SEARCH ASSOCIATION OF POLYMORPHISM rs3918242 IN MMP9 GENE WITH GUM RECESSION OF CHILDREN

Subject. Gingival recession is a complex multifactorial disease, the development of which is influenced by the interaction of many genes and environmental factors (orthodontic and mucogingival anomalies, bad habits, muscle hypertonicity). Since, in addition to environmental factors, genes also influence the development of this pathology, the study of the molecular genetic basis of gum recession is an urgent task of modern dentistry. Such studies can make a decisive contribution to the creation of a concept describing the development of gingival recession, will help determine the prospects for its personalization and will allow the development of scientifically based comprehensive programs for the prevention of periodontal pathology. The aim is to study the association of the rs3918242 of the metalloproteinase 9 (MMP9) gene in children with the development of gum recession citizensof the Republic of Tatarstan. Methodology. The study sample included 284 patients, of which 200 relatively healthy ones constituted the control group (intact periodontium) and 84 - the observation group (gingival recession). DNA was isolated from buccal epithelial cells. Further genotyping of the rs3918242 polymorphism of the MMP9gene was performed using real-time polymerase chain reaction on a CFX96 amplifier (BioRad, USA) in accordance with the manufacturer's instructions (Sib-DNA, Novosibirsk). Statistical analysis was performed to assess the differences between the study groups. Results. There were no statistically significant differences in the frequency of alleles and genotypes of the rs3918242 polymorphism of the MMP9 gene between the study groups (p > 0.05). Conclusions. The role of the rs3918242 polymorphism of the MMP9 gene in the development of gum recession in children living in the Republic of Tatarstan has not been established using the case-control methodological approach.

Association of the functionally significant polymorphisms of the MMP9 gene with H. pylori-positive gastric ulcer in the Caucasian population of Central Russia

Background and purpose The study analyzed the association of functionally significant polymorphisms of matrix metalloproteinases (MMPs) genes with the development of gastric ulcer (GU) in Caucasians from Central Russia. Methods The 781 participants, including 434 patients with GU (196 Helicobacter pylori (H. pylori)-positive and 238 H. pylori-negative) and 347 controls (all H. pylori-negative) were recruited for the study. Ten SNPs of the MMP1 (rs1799750), MMP2 (rs243865), MMP3 (rs679620), MMP8 (rs1940475), and MMP9 (rs3918242, rs3918249, rs3787268, rs17576, rs17577, and rs2250889) genes were considered for association with GU using multiple logistic regression. The SNPs associated with GU and loci linked (r2≥0.8) to them were analyzed in silico for their functional assignments. Results The SNPs of the MMP9 gene were associated with H. pylori-positive GU: alleles C of rs3918249 (OR = 2.02, pperm = 0.008) and A of rs3787268 (OR = 1.60–1.82, pperm ≤ 0.016), and eight haplotypes of all studied MMP9 gene SNPs (OR = 1.85–2.04, pperm ≤ 0.016) increased risk for H. pylori-positive GU. None of the analyzed SNPs was independently associated with GU and H. pylori-negative GU. Two haplotypes of the MMP9 gene (contributed by rs3918242, rs3918249, rs17576, and rs3787268) increased risk for GU (OR = 1.62–1.65, pperm ≤ 0.006). Six loci of the MMP9 gene, which are associated with H. pylori-positive GU, and 65 SNPs linked to them manifest significant epigenetic effects, have pronounced eQTL (17 genes) and sQTL (6 genes) values. Conclusion SNPs of the MMP9 were associated with H. pylori-positive GU but not with H. pylori-negative GU in Caucasians of Central Russia.

The Effect of Neddylation Inhibition on Inflammation-Induced MMP9 Gene Expression in Esophageal Squamous Cell Carcinoma

