Matrix Metalloproteinase-9 Expression Research Articles

LncRNA NEAT1 promotes proliferation of ovarian cancer cells and angiogenesis of co-incubated human umbilical vein endothelial cells by regulating FGF9 through sponging miR-365: An experimental study.

Objective:To uncover the function of lncRNA NEAT1 in ovarian cancer (OC) cells and its mechanism.Methods:The expression patterns of lncRNA NEAT1 and FGF9 in human OC cells and human ovarian epithelial cells was determined. OC cells were transfected with sh-NEAT1, pcDNA3.1-NEAT1, miR-365 mimic, miR-365 inhibitor or pcDNA3.1-NEAT1 + sh-NEAT1 before cell proliferation rate and cell clone formation rate were measured. After the transfected OC cells were co-cultivated with human umbilical vein endothelial cells (HUVECs), Matrigel angiogenesis assay tested angiogenesis of HUVECs; qRT-PCR and Western blot tested the expressions of vascular endothelial growth factor (VEGF), angiogenin 1 (Ang-1) and matrix metalloproteinase 2 (MMP2). Dual-luciferase reporter assay determined the targeted binding of NEAT1 and FGF9 to miR-365.Results:LncRNA NEAT1 and FGF9 are over-expressed in OC cells. Knockdown of NEAT1 or FGF9, or over-expression of miR-365 results in decreased proliferation rate and cell clones as well as inhibited angiogenesis and down-regulated expressions of VEGF, Ang-1 and MMP2. Over-expression of NEAT1 or knockdown of miR-365 can reverse the effect caused by FGF9 knockdown. NEAT1 can down-regulate the expression of miR-365 while up-regulating that of FGF9. Dual-luciferase reporter assay determined that NEAT1 competes with FGF9 for binding to miR-365.Conclusion:LncRNA NEAT1 up-regulates FGF9 by sponging miR-365, thus promoting OC cell proliferation and angiogenesis of HUVECs.

Matrix Metalloproteinase-9 Expression is Enhanced by Ischemia and Tissue Plasminogen Activator and Induces Hemorrhage, Disability and Mortality in Experimental Stroke

Matrix metalloproteinase-9 (MMP-9) degrades collagen and other cellular matrix proteins. After acute ischemic stroke, increased MMP-9 levels are correlated with hemorrhage, lack of reperfusion and stroke severity. Nevertheless, definitive data that MMP-9 itself causes poor outcomes in ischemic stroke are limited. In a model of experimental ischemic stroke with reperfusion, we examined whether ischemia and recombinant tissue plasminogen activator (r-tPA) therapy affected MMP-9 expression, and we used specific inhibitors to test if MMP-9 affects brain injury and recovery. After stroke, MMP-9 expression increased significantly in the ischemic vs. non-ischemic hemisphere of the brain (p < 0.001). MMP-9 expression in the ischemic, but not the non-ischemic hemisphere, was further increased by r-tPA treatment (p < 0.001). To determine whether MMP-9 expression contributed to stroke outcomes after r-tPA treatment, we tested three different antibody MMP-9 inhibitors. When compared to treatment with r-tPA and saline, treatment with r-tPA and MMP-9 antibody inhibitors significantly reduced brain hemorrhage by 11.3 to 38.6-fold (p < 0.01), brain swelling by 2.8 to 4.3-fold (p < 0.001) and brain infarction by 2.5 to 3.9-fold (p < 0.0001). Similarly, when compared to treatment with r-tPA and saline, treatment with r-tPA and an MMP-9 antibody inhibitor significantly improved neurobehavioral outcomes (p < 0.001), decreased weight loss (p < 0.001) and prolonged survival (p < 0.01). In summary, both prolonged ischemia and r-tPA selectively enhanced MMP-9 expression in the ischemic hemisphere. When administered with r-tPA, specific MMP-9 inhibitors markedly reduced brain hemorrhage, swelling, infarction, disability and death, which suggests that blocking the deleterious effects of MMP-9 may improve outcomes after ischemic stroke.

