinase-2/genetics ; Tissue-Inhibitor of Metalloproteinase-1/genetics ; Tumor Cells, Cultured

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Animal ; Blotting, Northern/methods ; Brain/pathology ; Brain Neoplasms/pathology ; Brain Neoplasms/enzymology* ; Enzyme Activation ; Gelatinase A/metabolism ; Gelatinase A/genetics* ; Gelatinase B/metabolism ; Gene Expression Regulation, Enzymologic* ; Glioma/pathology ; Glioma/enzymology* ; Human ; Metalloendopeptidases/genetics* ; Papio ; Tissue Inhibitor-of Metalloproteinase-2/genetics ; Tissue-Inhibitor of Metalloproteinase-1/genetics ; Tumor Cells, Cultured

Expression of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9), which correlates with tumor invasion and metastasis, has been known to be regulated by several intracellular signaling pathways. Since the CD9 membrane protein has been implicated in signal transduction and malignant progression of cancer cells, we examined the functional involvement of CD9 in the regulation of MMP-2 and MMP-9 expression by using stable CD9 transfectant clones of MelJuso human melanoma cells. The CD9 cDNA-transfected cells with elevated CD9 expression displayed increased MMP-2 and decreased MMP-9 expression when compared with the mock transfectant cells. Among several signal pathway inhibitors tested, SB203580 and SP600125, which inhibit p38 MAPK and JNK respectively, completely blocked the CD9-stimulated MMP-2 expression. Phosphorylation levels of p38 MAPK and c-Jun in MelJuso cells were also significantly increased by CD9 transfection. In addition, the down-regulation of p38 MAPK and JNK by siRNA transfection resulted in a decrease in MMP-2 expression by MelJuso cells. Promoter analysis and gel shift assay showed that the CD9-induced MMP-2 expression is mediated by a functional AP-1 site through interactions with AP-1 transcription factors including c-Jun. These results suggest that CD9 induces MMP-2 expression by activating c- Jun through p38 MAPK and JNK signaling pathways in human melanoma cells.
Antigens, CD/*metabolism ; Electrophoretic Mobility Shift Assay ; Enzyme Activation ; Gelatinase A/genetics/*metabolism ; Gelatinase B/metabolism ; Humans ; JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors/genetics/*metabolism ; Melanoma/*metabolism/pathology ; Membrane Glycoproteins/*metabolism ; Promoter Regions (Genetics) ; Proto-Oncogene Proteins c-jun/*metabolism ; RNA, Small Interfering/pharmacology ; Research Support, Non-U.S. Gov't ; *Signal Transduction ; Skin Neoplasms/metabolism/pathology ; Transcription Factor AP-1/metabolism ; Transfection ; p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors/genetics/*metabolism

Kawasaki disease (KD) is a major cause of acquired coronary artery diseases in childhood. The serum levels of matrix metalloproteinase (MMP)-3 and MMP-9 in KD have been reported to be significantly higher than other diseases. Several studies have demonstrated that MMP-3 5A/6A polymorphism and MMP-9 C-1562T polymorphism modify each transcriptional activity in allele specific manner. We hypothesized that these polymorphisms may play a role as a risk factor for development of coronary artery lesions (CAL) in KD. Eighty-three patients, diagnosed with KD in Cheju National University Hospital from January 2000 to February 2004, were divided into two groups according to the presence of CAL. Genotyping of MMP-3 and MMP-9 gene polymorphisms were determined by restriction fragment length polymorphism. With regard to MMP-3 gene polymorphism, the KD with CAL group had a higher frequency of 6A/6A genotype than control group (p=0.0127) and the KD without CAL group (p=0.0036). However, no significant differences in the allele and genotype distributions of the MMP-9 polymorphism were observed. These findings suggest that MMP-3 6A/6A genotype may be an independent risk factor for CAL formation in KD.
Adolescent ; Adult ; Aged ; Alleles ; Child ; Child, Preschool ; Coronary Arteriosclerosis/enzymology/etiology/*genetics ; Female ; Gelatinase B/genetics ; Gene Frequency ; Genotype ; Humans ; Infant ; Male ; Middle Aged ; Mucocutaneous Lymph Node Syndrome/*complications ; *Polymorphism, Genetic ; Promoter Regions (Genetics)/*genetics ; Research Support, Non-U.S. Gov't ; Risk Factors ; Stromelysin 1/*genetics