Inhibition of the protein neddylation process by the small-molecule inhibitor MLN4924 has been recently indicated as a promising direction for cancer treatment. However, the knowledge of all biological consequences of MLN4924 for cancer cells is still incomplete. Here, we report that MLN4924 inhibits tumor necrosis factor-alpha (TNF-α)-induced matrix metalloproteinase 9 (MMP9)-driven cell migration. Using real-time polymerase chain reaction (PCR) and gelatin zymography, we found that MLN4924 inhibited expression and activity of MMP9 at the messenger RNA (mRNA) and protein levels in both resting cells and cells stimulated with TNF-α, and this inhibition was closely related to impaired cell migration. We also revealed that MLN4924, similar to TNF-α, induced phosphorylation of inhibitor of nuclear factor kappa B-alpha (IκB-α). However, contrary to TNF-α, MLN4924 did not induce IκB-α degradation in treated cells. In coimmunoprecipitation experiments, nuclear IκB-α which formed complexes with nuclear factor kappa B p65 subunit (NFκB/p65) was found to be highly phosphorylated at Ser32 in the cells treated with MLN4924, but not in the cells treated with TNF-α alone. Moreover, in the presence of MLN4924, nuclear NFκB/p65 complexes were found to be enriched in c-Jun and cyclin dependent kinase inhibitor 1 A (CDKN1A/p21) proteins. In these cells, NFκB/p65 was unable to bind to the MMP9 gene promoter, which was confirmed by the chromatin immunoprecipitation (ChIP) assay. Taken together, our findings identified MLN4924 as a suppressor of TNF-α-induced MMP9-driven cell migration in esophageal squamous cell carcinoma (ESCC), likely acting by affecting the nuclear ubiquitin–proteasome system that governs NFκB/p65 complex formation and its DNA binding activity in regard to the MMP9 promoter, suggesting that inhibition of neddylation might be a new therapeutic strategy to prevent invasion/metastasis in ESCC patients.

Relationship between rs3918242 Polymorphism of Matrix Methaloproteinase-9 Gene and Preeclampsia in Pregnant Women

Background: Preeclampsia is a complex disorder of pregnancy with an unknown etiology. Numerous studies have shown the possible role of gene polymorphisms, especially metalloproteinases, in development of this disease, but there are no definitive results. Objective: This study aims to investigate the possible association between rs3918242 (−1562C>T) polymorphism in Matrix Metalloproteinase 9 (MMP9) gene with the risk of preeclampsia in pregnant women. Methods: In this cross-sectional study, participants were 90 pregnant women with preeclampsia and 199 healthy pregnant women (controls). The genotypes of rs3918242 polymorphism were investigated using Polymerase Chain Reaction technique and Limited Fragment Length Polymorphism method. Logistic regression analysis was used to investigate the relationship between rs3918242 polymorphism and preeclampsia. Findings: The frequency of CC, CT, TT genotypes of rs3918242 polymorphism was reported 47.8%, 47.8% and 4.2% in patients and 84.8, 13.1 and 2% in controls, respectively, and the difference between groups was significant (P<0.001). The frequency of TT genotype in patients was significantly higher than in controls (P<0.001). Moreover, the frequency of T allele in patients was 52.2%, while in controls it was 15.2% and the difference between the two groups was significant (P<0.001). Conclusion: The rs3918242 polymorphism of MMP9 gene plays an important role in the incidence of preeclampsia in pregnant women.

The Effect of TNF-α on the Expression of MMP9 in Human Mesenchymal Bone Marrow-Derived Stem Cells

Matrix metalloproteinase 9 (MMP9) as the enzyme of adult stem cells secreted from damage cells. In spite of low level of MMP9 enzyme in the mesenchymal stem cells, many inflammatory cytokines stimulation such as TNF-&alpha; could increase MMP9 level in cells. Current study evaluated the expression of the MMP9 enzyme under the influence of TNF-&alpha; in human bone marrow mesenchymal stem cells. The human bone marrow mesenchymal stem cells were classified into control and experimental groups. In the experimental groups, various concentrations of the TNF-&alpha; (1ng/ml and 10ng/ml) were administrated in different times (10 and 24 hours), whereas the control group was not treated with TNF-&alpha;. MMP9 gene expression was evaluated by Real-Time PCR. TNF-&alpha; administration in 1ng/ml and 10ng/ml dosage for 10 hours, induced the expression of MMP9 1468.3 and 1782.8 times more than the control group, respectively. After 24h, in comparison between 1ng/ml and 10ng/ml with control groups, MMP9 expression were 442.64 and 1184.4 times more than control group, respectively. In conclusion, the expression rate of the MMP9 gene in bone marrow mesenchymal stem cells might be effected by dosage and time of exposure to TNF-&alpha;. Furthermore, the time of exposure might have the prominent role in alteration of MMP9 gene expression induction in the mesenchymal stem cells.