Evaluation of MMP-9 and iNOS expressions in sheep with encephalitic listeriosis

This study aimed to correlate Matrix Metalloproteinase-9 and iNOS expressions with the severity of histopathological findings in tissue samples taken from sheep with encephalitic listeriosis. Thus, the role of these molecules in the pathogenesis of the disease can be elucidated. After systemic necropsy, tissue samples of adult sheeps with meningoencephalitis were investigated by the culture, histopathological and immunohistochemical methods in the presence of Listeria spp. isolation from tissues was performed in accordance with the USDA-FSIS method with some modifications. Tissue samples were fixed in a 10% buffered formaldehyde solution. Following routine procedures, tissue sections at 5 μm were stained with Hematoxylin and Eosin, investigated under light microscope and photographed. Immunohistochemical staining was performed on the tissues using the avidin-biotin immune peroxidase complex method. Listeria spp. were obtained in 20 (83.3%) of 24 tissue samples with the presence of bright grey-black centred smooth colonies on Listeria Selective Agar and identified as Listeria monocytogenes through the phenotypically supportive tests. Liquefaction necrosis, purulent meningoencephalitis, perivascular cuffing, microabscesses and glial nodules were the most important histopathological findings. MMP-9 immunpositive reactions were observed in the cytoplasm of microglial cells and neurons in areas where inflammatory and necrotic areas are concentrated in medulla oblongata and pons. In perivascular cuffing areas, immune reactions in endothelial cells were detected. We detected iNOS positive reactions in the medulla oblangata and pons, especially in inflammatory cells in the microabscesses. Consequently, a positive correlation (p &amp;lt; 0.05) was found between MMP-9 expression and the severity of histopathological findings in sheep with encephalitic listeriosis. expression. In addition, we found that iNOS expression increased in parallel with the increase in MMP-9

Fenofibrate attenuates doxorubicin-induced cardiac dysfunction in mice via activating the eNOS/EPC pathway

Endothelial progenitor cells (EPCs) improve endothelial impairment, which in turn restores endothelial function in patients with heart failure (HF). In the present study, we tested whether fenofibrate, with its anti-inflammatory and vasoprotective effects, could improve myocardial function by activating EPCs through the eNOS pathway in a doxorubicin (DOX)-induced cardiomyopathy mouse model. Wild-type mice were divided into 4 groups and treated with vehicle, DOX + saline, DOX + fenofibrate, and DOX + fenofibrate + L-NAME (N(ω)-nitro-L-arginine methyl ester). DOX-induced cardiac atrophy, myocardial dysfunction, the number of circulating EPCs and tissue inflammation were analyzed. Mice in the DOX + fenofibrate group had more circulating EPCs than those in the DOX + saline group (2% versus 0.5% of total events, respectively) after 4 weeks of treatment with fenofibrate. In addition, the inhibition of eNOS by L-NAME in vivo further abolished the fenofibrate-induced suppression of DOX-induced cardiotoxic effects. Protein assays revealed that, after DOX treatment, the differential expression of MMP-2 (matrix metalloproteinase-2), MMP-9 (matrix metalloproteinase-9), TNF-α (tumor necrosis factor-α), and NT-pro-BNP (N-terminal pro-B-type natriuretic peptide) between saline- and DOX-treated mice was involved in the progression of HF. Mechanistically, fenofibrate promotes Akt/eNOS and VEGF (vascular endothelial growth factor), which results in the activation of EPC pathways, thereby ameliorating DOX-induced cardiac toxicity.

The Effect and Mechanism of Asymmetric Dimethylarginine Regulating Trophoblastic Autophagy on Fetal Growth Restriction.

Fetal growth restriction (FGR) is an important cause of perinatal death and adverse pregnancy outcomes. Asymmetric dimethylarginine (ADMA) is associated with FGR, but the mechanisms have not been thoroughly studied. Here, we determined the levels of ADMA and autophagy-related molecules in human blood samples and placental tissues. And we also used the human chorionic carcinoma cell line BeWo to investigate the mechanism of ADMA-induced FGR in vitro. Compared with the control group, ADMA levels in maternal blood and placenta were increased in patients with FGR, and the birth weight (BW) percentile was negatively correlated with maternal serum ADMA concentration in the FGR group. The expression of mammalian target of rapamycin (mTOR) in the placenta of the FGR group was lower than the control group, while the expression of Beclin-1 and microtubule-associated protein 1 light chain 3-II (LC3-II)/LC3-I was significantly increased in the FGR group. And the expression of matrix metalloproteinase 9 (MMP9) was decreased in the placenta of patients with FGR. In in vitro cell experiments, compared with the control group, the expression of mTOR and MMP9 in BeWo cells was decreased and the expression of Beclin-1 and LC3-II/LC3-I was increased in the ADMA-treated group. Moreover, ADMA had favorable effects on the formation of autophagic vacuoles, and the autophagy inhibitor 3-Methyladenine (3-MA) could reduce the autophagy-induction effect of ADMA on BeWo cells. This study found that ADMA could participate in the occurrence of FGR through inducing autophagy in trophoblasts.

