Promoter Region Of MMP-9 Gene Research Articles

Matrix metalloproteinase-9-1562 C/T promoter gene polymorphism in chronic periodontitis: A cross-sectional observational study.

Background:Matrix metalloproteinases (MMPs) are a major group of enzymes, released in inflamed periodontal tissues in large quantities, resulting in connective tissue matrix breakdown. One of the most predominant MMPs is MMP-9. Association between chronic periodontitis (CP) and MMP-9 gene polymorphism (GP) in some ethnic populations has been already established. The aim of the current study was to assess the association of single-nucleotide polymorphism in the promoter region of MMP-9 gene with CP in Kolkata population, if any.Materials and Methods:Forty eligible individuals were recruited for the study and grouped as A and B: Group A (twenty periodontally healthy individuals) and Group B (twenty CP patients). Venous blood samples collected from all the forty individuals were used to isolate the genomic DNA. GP for MMP-9 gene was detected using polymerase chain reaction and sequencing method. The data obtained were then analyzed statistically.Results:A significantly different genotypic and allelic variation was observed in the control and test groups, with C/T genotype and T allele present in only CP group.Conclusion:Within the limitations of this study, we can conclude that there was no significant association between patients with periodontitis and MMP-9-1562 C/T promoter GP.

Matrix Metalloproteinase-9 Gene Polymorphisms in South-West Iranian Multiple Sclerosis (MS) Patients

Background: Matrix metalloproteinases (MMPs) play important role in Multiple Sclerosis (MS), an autoimmune and neuro-inflammatory disease. It has been demonstrated that two important polymorphisms, the –1562 C/T and microsatellite (CA)n near the –90 position in the promoter region of MMP-9 gene are associated with increased MMP-9 expression, through influencing transcriptional process in MS, also some autoimmune diseases as well as cancers. In this study, the possible association of these gene polymorphisms with MS susceptibility, severity as well as age of onset in samples from south-west of Iran were examined. Methods: Polymorphisms for two regions, –1562 C/T (rs3918242) and –90 (CA)n repeat was analyzed in 170 south-west Iranian patients with MS and 200 healthy controls. Results: Results suggested that a genetic polymorphism of the MMP-9 promoter region may influence the susceptibility to MS. Significant increase frequencies of the –1562 T and long (CA)n alleles were found in patients with MS compared to controls. Carriers of two polymorphisms were younger age at disease onset. The disability status of patients, moreover, has been consistently increased by the association of T variants. Conclusion: This study suggests that genotypes and alleles of MMP-9 –1562 C/T gene as well as (CA)n polymorphisms are associated with higher MS susceptibility due to increased MMP-9 protein levels. This finding, therefore, supports the hypothesis of a significant role of this gene in the disease.

Impact of Matrix Metalloproteinase 9 on COPD Development in Polish Patients: Genetic Polymorphism, Protein Level, and Their Relationship with Lung Function.

Chronic obstructive pulmonary disease (COPD) is characterized by a decline of lung function and symptoms such as chronic bronchitis and emphysema leading from lung tissue destruction. Increased activity of matrix metalloproteinases (MMPs) and an imbalance between MMPs and their tissue inhibitors (TIMPs) are considered as factors influencing the pathogenesis of COPD. We investigated the role of genetic polymorphism and expression level of MMP-9 and concentration of its complexes with TIMPs in the development of COPD among Polish patients. We analyzed SNP in the promoter region of MMP-9 gene (rs3918242) using PCR-RFLP method among 335 COPD patients and 309 healthy individuals. Additionally, 60 COPD patients and 61 controls were tested for copy number variants (CNV) of MMP-9 (by quantitative real-time PCR) and serum levels of MMP-9 and its complexes with TIMP1 and TIMP2 (using ELISA). All subjects were analyzed for lung function using spirometry (FEV1% and FEV1/FVC parameters). We observed that allele and genotype frequencies of the SNP rs3918242, as well as the number of gene copies, were similar in COPD patient and controls groups. Serum levels of MMP-9 and MMP-9/TIMP1 complex were significantly higher in COPD patients in comparison to controls groups, although independently of analyzed gene polymorphisms. Additionally, the significant inverse relationships between parameters of lung function (FEV1% and FEV1/FVC) and proteins level were found in ridge regression models, especially we found that FEV1% decreased when MMP-9 level increased in controls and patients with COPD group. In conclusion, we found that COPD patients were predisposed to produce more MMP-9 and MMP-9/TIMP1 complex than healthy individuals. This phenomenon is probably associated with the disease-related lung environment but not with genetic features of the MMP-9.