Increased production of matrix metalloproteinases (MMPs) has been associated with increases in invasive and metastatic potential in many types of human carcinoma. Tissue inhibitors of metalloproteinase (TIMP)-1 inhibits most interstitial collagenases and MMP-9. TIMP-2 binds specifically and noncovalently to the pro-form of MMP-2 and inhibits its enzyme activity. In this study, we examined TIMP-1 and TIMP-2 expressions in relation to clinicopathological variables in colorectal carcinoma with in situ hybridization and immunohistochemistry. TIMP-1 and TIMP-2 expressions were localized overwhelmingly to pericancer stromal cells, while malignant and normal mucosal cells were weak or negative. Strong stromal TIMP-1 immunoreactivity correlated with Dukes' stage (p=0.022), status of lymph node metastasis (p=0.044) and poor survival (p= 0.005). The degree of immunohistochemical staining of TIMP-2 did not correlate with all clinicopathological variables. The correlation between enhanced TIMP-1 expression and advanced stage and poor survival suggest a growth promoting activity of TIMP-1 in colorectal carcinoma.
Adenocarcinoma/pathology ; Adenocarcinoma/mortality ; Adenocarcinoma/enzymology* ; Adult ; Aged ; Aged, 80 and over ; Antibodies ; Collagenases/immunology ; Collagenases/genetics* ; Collagenases/analysis ; Colorectal Neoplasms/pathology ; Colorectal Neoplasms/mortality ; Colorectal Neoplasms/enzymology* ; DNA Probes ; Female ; Gelatinase A ; Gelatinase B ; Gelatinases/immunology ; Gelatinases/genetics* ; Gelatinases/analysis ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Human ; In Situ Hybridization ; Male ; Metalloendopeptidases/immunology ; Metalloendopeptidases/genetics* ; Metalloendopeptidases/analysis ; Middle Age ; Predictive Value of Tests ; RNA, Messenger/analysis ; Stromal Cells/pathology ; Stromal Cells/enzymology ; Survival Analysis ; Tissue Inhibitor-of Metalloproteinase-2/immunology ; Tissue Inhibitor-of Metalloproteinase-2/genetics* ; Tissue Inhibitor-of Metalloproteinase-2/analysis ; Tissue-Inhibitor of Metalloproteinase-1/immunology ; Tissue-Inhibitor of Metalloproteinase-1/genetics* ; Tissue-Inhibitor of Metalloproteinase-1/analysis

During chronic inflammatory response, mono- cytes/macrophages produce 92-kDa matrix metalloproteinase-9 (MMP-9), which may contribute to their extravasation, migration and tissue remodeling. Activation of peroxisome proliferator- activated factor receptor-gamma (PPAR-gamma) has been shown to inhibit MMP-9 activity. To evaluate whether ox-LDL, a PPAR-gamma activator, inhibits PMA-induced MMP-9 expression and activity, and if so, whether CD36 and PPAR-gamma are involved in this process, we investigated the effect of ox-LDL on MMP-9 expression and activity in PMA-activated human monocytic cell line U937. PMA-induced MMP-9 expression and activity were suppressed by the treatment with ox-LDL (50 micrigram/ml) or PPAR-gamma activators such as troglitazone (5 micrometer), ciglitazone (5 micrometer), and 15d- PGJ2 (1 micrometer) for 24 h. This ox-LDL or PPAR-gamma activator-mediated inhibition of micrometer P-9 activity was diminished by the pre-treatment of cells with a blocking antibody to CD36, or PGF2a (0.3 micrometer), which is a PPAR-gamma inhibitor, as well as overexpression of a dominant-negative form of CD36. Taken together, these results suggest that ox-LDL suppresses PMA-induced MMP-9 expression and activity through CD36-mediated activation of PPAR-gamma.
Antibodies, Blocking/pharmacology ; Antigens, CD36/immunology/*physiology ; Cells, Cultured ; Chromans/pharmacology ; Gelatinase B/antagonists & inhibitors/genetics/*metabolism ; Humans ; Lipoproteins, LDL/pharmacology/*physiology ; Monocytes/drug effects/*enzymology/metabolism ; NF-kappa B/antagonists & inhibitors ; PPAR gamma/*metabolism ; Prostaglandin D2/*analogs & derivatives/pharmacology ; RNA, Messenger/analysis/metabolism ; Research Support, Non-U.S. Gov't ; Tetradecanoylphorbol Acetate/antagonists & inhibitors/pharmacology ; Thiazolidinediones/pharmacology ; Transcription, Genetic/drug effects