Correlation of Matrix Metalloproteinase-2 and p21 Expressions with Capsular Invasion of Thymoma AB

Thymomas are the common neoplasms of mediastinum, and type AB is the most frequent case. All thymomasare potential for invasion to adjacent tissues and considered malignant. Capsular invasion is one of itsprognostic indicators. Matrix metalloproteinase-2 (MMP-2) is a family of proteinases that has the abilityto degrade a component of extracellular matrix and affect tumor invasion and or metastasis. Cytoplasmicexpression of p21 protein can be significantly correlated with invasion and metastasis. The correlation of thesetwo proteins in thymoma has not been widely studied, hence we aimed to analyze the expression of MMP-2and p21 and investigate their correlation with capsular invasion of thymoma AB. This cross-sectional studywas performed on the 24 paraffin-embedded samples of thymomectomy during January 2013-Desember2019 at Anatomical Pathology Laboratory of Dr. Soetomo General Hospital Surabaya. The samples weredevided based on capsular invasion into 2 groups. Immunohistochemical staining was performed to detectexpression of p21 and MMP-2. The correlation was statistically analyzed using Spearman test. There wasno significant difference of MMP-2 expression between thymoma with capsular invasion and withoutcapsular invasion (p=0.839), and also no significant difference of p21 expression between thymoma withcapsular invasion and without capsular invasion (p=0.816). No correlation of MMP-2 and p21 expressionsin thymoma AB was revealed (p=0.255). In thymoma AB, the expression of MMP-2 and p21 were notcorrelate with capsular invasion. These results may contribute to the development of thymoma research.

PECULIARITIES OF BRAIN-DERIVED NEUROTROPHIC FACTOR AND MATRIX METALLOPROTEINASE-9 EXPRESSION DYNAMICS IN PATIENTS WITH PARANOID SCHIZOPHRENIA DEPENDING ON THE DURATION OF THE DISEASE

The aim: To study the expression of brain-derived neurotrophic factor (BDNF) and matrix metalloproteinase-9 (MMP-9) in the blood serum of patients with paranoid schizophrenia and to trace the features of their dynamics depending on the duration of the disease and analyse the correlation between BDNF, MMP-9 serum levels and symptoms severity by using the Positive and Negative Syndrome Scale (PANSS). Materials and methods: The study included 120 patients, namely 20 patients with paranoid schizophrenia diagnosed less than 3 years ago (Сomparison Group) and 100 patients with a diagnosis of paranoid schizophrenia (Study Group): 20 of them have been suffering from this disease from 3 to 5 years (Subgroup I); 10 patients – from 5 to 10 years (Subgroup II); 10 patients – from 10 to 15 years (Subgroup III); 10 patients – from 15 to 20 years (Subgroup IV); 10 patients – from 25 years and more (Subgroup V). The groups did not differ with respect to age or gender. The content of BDNF and MMP-9 in blood serum was determined by enzyme-linked immunosorbent assay. Results: BDNF concentration averaged 28.327 ± 5.32 pg/ml in the patients of Group I; 25.40 ± 2.31 pg/ml in Group II; 24.32 ± 3.1 pg/ml in Group III; 23.8 ± 1.32 pg/ml in Group IV; 21.39 ± 0.97 pg/ml in Group V; 9.36 ± 4.38 pg/ml in Group VI. The expression of MMP-9 in the experimental groups constituted: 942.84 ± 87.80 pg/ml, 1042.84 ± 87.80 pg/ml, 1142.53 ± 77.20 pg/ml, 1752.84 ± 77.80 pg/ml, 1542.84 ± 37.70 pg/ml, 2042.74 ± 47.80 pg/ml, respectively. Decreased BDNF negatively correlated with MMP-9 expression (r=0.46; p&lt;0.05). Conclusions: The development of paranoid schizophrenia was manifested by an imbalance in BDNF level and MMP-9 expression which could affect neurogenesis, synapticplasticity, ability to learn and remember, therefore, they could be considered as diagnostic markers of the pathology. With the increase in the duration of the studied pathology, BDNF parameters decreased and MMP-9 expression increased. A negative correlation between them was noted.