Association of MMP-9 C-1562T polymorphism with risk of Henoch-Schönlein purpura nephritis in children of southeast China

Henoch-Schonlein purpura nephritis (HSPN) is the most prevalent vasculitis in childhood worldwide. Although the etiology of HSPN is not yet fully elucidated, it is believed to be closely related with infections and genetic predispositions. In this study, a cohort of children from southeast China, including 108 healthy controls and 184 patients were involved in. We scanned the promoter region and the whole gene for genetic variations. A C/T transition located in the promoter region of MMP-9 gene, C-1562T was identified. Subjects were genotyped by using SNaPshot sequencing and the frequencies of CC, CT and TT genotypes were 43, 33 and 24% in patients and 53, 37 and 10% in healthy controls. Allele distribution was found to be in Hardy–Weinberg equilibrium (P = 0.17). Association analysis revealed that the polymorphism was significantly related with HSPN risk (P < 0.05) and T allele was shown to be a risk factor (P = 0.003, odds ratio (OR) 1.63, 95% confidence interval (CI) 1.13–2.34). Additionally, we measured the serum MMP-9 concentration and quantified the mRNA expression. Results showed that an elevated protein level was observed in patients compared with controls in both ELISA (P = 0.0029, 136.5 μg/L vs. 103.9 μg/L) and Western blot assays. Furthermore, we also detected 1.93-fold increase in the mRNA transcription in patients carrying homozygous TT genotype, indicating enhanced promoter activity caused by the polymorphism. Based on the incidence risk, a cut-off value of serum MMP-9 served as HSPN diagnosis was suggested at 155 μg/L. Conclusively, our findings suggested that MMP-9 polymorphism was significantly associated with HSPN susceptibility, and C-1562T T allele contributed to HSPN development.

Differential Expression of Matrix Metalloproteinase-9 Gene in Wounds of Type 2 Diabetes Mellitus Cases With Susceptible -1562C>T Genotypes and Wound Severity.

Coordinated extracellular matrix deposition is a prerequisite for proper wound healing which is mainly orchestrated by matrix metalloproteinases (MMPs). Diabetic wounds generally show compromised wound healing cascade and abnormal MMP9 concentration is one of the cause. Our group have recently shown that the polymorphism -1562 C>T in the promoter region of MMP9 gene is associated with pathogenesis of wound healing impairment in T2DM patients. In present study we have done expression profiling of MMP9 gene in the wound biopsy of DFU cases. Expression level of MMP9 mRNA was then compared with susceptible -1562 C>T genotypes (TT and CT) as well as with different grades of wounds. We also screened the promoter region of MMP9 gene to see the methylation state of CpGs present there. Our study suggests that levels of MMP9 mRNA increase significantly with the wound grades. Moreover, the MMP9 levels in diabetic wounds were also dependent on -1562 C>T polymorphism in the promoter region of MMP9. Diabetic wounds also showed a significant unmethylated status of MMP9 promoter compared to control wounds. In conclusion, The risk genotypes of -1562 C>T polymorphism along with lack of methylation of CpG sites in MMP9 gene promoter may result in altered expression of MMP9 in wounds of T2DM cases resulting into nonhealing chronic ulcers in them.

