Matrix Metalloproteinase 14 Overexpression Research Articles

CLDN1 silencing suppresses the proliferation and migration of airway smooth muscle cells by modulating MMP14

Airway remodeling is an important pathologic factor in the progression of asthma. Abnormal proliferation and migration of airway smooth muscle cells (ASMCs) are important pathologic mechanisms in severe asthma. In the current study, claudin-1 (CLDN1) was identified as an asthma-related gene and was upregulated in ASMCs stimulated with platelet-derived growth factor BB (PDGF-BB). Cell counting kit-8 and EdU assays were used to evaluate cell proliferation, and transwell assay was carried out to analyze cell migration and invasion. The levels of inflammatory factors were detected using enzyme-linked immunosorbent assay. The results showed that CLDN1 knockdown inhibited the proliferation, migration, invasion, and inflammation of ASMCs treated with PDGF-BB, whereas overexpression of CLDN1 exhibited the opposite effects. Protein-protein interaction assay and co-immunoprecipitation revealed that CLDN1 directly interacted with matrix metalloproteinase 14 (MMP14). CLDN1 positively regulated MMP14 expression in asthma, and MMP14 overexpression reversed cell proliferation, migration, invasion, and inflammation induced by silenced CLDN1. Taken together, CLDN1 promotes PDGF-BB-induced cell proliferation, migration, invasion, and inflammatory responses of ASMCs by upregulating MMP14 expression, suggesting a potential role for CLDN1 in airway remodeling in asthma.

LongShengZhi alleviated cardiac remodeling via upregulation microRNA-150-5p with matrix metalloproteinase 14 as the target

Ethnopharmacological relevanceLongShengZhi capsule (LSZ), a traditional Chinese medicine, is used for treatment of patients with vascular diseases. LSZ reduced doxorubicin-induced heart failure by reducing production of reactive oxygen species and inhibiting inflammation and apoptosis. Aim of the studyThis study was to explore whether LSZ could alleviate cardiac remodeling via upregulation of microRNA (miR)-150-5p and the downstream target. Cardiac remodeling was induced by Ang II in vivo and in vitro. ResultsLSZ attenuated Ang II-induced cardiac hypertrophy and fibrosis in rats, and in primary cardiomyocytes (CMs) and primary cardiac fibroblasts (CFs). MiR-150-5p was downregulated in Ang II-induced rat heart, CMs and CFs, and these decreases were reserved by LSZ. In vivo overexpression of miR-150-5p by transfection of miR-150-5p agomiR protected Ang II-induced cardiac hypertrophy and fibrosis in rats. Meanwhile, its overexpression also reversed Ang II-induced upregulation of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) in rat hearts and primary CMs, as well as upregulation of collagen I, collagen III and transforming growth factor-β (TGF-β) in rat hearts and primary CFs. Matrix metalloproteinase 14 (MMP14) was validated as the target gene of miR-150-5p, which was overexpressed in Ang II-induced rat heart, rat primary CMs and primary CFs. Notably, overexpression of MMP14 induced cardiac remodeling, and reversed the protective role of miR-150-5p in downregulating Ang II-induced upregulation of hypertrophy and fibrosis markers in vitro. ConclusionCollectively, LSZ protects Ang II-induced cardiac dysfunction and remodeling via upregulation of miR-150-5p to target MMP14. Administration of LSZ, upregulation of miR-150-5p or targeting of MMP14 may be strategies for cardiac remodeling therapy.

Overexpression of MMP14 predicts the poor prognosis in gastric cancer: Meta-analysis and database validation.

Background:Plenty of studies have showed matrix metalloproteinase 14 (MMP14) expression might be associated with the prognosis of gastric cancer (GC). However, no definite conclusion has been obtained for the contradictory results.Methods:We searched PubMed, Web of science, Embase, and Cochrane library for eligible studies. The association between MMP14 expression and prognostic outcomes of GC was evaluated. Hazard ratio (HR) and 95% confidence interval (CI) were integrated to show the effect of MMP14 expression on the overall survival (OS) or recurrence-free survival (RFS). Data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) was used to validate the association of MMP14 expression with OS or RFS in GC. A brief bioinformatics analysis was also performed to determine the prognostic role of MMP14 expression in GC.Results:High MMP14 expression was associated with shorter OS compared to low MMP14 expression in GC (HR = 1.95, P < .01). Patients with high MMP14 expression tended to have worse differentiation (P = .03), deeper tumor invasion (P < .01), earlier lymph node metastasis (P < .01), earlier distant metastasis (P < .01) and more advanced clinical stage (P < .01) compared to those with low MMP14 expression. The data from TCGA and GEO showed MMP14 was overexpressed in tumor tissues compared to normal tissues (P < .05), and high MMP14 expression was significantly related to shorter OS (HR = 1.70, 95% CI = 1.32–2.20, P < .01) and RFS (HR = 1.45, 95% CI = 1.15–1.83, P < .01) compared to low MMP14 expression in GC. Expression of MMP14 was linked to functional networks involving the biological process, metabolic process, response to stimulus, cell communication and so on. Functional network analysis suggested that MMP14 regulated the protein digestion and absorption, extracellular matrix receptor interaction, focal adhesion, ribosome, spliceosome, and so on.Conclusion:High MMP14 expression was associated with worse prognosis of GC compared to low MMP14 expression. MMP14 expression could serve as a prognostic factor and potential therapeutic target of GC.