Effect of melanocyte keratinocyte transplantation procedure, using ablative CO2 laser and followed by NB-UVB phototherapy, on the expression of E-Cadherin and matrix metalloproteinase-2 in acral vitiligo: an immunohistochemical study

Background Melanocyte keratinocyte transplantation procedure (MKTP) is a multi-procedural intervention that could hypothetically alter the environment surrounding grafted epidermal cells, helping melanocytes’ migration and adherence to keratinocytes in their basal position. Objective To evaluate the effect of MKTP in acral vitiligo skin prepared by total ablative CO2 laser resurfacing and followed by narrow-band ultraviolet B (NB-UVB) phototherapy on matrix metalloproteinase-2 (MMP2) and E-Cadherin expression and the reflection of these changes on repigmentation. Patients and methods Acral lesions in 20 stable nonsegmental vitiligo patients were prepared by full CO2 ablation down to the papillary dermis followed by MKTP and NB-UVB sessions. Two 4 mm punch biopsies were taken: the first at baseline and the second after the onset of repigmentation or after 6 months of phototherapy if no repigmentation occurred. Immunohistochemical staining for evaluation of E-Cadherin and MMP2 expression was performed. Results Eight patients (40%) achieved repigmentation ranging from 10 to 90% with a median of 62.5%. E-Cadherin intensity was significantly increased after treatment (P Conclusion MKTP with recipient site prepared by full CO2 laser ablation followed by NB-UVB phototherapy increased the expression of E-Cadherin and MMP2 in acral vitiliginous skin. The increase of dermal MMP2 could play a favorable role in repigmentation.

Assessment of the correlation between oxidative stress and expression of MMP-2, TIMP-1 and COX-2 in human aortic smooth muscle cells.

IntroductionSmooth muscle cells (SMCs) are considered to be the main producer of matrix metalloproteinase-2 (MMP-2) participating primarily in extracellular matrix (ECM) remodeling. Any disturbances in ECM structure may underlie the pathogenesis of many cardiovascular diseases and contribute to angiogenesis, cancer development, invasion or metastasis. The purpose of the study was to examine the effect of oxidative stress on the expression of MMP-2, its tissue inhibitor type 1 (TIMP-1) and cyclooxygenase-2 (COX-2) in human aortic smooth muscle cells (HASMCs).Material and methodsHASMCs were treated with exogenously applied H2O2 or TNF-α. N-acetylcysteine (NAC) was used as an antioxidant. Gene expression levels were measured by real-time PCR and the protein levels were determined using ELISA assay.ResultsThe studies revealed no association between oxidative stress and either mRNA quantity or protein secretion of MMP-2 and TIMP-1. However, we found markedly reduced (p < 0.001) MMP-2 secretion in cells incubated with NAC. HASMCs stimulated with TNF-α demonstrated a significantly increased COX-2 mRNA level as well as enzyme activity. H2O2-induced cells showed lowered COX-2 activity in comparison to untreated cells. MMP-2 and TIMP-1 expression did not change after COX-2 inhibition with DuP-697.ConclusionsWe did not find any effect of oxidative stress on expression of MMP-2 and TIMP-1 in HASMCs. However, COX-2 mRNA and protein level were elevated in these conditions. There was no correlation between COX-2 activity and MMP-2 and TIMP-1 mRNA expression or protein secretion.

Expression of tumor necrosis factor Alpha, interleukin-1 and matrix metalloproteinase-9 and pathomorphological changes in acquired middle ear cholesteatoma

&amp;lt;p&amp;gt;Introduction. The inflammatory mediators play a central role in the pathogenesis of the inflammatory process of the middle ear and cholesteatoma from the aspect of initiating and maintaining the inflammatory response to infection and lesion. The aim of the study was to examine if the presence of acquired cholesteatoma could predict pathomorphological changes of the tympanic cavity mucosa in relation to the control tissue of the inflamed middle ear mucosa and to examine and compare the expression levels of tumor necrosis factor-alpha (TNF-a), interleukin-1 (IL-1) and matrix metalloproteinase 9 (MMP-9) with pathomorphological changes in the middle ear mucosa in chronic otitis media (COM), with and without acquired cholesteatoma (AC). Methods. The immunohistochemical study included 178 patients of both sexes, aged 5 to 75, who underwent microsurgical treatment of COM from 2015 to 2018. Patients were divided into two groups based on the presence or absence of AC of the middle ear: 97 with cholesteatoma (CCOM) and 81 without cholesteatoma (COM). Samples of the perimatrix of AC and inflamed middle ear mucosa were taken intraoperatively. The condition of the tympanic cavity mucosa was examined by otomicroscopy exploration intraoperatively. The expression levels of TNF-a, IL-1 and MMP-9 were determined by immunohistochemical analysis. Results. The difference in the percentage distribution of patients according to the condition of the tympanic cavity mucosa between both groups was statistically significant (p &amp;amp;lt;0.01) where in the COM group the highest frequency was 43.2% of patients with mucosal hypertrophy, and in the CCOM 56.7% with granulations. With highly positive expression of TNF-R2 and IL-1, a higher probability of the presence of mucosal hypertrophy and granulations can be expected, and with highly positive expression of MMP-9 the presence of granulations. Conclusion. Acquired middle ear cholesteatoma is a statistically significant predictor of the occurence of mucosal hypertrophy and granulations in the tympanic cavity in relation to the control tissue of the inflamed middle ear mucosa. The high expression of TNF-R2, IL-1 and MMP-9 shows a statistically significant association with the presence of granulations and mucosal hypertrophy in acquired middle ear cholesteatoma which may have clinical significance in the evaluation and prognosis of the disease.&amp;lt;/p&amp;gt;