A Functional Single Nucleotide Polymorphism -1562C&gt;T in the Matrix Metalloproteinase-9 Promoter Is Associated With Type 2 Diabetes and Diabetic Foot Ulcers

Impaired neovascularization is the hallmark of type 2 diabetes, which results in various macro- and microvascular complications and the development of foot ulcerations later in life. Matrix metalloproteinases (MMPs) are the key enzymes which influence matrix remodeling. Here, we aim to investigate that whether single nucleotide polymorphism (SNP -1562C>T) (rs3918242) in the promoter region of MMP-9 gene, which alters the transcriptional activity of MMP-9 is associated with type 2 diabetes and diabetic foot ulcers (DFUs). This case-control study was composed of 730 individuals, out of which 463 patients were with type 2 diabetes mellitus (T2DM) and 267 were nondiabetic healthy controls (non-DM controls). T2DM patients were subclassified as 149 cases without any secondary complications (T2DMNSC), 110 with DFUs, 204 T2DM patients having one or the other secondary complications. Genotyping for -1562C>T SNP in MMP-9 gene was done by polymerase chain reaction-restriction fragment length polymorphism method and sequencing. SNP -1562C>T of MMP-9 gene showed a significant association with T2DM and DFU. The allele distribution differed significantly between patients and normal control group (odds ratio = 1.82, P = .00005, 95% confidence interval = 1.36-2.42 for T2DM vs control and odds ratio = 2.112, P = .00048, 95% confidence interval = 1.38-3.126 for DFU vs control) indicating strong association of SNP -1562C>T of MMP-9 gene with T2DM and DFU in an Indian population. SNP -1562C>T in the promoter of the MMP-9 gene results in increased expression at the level of the transcription. To the best of our knowledge, this is the first report that suggests that SNP -1562C>T in the promoter of the MMP-9 gene is associated with T2DM and DFU. An increased MMP-9 production from high expressing T allele may promote matrix degradation.

Matrix metalloproteinase (MMP)-9 genotypes and haplotypes in preeclampsia and gestational hypertension

BackgroundAbnormal production of matrix metalloproteinases (MMPs), especially MMP-9, may play a role in hypertensive disorders of pregnancy. These alterations may result from functional genetic polymorphisms in the promoter region of MMP-9 gene, which are known to change MMP-9 expression. We examined whether 2 MMP-9 polymorphisms (C−1562T and (CA)n) and haplotypes are associated with preeclampsia and/or gestational hypertension. MethodsWe studied 476 pregnant women: 176 healthy pregnant (HP), 146 pregnant with gestational hypertension (GH), and 154 pregnant with preeclampsia (PE). Genomic DNA was extracted from whole blood and genotypes for C−1562T and (CA)n polymorphisms were determined by PCR-RFLP. Haplotype frequencies were inferred using the PHASE ver. 2.1 program. ResultsFor the g.−90(CA)13–25 polymorphism, no significant differences were found in genotype and allele distributions when PE or GH groups were compared with HP group. However, the CT genotype and T allele for g.−1562C>T polymorphism were more commonly found in GH subjects compared with the HP group (both P<0.05). Conversely, we found no differences in genotypes or allele distributions for the g.−1562C>T polymorphism when the PE and the HP groups were compared. No significant differences were found in overall distributions of haplotype frequencies when the GH or the PE group was compared with the HP group. ConclusionsThe C−1562T polymorphism in MMP-9 gene is associated with gestational hypertension, but not with preeclampsia. These findings may help to explain the higher plasma MMP-9 levels previously reported in GH compared with HP.