Effects of Toxicity Induced by Gentamicin on the Kidney of Killifish Aphaniops hormuzensis and the Role of Wt1 and MMP9 Genes in Response to This Toxicity

Background: Aminoglycoside antibiotics such as gentamicin are used to cure bacterial infections in humans and other animals, but they can cause nephritic damage, as well. Nephrotoxicity is one of the side effects of gentamicin. Objectives: The objective of this study was to investigate the effects of toxicity induced by gentamicin on the kidney of killifish Aphaniops hormuzensis. Also, we aimed to study the expression pattern of Wt1 and MMP9 genes by real-time PCR in response to this toxicity. Methods: First, 10 µg/g (sub-lethal dose) gentamicin was given to adult fish. The kidney tissues were dissected and preserved in 10% formalin for a 24-hour; then, they underwent standard histological procedures. The sections were prepared at 3 μm and stained with Haematoxylin & Eosin (H&E). The slide microphotography process was done by an Olympus CH2 microscope. The RNA was isolated, and cDNA was synthesized with a standard protocol, and the expression patterns of Wt1 and MMP9 genes were examined by real-time PCR. Results: Nephrotoxicity occurred 10 hours after the injection of gentamicin, and the injury was detected in the epithelium of kidney tubules. The kidney tubule regenerated itself within 10 days post-injection (dpi). On 7 dpi, the nephrogenic body formation occurred and was differentiated into renal nephrons. The Wt1 gene was upregulated (two-fold) on 5 dpi after kidney damage and then had a down-regulation on 7 dpi when the kidney began to regenerate. The MMP9 gene showed increased expression in comparison with the control sample in the study days, and this expression increased on 7 dpi by 6.6 folds. Conclusions: The results of this study, for the first time, highlighted that nephritic damage appears in the kidney of A. hormuzensis after toxicity induced by gentamicin and that changes in the expression of the examined genes are consistent with their roles in the process of renal regeneration in this species.

Association of a combination of alleles and genotypes of polymorphic variants of matrix metalloproteinase genes and their tissue inhibitors genes with a risk of developing of gastric cancer

Проведен анализ распределения частот аллелей/генотипов полиморфных локусов rs1799750 и rs494379 гена MMP1, rs2285053 гена MMP2, rs3025058 гена MMP3, rs3918242 и rs17576 гена MMP9, rs2276109 гена MMP12, rs8179090 гена TIMP2 и rs9619311 гена TIMP3 у 314 пациентов с раком желудка, а также у 339 здоровых индивидов из Республики Башкортостан. С помощью алгоритма APSampler выявлены сочетания аллелей/генотипов, ассоциированные как с повышенным, так и с пониженным риском развития заболевания. The analysis of the frequency distribution of alleles/genotypes of polymorphic loci rs1799750 and rs494379 of the MMP1 gene, rs2285053 of the MMP2 gene, rs3025058 of the MMP3 gene, rs3918242 and rs17576 of the MMP9 gene, rs2276109 of the MMP12 gene, rs8179090 of the TIMP2 gene and rs9619311 of the TIMP3 gene in 314 patients with gastric cancer, as well as in 339 healthy individuals from the Republic of Bashkortostan. Using the APSampler algorithm were identified combinations of alleles/genotypes associated with increased and reduced risk of developing the disease.

Association Study between Functional Polymorphisms of MMP9 Gene Promoter and Multiple Sclerosis Susceptibility in an Iranian Population

Background: Matrix metalloproteinase-9 (MMP-9) polymorphisms, C−1562 T and -90 (CA) n repeats, which influence transcriptional activity of this gene, are proposed to play a role in MS susceptibility and its development. In the present study, the possible association of MMP-9 polymorphisms in Iranian MS patients is studied. Methods: Association of MMP-9 mentioned gene polymorphisms with MS susceptibility was evaluated in unrelated Iranian subjects referred to Al-Zahra Hospital, Isfahan, Iran during 2014 to 2017. Results: -1562 T allele of MMP-9 was associated with increased MS risk. However, we found no overall significant effect of −90 (CA)n repeat on MS susceptibility. Conclusion: For as much as MMP-9 molecule is a potential target for MS therapy, to determine whether any of MMP-9 polymorphisms influence MS susceptibility in Iranian MS patients or not, concerning the significant influence of T allele on MS susceptibility and the non-significant association regarding CA repeats, further research is needed before proposing any definite conclusion.