Matrix Metalloproteinase-2 Expression as a Prognostic Marker in Breast Carcinomas

Introduction: Matrix Metalloproteinase-2 (MMP-2) is over expressed in a variety of malignant tumours and their expression and activity are often associated with tumour aggressiveness and a poor prognosis. It serves as a prognostic marker in breast carcinoma regardless of patient age, disease stage, malignancy grade, or hormone receptor status and modulation of MMP-2 expression and activation provides a new mechanism for breast cancer treatment. Aim: To evaluate the expression of MMP-2 in breast carcinoma tumour cells and peritumoural stroma and also to analyse the findings with the existing other prognostic markers of mammary carcinoma. Materials and Methods: The present study was a retrospective study conducted on paraffin blocks of 90 cases of invasive breast carcinoma specimens received in the Department of Pathology, Sri Raamachandra Institute of Higher Education and Research a tertiary care centre in Chennai, Tamil Nadu, India, from January 2012 to June 2017. Immunohistochemical staining for MMP-2, Oestrogen Receptor (ER), Progesterone Receptor (PR) and Human Epidermal Growth Factor Receptor 2 (HER2) was done. Statistical analysis was done on the data collected by using the software GNU-PSPP version 0.10.1. Pearson Chi-square test was used to determine significant clinicopathological differences between MMP-2 expression in positive and negative tumours. Differences were considered statistically significant when p-value was &lt;0.05. Results: The study included 90 cases of histological proven invasive breast carcinoma. The study parameters include age, clinical staging, histopathological grade, lymphnode status, molecular subtype and MMP-2 expression in invasive breast carcinoma. Out of 90 cases, 62 cases were positive for MMP-2 and 18 cases were positive for peritumoural stroma, 30 cases were negative for MMP-2 in tumoural cells and 72 cases were negative surrounding the peritumoural stroma. Conclusion:Present study showed high MMP-2 immunohistochemical expression in breast carcinomas. There was a statistically significant association of increased MMP-2 expression with tumour stage. There was also a statistically significant association of MMP-2 in the tumour and stroma with High Grade Ductal Carcinoma In Situ (HGDCIS). There was an increased expression of MMP-2 in Luminal A subtype. Low expression was seen in the other molecular subtypes. In view of these findings and association with other studies in the literature, the present study demonstrates that the expression of MMP-2 in tumour and stromal cells could serve as a parameter of poor prognosis in breast cancer.

Expression of Matrix Metalloproteinase-9 in Different Histopathological Variants of Ameloblastoma: A Cross-sectional Study

Introduction: Ameloblastomas are benign and the most common odontogenic neoplasms with many histopathological subtypes depending on the predominant pattern. They are known for their aggressive behaviour. As ameloblastomas have a high rate of recurrence, it is necessary to understand the biological behaviour of these neoplasms. Matrix Metalloproteinase-9 (MMP-9) is an enzyme that belongs to Metalloproteinases family and is known to degrade the Extracellular Matrix (ECM) and facilitate tumour progression. Evaluation of the expression of MMP-9 in ameloblastomas could contribute in understanding its biological behaviour. Aim: To analyse the expression of MMP-9 in different histopathological variants of ameloblastoma. Materials and Methods: A cross-sectional observational study was done in the Department of Oral Pathology and Microbiology, Bharati Vidyapeeth Deemed to be University, Dental College and Hospital, Pune, India. A total of 30 cases were selected for this study. The study was carried out for a duration of two years from April 2018 to April 2020. The MMP-9 expression was studied by immunohistochemical staining. The statistical comparison was done using Chi-square test between groups, p-value &lt;0.05 was considered significant. The data was statistically analysed using Statistical Package for Social Sciences (SPSS) version 22.0, IBM Corporation, USA for MS Windows. Results: A total of 30 paraffin embedded archival tissue blocks were selected for this study. Among them nine cases were of Plexiform Ameloblastoma, eight of Unicystic Ameloblastoma, three of Acanthomatous Ameloblastoma, four of Desmoplastic Ameloblastoma and six were of Follicular Ameloblastomas. The MMP-9 showed variable expression in different histopathological subtypes of ameloblastoma. This difference was statistically significant between Plexiform and Acanthomatous as well as Plexiform and Follicular variants (p&lt;0.05). A 66.7% (6 out of 9 samples) of Plexiform Ameloblastoma showed intense staining for MMP-9. Conclusion: Expression of MMP-9 varies in different histopathological variants of ameloblastoma and may not have an association with biological behaviour and aggressiveness.