Association of Matrix Metalloproteinase‐9 Promoter Gene Polymorphism With Chronic Periodontitis

Matrix metalloproteinases (MMPs) are related to tissue destruction and remodeling events in periodontal diseases. A single nucleotide polymorphism in the promoter region of human MMP-9 gene is associated with the risk of some inflammatory diseases. Therefore, the aim of this study was to investigate the association between MMP-9 promoter polymorphism and severe generalized chronic periodontitis in a Turkish population. Samples of venous blood and DNA were obtained from 70 severe generalized chronic periodontitis patients and 70 healthy subjects. The alleles of the C/T polymorphism at position -1562 in the promoter region of the MMP-9 gene were distinguished by cutting with the SphI restriction enzyme. Genotype and allele frequencies were calculated, and data were analyzed by the chi2 test. There was a significant difference in MMP-9 genotypes between chronic periodontitis patients and healthy controls. The odds ratios for the CT genotype and the combination of CT and TT genotypes were 0.4 (95% confidence interval, 0.17 to 0.93; P=0.02) and 0.37 (95% confidence interval, 016 to 0.85; P=0.01) relative to the subjects with the CC genotype, respectively. MMP-9 promoter gene polymorphism seems to be associated with severe generalized chronic periodontitis.

Analysis of the MMP‐9 (C‐1562 T) and TIMP‐2 (G‐418C) gene promoter polymorphisms in patients with chronic periodontitis

Matrix metalloproteinases (MMP)-9 is an important member of the matrix metalloproteinase family. A functional polymorphism has been described in the promoter region of the human MMP-9 gene. A C-to-T base exchange at -1562 creates two different alleles, and the C/T and T/T genotypes promote high activity of the MMP-9 gene promoter, increasing the risk for inflammatory diseases. The metalloproteinase-2 tissue inhibitor (TIMP-2) regulates the activity of MMPs in the extracellular matrix, and a polymorphism at the -418 position of the TIMP-2 gene promoter has been found in a Sp-1 binding site. In this study we have investigated the association between the above-mentioned polymorphisms and chronic periodontitis severity. Genomic DNA from oral mucosa of 100 subjects was amplified by polymerase chain reaction and analysed by restriction endonuclease digestion. The significance of the differences in observed frequencies of polymorphisms in moderate and severe disease and healthy groups was assessed by chi(2) test (p<0.05). No association was observed between the polymorphism in the promoter region of MMP-9 (p=0.6693) and chronic periodontitis. The analysis of TIMP-2 showed that the G/G genotype was found at a frequency of 99%. The results show that the polymorphism in the promoter region of MMP-9 gene is not associated with chronic periodontitis. The high frequency of GG genotype in the TIMP-2 gene promoter in the population studied did not allow any conclusion regarding its effect on chronic periodontitis.

Shortening of microsatellite deoxy(CA) repeats involved in GL331-induced down-regulation of matrix metalloproteinase-9 gene expression

Matrix metalloproteinase-9 (MMP-9) associates with cancer cell invasion and metastasis. CL1-5 cells, a human lung adenocarcinoma cell line, expressed an elevated level of MMP-9 and exhibited a highly invasive and metastatic ability. By Matrigel assay and gelatinase zymography, the topoisomerase II poison GL331 was found to dose-dependently inhibit the invasiveness and the level of secreted MMP-9 of CL1-5 cells. Northern blot analysis indicated that cellular MMP-9 mRNA level was decreased after GL331 treatment. Furthermore, GL331-induced down-regulation of mmp-9 gene promoter was demonstrated by using a luciferase reporter gene driven by the −216 to −13 region of the mmp-9 gene promoter cloned from CL1-5 cells. By PCR amplification and gel electrophoresis, we found that GL331 caused shortening of the −216 to −13 region of the mmp-9 promoter. Direct sequencing analysis revealed that the number of d(CA) was reduced from 24 to 18 at the microsatellite d(CA) repeat region of the mmp-9 promoter. The CL1-5 cells transfected with the luciferase reporter containing 18 d(CA)s expressed only 53% of those when the reporter contained 24 d(CA)s. The promoter region of mmp-9 gene contains other positive regulatory elements, such as TRE and κB. We found that GL331 did not significantly influence the luciferase activity driven by TRE or κB. Taken together, these data suggested that GL331 inhibited MMP-9 mRNA expression at least partly through the selective induction of shortening of microsatellite d(CA) repeats. This is the first report that an anti-cancer agent can inhibit mmp-9 gene expression by inducing microsatellite DNA shortening.