Effects of miR-7 on Hcy-induced rat cerebral arterial vascular smooth muscle cell proliferation, migration and inflammatory factor expression by targeting MMP-14 to regulate TLR4/NF-κB signaling pathway

The current research aimed to investigate the effect of miR-7 targeting matrix metalloproteinase 14 (MMP-14) on homocysteine (Hcy)-induced rat cerebral artery vascular smooth muscle cells (VSMCs) proliferation, migration and inflammatory factor expression and its possible mechanism. The expression of miR-7 and MMP-14 in Hcy-induced VSMCs were detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blot. Methyl Thiazolyl Tetrazolium (MTT) method, Transwell assays and enzyme-linked immunosorbent assay (ELISA) were performed to detect the effect of miR-7 and MMP-14 expression on the proliferation and migration, as well as interleukin 6 (IL-6) and tumor necrosis factor ɑ (TNF-ɑ) expression of Hcy-induced VSMCs. The interaction between miR-7 and MMP-14 was detected by dual-luciferase reporter gene assay. Western blot was applied to analyse the effects of miR-7 and MMP-14 expression on the Toll-like receptor (TLR4)/nuclear transcription factor-KB (NF-κB) signaling pathway. The results showed that after induced by Hcy, the expression of miR-7 in VSMCs was significantly reduced, the expression of MMP-14 was significantly increased, and the cell viability, the number of migrating cells, IL-6 and TNF-ɑ expression were significantly increased (P&lt;0.05). After overexpression of miR-7, the viability, migration cell numbers, IL-6 and TNF-ɑ expression of Hcy-induced VSMCs were significantly reduced (P&lt;0.05). miR-7 directly binds to MMP-14 and negatively regulates the expression of MMP-14. After overexpression of miR-7, the levels of TLR4 and p-NF-κB p65 in VSMCs were significantly reduced (P&lt;0.05); overexpression of MMP-14 could reduce the effect of miR-7 overexpression on TLR4 and p-NF-κB p65 expression in VSMCs (P&lt;0.05). Overexpression of MMP-14 and/or activation of the TLR4/NF-κB signaling pathway could reverse the effect of miR-7 overexpression on the proliferation, migration and IL-6 and TNF-ɑ expression of Hcy-induced VSMCs (P&lt;0.05). It is concluded that miR-7 can inhibit Hcy-induced rat cerebral artery VSMCs proliferation, migration, and inflammatory factor expression by targeting the regulation of MMP-14 expression and inhibiting the activation of the TLR4/NF-κB signaling pathway.

Articles of Significant Interest in This Issue

Matrix metalloproteinase 14 (MMP14) is a pericellular collagenase expressed at high levels in adipose tissue. Li et al. (e00564-19) used an adipose tissue-specific and doxycycline-inducible MMP14 overexpression model to study the effects of this factor in obese mice. They found that overexpressing MMP14 in established obese fat leads to unhealthy metabolic effects via the production of endotrophin, a known fibrotic factor that triggers local inflammation. They further revealed that hypoxia-induced hypoxia-inducible factor 1α binds to the MMP14 promoter to upregulate MMP14 expression in obese fat. These findings suggest that MMP14 may be a key player in adipose tissue remodeling during the development of obesity.

Critical Role of Matrix Metalloproteinase 14 in Adipose Tissue Remodeling during Obesity.

Fibrosis is recognized as the major pathological change in adipose tissue during the development of obesity. However, the detailed mechanisms governing the interactions between the fibrotic components and their modifiers remain largely unclear. Here, we reported that matrix metalloproteinase 14 (MMP14), a key pericellular collagenase, is dramatically upregulated in obese adipose tissue. We generated a doxycycline-inducible adipose tissue-specific MMP14 overexpression model to study its regulatory function. We found that overexpression of MMP14 in the established obese adipose tissue leads to enlarged adipocytes and increased body weights in transgenic mice. Furthermore, the mice exhibited decreased energy expenditure, impaired lipid metabolism, and insulin resistance. Mechanistically, we found that MMP14 digests collagen 6α3 to produce endotrophin, a potent costimulator of fibrosis and inflammation. Unexpectedly, when overexpressing MMP14 in the early-stage obese adipose tissue, the transgenic mice showed a healthier metabolic profile, including ameliorated fibrosis and inflammation, as well as improved lipid and glucose metabolism. This unique metabolic phenotype is likely due to digestion/modification of the dense adipose tissue extracellular matrix by MMP14, thereby releasing the mechanical stress to allow for its healthy expansion. Understanding these dichotomous impacts of MMP14 provides novel insights into strategies to treat obesity-related metabolic disorders.