Dynamic change of MMP-9 in diabetic stroke visualized by optical imaging and treated with CD28 superagonist.

Type 2 diabetes mellitus is associated with an increased risk for stroke and unfavorable outcomes following stroke. Matrix metalloproteinase-9 (MMP-9) is a potential contributor to the poor prognosis of diabetic ischemic stroke. Investigations on diabetic stroke are limited by the lack of non-invasive imaging techniques. In this study, we report a fast and ultra-sensitive MMP-activatable optical imaging probe (MMP-P12) to achieve non-invasive and real-time visualization of the dynamic expression of MMP-9 in diabetic stroke. Moreover, by using this probe, we aim to detect the therapeutic efficacy of CD28 SA in diabetic stroke. Serial near-infrared fluorescence (NIRF) imaging was performed on wild-type and STZ-induced diabetic mice after MMP-P12 probe injection on days 1, 3, and 7 post ischemic stroke. The dynamic change in MMP-9 expression after CD28 SA treatment was also imaged on days 1, 3, and 7 post stroke and confirmed by immunohistochemistry staining and western blotting. NIRF imaging showed that diabetic stroke mice presented a trend of higher levels of MMP-9. CD28 SA treatment significantly downregulated the expression of MMP-9 on day 7 post stroke. Glucose also had a downward trend in CD28 SA treated diabetic stroke mice. In conclusion, our data suggest that MMP-P12 probe successfully detect the dynamic change of MMP-9 in diabetic stroke by utilizing optical imaging.CD28 SA treatment decreased the expression of MMP-9 and could be a promising therapeutic strategy for the treatment of diabetic stroke.

Epidermal Growth Factor Is Increased in Conjunctival Malignant Melanoma.

Conjunctival malignant melanoma (CMM) is a rare, but very aggressive tumor with a high metastasis rate. Not much is known about the CMM metastasis mechanisms. So far, epidermal growth factor (EGF) and its receptor (EGF-R) as well as macrophages and matrix metalloproteinase 9 (MMP-9) have been reported to lead to metastasis by epithelial-mesenchymal-transition and tumor migration in different solid tumors. Therefore, we evaluated whether EGF and EGF-R, CD68 and MMP-9 are altered in CMM samples in comparison to conjunctival nevi and healthy conjunctiva. EGF, EGF-R, the macrophage marker CD68 and MMP-9 expression were analyzed in human conjunctival melanoma (CMM, n=16), human conjunctival nevi (n=13) and disease-free human conjunctiva (controls, n=14) by immunohistology. Staining of each sample was evaluated using a standardized score ranging from negative (0) to triple positive (3). The groups were then compared by ANOVA, followed by Tukey's post-hoc test. A statistically significant increase of EGF was seen in CMM samples in comparison to conjunctival nevi (p=0.03). In contrast, no statistically significant differences in EGF-R expression were noted between the three groups. A statistically significant increase of CD68 was only seen in conjunctival nevi compared to controls (p=0.04). MMP-9 expression was similar in all groups. In CMM, the study data demonstrated an up-regulation of EGF in comparison to conjunctival nevi. Hence, EGF might promote proliferation of CMM cells and induce the epithelial-mesenchymal transition. Therefore, our data suggest that an interplay between EGF and CMM might have a critical role in the developing CMM tumors and metastasis.

Correlation Between the Expression of Matrix Metalloproteinase-9, Matrix Metalloproteinase-13, Tissue Inhibitor of Metalloproteinases-1, p16 and Differentiation of Head and Neck Squamous Cell Carcinoma: A Prospective Observational Study.

Introduction:The expression of matrix metalloproteinase-9 (MMP-9), MMP-13, and tissue inhibitor of metalloproteinases (TIMP-1) in head and neck squamous cell carcinoma (HNSCC) could be a useful predictor of tumour differentiation, nodal metastasis, and invasiveness. We conducted this study to ascertain the correlation between the expression of these markers and differentiation of tumour cells.Materials and Methods:A prospective observational study was conducted in a tertiary care center. Forty-three cases of proven HNSCC were recruited after obtaining informed consent. Using the surgically excised specimen, tumour differentiation and invasiveness were assessed and correlated with rates of expression of the markers. Chi-square test was done to correlate immunohistochemical (IHC) marker positivity and the degree of differentiation of the tumour, lymph node metastasis, and invasiveness.Results:MMP-9, MMP-13, and TIMP-1 were expressed in 72%, 34%, and 18% of cases, respectively. p16 expression was not found in any of the cases. MMP-13 expression correlated with poorer differentiation of the tumour (p = 0.03), and relatively younger age at diagnosis (p = 0.01). However, there was no correlation with lymphovascular or perineural invasion or lymph node metastasis.Discussion:In our study, MMP-13 expression correlated with poorer tumour differentiation and younger age at diagnosis, giving indirect evidence of tumour aggressiveness. IHC markers can provide additional information to prognosticate HNSCC. Identifying potential targets for newer biological therapy is essential in the Indian population as there are biological differences in cancer behavior. Increased expression of the proteolytic MMP-13 correlated with poorer differentiation of HNSCC.

The Activation of ROS/NF-κB/MMP-9 Pathway Promotes Calcium-Induced Kidney Crystal Deposition.

Idiopathic hypercalciuria is an important risk factor for the formation of calcium-containing kidney stones. Matrix metalloproteinase-9 (MMP-9) is closely related to cell and tissue remodeling and is involved in ectopic tissue calcification. However, little is known about its role in kidney stone formation. In this study, we found that the expression of MMP-9 and that of osteoblastic-related proteins was increased in normal rat kidney epithelial-like (NRK-52E) cells following treatment with a high concentration of calcium, while the knockout or overexpression of MMP-9 could, respectively, significantly inhibit or upregulate the expression of osteoblastic-related proteins and calcium crystal deposition. In addition, apoptosis and calcium crystal deposition were significantly reduced in Sprague–Dawley rats with 1,25(OH)2D3-induced hypercalciuria following MMP-9 inhibitor I treatment. Furthermore, inhibiting reactive oxygen species (ROS) production or the nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) pathway significantly reduced calcium-induced MMP-9 expression and calcium crystal deposition. In summary, our results suggested that a high calcium concentration promotes epithelial–osteoblastic transformation and calcium crystal deposition in renal tubule cells by regulating the ROS/NF-κB/MMP-9 axis and identified a novel role for MMP-9 in regulating calcium-induced calcium crystal deposition in renal tubules.

MicroRNA-1253 Suppresses Cell Proliferation Migration and Invasion of Osteosarcoma by Targeting MMP9.

Purpose:MicroRNAs play an important role in osteosarcoma (OS) development and progress. Although miR-1253 was considered as a tumor-inhibitor in some cancers, it’s function in the OS is not clear.Methods:In our study, we examined the expression of miR-1253 in OS cells and osteoblast cells using quantitative real-time PCR. The proliferation of OS cells was measured by BrdU assay, and we performed transwell to detect migration and invasion of OS cells. Meanwhile, EMT proteins were tested by western blot. We used Bioinformatics to predict the target genes of miR-1253 and found out Matrix metalloproteinases9 (MMP9) was one of that. The direct combination between miR-1253 and MMP9 was verified by double luciferase reporting experiment. Quantitative real-time PCR and western blot were used to detect the expression of MMP9.Results:We found that the expression level of miR-1253 in OS cells was significantly lower than that in osteoblast cells. Overexpression of miR-1253 could significantly inhibit OS cell proliferation, migration, invasion and EMT. And then, MMP9 was predicted as a downstream target of miR-1253 by Bioinformatics analysis. Further experiments showed that miR-1253 could reduce the protein level of MMP9 by directly binding to the 3’-UTR of MMP9. Afterward, we performed a rescue experiment, in which both MMP9 and miR-1253 were overexpressed. Compared with the groups overexpressed miR-1253 alone, cell proliferation, migration and invasion in co-overexpression groups were improved.Conclusions:In summary, these results suggested that miR-1253 down-regulated in OS cells, and could suppress the proliferation, migration and invasion of OS cells. Its molecular regulatory mechanism was that inhibits the expression of the downstream target gene MMP9 by directly binding, thus affect OS cell functions. Therefore, miR-1253 has the potential to become a biomarker and therapeutic target for OS therapy.

Expression of matrix metalloproteinase-9 in gingival tissue biopsy in patients with slowly/ moderately and rapidly progressing periodontitis: An observational study.

Background:Matrix metalloproteinases (MMPs) are a group of host-derived zinc-dependent enzymes which mediates the destruction of the extracellular matrix. In periodontitis, there is excess production of MMPs associated with periodontal tissue destruction. The aim of this study was to estimate the level MMP-9 in both active and latent form in gingival tissue (GT) samples collected from periodontitis patients with different rates of progression and compare it with healthy individuals.Materials and Methods:Sixty patients were selected and divided into three groups, 20 each: Group A (slowly/moderately progressing periodontitis), Group B (rapidly progressing periodontitis), and Group C (clinical periodontal health). Plaque index, gingival index, periodontal probing depth (PPD), and clinical attachment level were recorded. GT samples were collected from all 60 patients and MMP-9 expressions were measured using gelatin zymography and western blotting.Results:Levels of active MMP-9 (aMMP-9) and latent MMP-9 (lMMP-9) were significantly high in both Group A (GA) (aMMP-9: 2.05 arbitrary unit [AU]/lMMP-9: 2.54 AU) and Group B (GB) (aMMP-9: 1.32 AU/lMMP-9: 1.74 AU) when compared to that of Group C (GC) (aMMP-9: 0.93/lMMP-9: 1.08 AU). In GA, levels of aMMP-9 showed a significant correlation with PPD values. No other correlations were found.Conclusion:The levels of aMMP-9 and lMMP-9 were increased in both the types of periodontitis when compared with periodontally healthy individuals. A significant correlation was found between PPD and activities of aMMP-9 in slowly/moderately progressing periodontitis patients. However, further studies are required to confirm these findings.

Effects of Plumbagin on Proliferation, Invasion and Migration of Glioma Cell Line U-118

To observe the inhibitory effect of plumbagin on the invasion and migration of glioma cells U-118 and to explore the possible mechanism of its action. U-118 cells were cultured in medium with different concentrations of plumbagin and the cells were counted and morphological changes were observed under the microscope after 24 h of incubation and the cell proliferation was detected by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide, the effect of plumbagin on cell migration was detected by cell scratching assay and the effect of cell invasion was detected by Transwell method. Western blotting was used to detect the expression of matrix metalloproteinase-2, matrix metalloproteinase-9 and p21 protein. The results showed that the morphology of U-118 cells could be changed by plumbagin and it was dependent on the concentration; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide results showed that plumbagin significantly inhibited the growth of U-118 cells and its inhibitory ability was positively correlated with the concentration; scratch assay results showed that the planar motility of cells decreased significantly with the increase of plumbagin concentration; Transwell Western blotting results showed that plumbagin administration could inhibit the expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 proteins in U-118 cells, but significantly increased the expression of P21 and there was a positive correlation with the concentration. Plumbagin can inhibit the proliferation of U-118 cells and suppress cell invasion and the mechanism may be related to the inhibition of matrix metalloproteinase-2, matrix metalloproteinase-9 and p21 expression.

An aqueous pomegranate peel extract (Punica granatum) protect against Elastase-induced pulmonary emphysema in Sprague Dawley rats model

We investigated the effect of Punica granatum peel aqueous extract (PGE), on pulmonary inflammation and alveolar degradation induced by intratracheal administration of Elastase in Sprague Dawley rats. Lung inflammation was induced in rats by intratracheal instillation of Elastase. On day 1 and 2, animals received an intraperitoneal injection of PGE (200 mg/mL), three hours later, they were intratracheally instilled with 25U/kg pancreatic porcine Elastase. Animals were sacrificed 7 days later. Bronchoalveolar lavage (BAL) were collected and cellularity, histology and mRNA expression of Monocyte chemotactic protein 1(MCP-1), Tumor Necrosis Factor-Alpha (TNF-α), Interleukin 6 (IL-6), and Matrix Metalloproteinase-2 (MMP-2) were studied. In addition, activity of TNF- α, IL-6 and MCP-1 on BAL were also analyzed by ELISA Kit. Elastase administration increased: BAL cellularity, neutrophils recruitment and BAL MCP1, IL-6 expressions. It also increased lung TNF-α, MCP-1, MMP-2 expressions, platelets recruitment, histological parameters at 7th day of elastase treatment. Intraperitoneal injection of 200 mg/kg of PGE reduced, significantly, BAL cellularity, and neutrophils recruitment. However, in animal treated with PGE, MCP-1, MMP-2 and IL-6 on day 7, were similar to the Sham group. Treatment with PGE (200 mg/ kg) also significantly reduced lung TNF-α, and MCP-1 expression. This study reveals that PGE Punica granatum protects against elastase lung inflammation and alveolar degradation induced in rats.