Prognostic Significance of Matrix Metalloproteinase 14 in Patients with Cancer: a Systematic Review and Meta-Analysis.

There is increasing evidence that matrix metalloproteinase 14 (MMP-14) is involved in tumor progression and prognosis. MMP-14 exhibits different expression in patients with various cancers, suggesting that it may be considered as a potential prognostic biomarker for cancer. Therefore, this meta-analysis was performed to elucidate the prognostic value and association of MMP-14 over-expression in several types of cancers. Eligible studies based on eligibility criteria from various online databases were searched. The pooled hazard ratios (HRs) with 95% confidence intervals (CIs) for overall survival (OS) were analyzed to determine the prognostic value of MMP-14 using STATA software 12.0. We identified sixteen applicable studies in this meta-analysis comprising 2,766 samples. Over-expression MMP-14 was significantly correlated with a poor overall survival (OS) outcome in multiple cancers (HR: 2.22; 95% CI: 1.72 - 2.87). Moreover, high levels of MMP-14 were markedly associated with tumor progression and metastasis (HR: 1.83; 95% CI: 1.36 - 2.46). MMP-14 expression was also associated with histological differentiation (OR: 0.37; 95% CI: 0.18 - 0.77). MMP-14 over-expression suggested aggressive biological behaviors and implied that MMP-14 may be a useful prognostic biomarker in human cancers.

Downregulation of matrix metalloproteinase 14 by the antitumor miRNA, miR-150-5p, inhibits the aggressiveness of lung squamous cell carcinoma cells.

In the present study, in order to elucidate the aggressive nature of lung squamous cell carcinoma (LUSQ), we investigated the oncogenic RNA networks regulated by antitumor microRNAs (miRNAs or miRs) in LUSQ cells. The analysis of our original miRNA expression signatures of human cancers revealed that microRNA‑150‑5p (miR‑150‑5p) was downregulated in various types of cancer, indicating that miR‑150‑5p acts as an antitumor miRNA by targeting several oncogenic genes. Thus, the aims of this study were to investigate the antitumor roles of miR‑150‑5p in LUSQ cells and to identify oncogenes regulated by miR‑150‑5p that are involved in the aggressive behavior of LUSQ. The downregulation of miR‑150‑5p was validated in clinical samples of LUSQ and cell lines (SK-MES‑1 and EBC‑1). The ectopic overexpression of miR‑150‑5p significantly suppressed cancer cell aggressiveness. Comprehensive gene expression analyses revealed that miR‑150‑5p regulated 9genes in the LUSQ cells. Among these, matrix metalloproteinase 14 (MMP14) was found to be a direct target of miR‑150‑5p, as shown by luciferase reporter assay. The knockdown of MMP14 using siRNA against MMP14 (si-MMP14) significantly inhibited cancer cell migration and invasion. The overexpression of MMP14 was detected in clinical specimens of LUSQ by immunohistochemistry. On the whole, these findings suggest that the downregulation of miR‑150‑5p and the overexpression of MMP14 may be deeply involved in the pathogenesis of LUSQ.

Matrix Metalloproteinase 14 Overexpression Is Correlated with the Progression and Poor Prognosis of Nasopharyngeal Carcinoma

Matrix metalloproteinase 14 (MMP14) has been identified to play a significant role in several types of cancers, but little is known about the significance of MMP14 in nasopharyngeal carcinoma (NPC) patients. The aim of this study was to explore the association of MMP14 expression with clinicopathologic features and prognosis in NPC. MMP14 mRNA and protein expressions were examined in NPC and nasopharyngeal tissues through real-time PCR and immunohistochemistry. Meanwhile, the relationship of MMP14 expression levels with clinical features and prognosis of NPC patients was analyzed. MMP14 mRNA expression was markedly higher in NPC tissues than in nasopharyngeal epithelium tissues (p = 0.002). Using immunohistochemistry, staining for MMP14 protein was found in the normal nasopharyngeal epithelial cells and malignant epithelial cells, but increased expression of MMP14 was observed in NPC samples compared with normal nasopharyngeal epithelium samples (p = 0.027). In addition, high levels of MMP14 protein were positively correlated with the status of clinical stage (p = 0.009), N classification (p = 0.006), and distant metastasis (p = 0.005) of NPC patients. Patients with higher MMP14 expression had a significantly shorter overall survival time than did patients with low MMP14 expression. Multivariate analysis indicated that the level of MMP14 expression was an independent prognostic indicator (p < 0.001) for the survival of patients with NPC. MMP14 overexpression is a potentially unfavorable prognostic factor for NPC patients.

Increased expression of MMP14 correlates with the poor prognosis of Chinese patients with gastric cancer

The role of matrix metalloproteinase 14 (MMP14) has been identified to involve tumor progression and prognosis. The purpose of this study is to investigate the role of MMP14 in tumor progression and prognosis of gastric cancer. This study indicated that MMP14 mRNA and protein were overexpressed in gastric cancer tissue (P<0.001 and P=0.037, respectively) and significantly associated with clinical stage (P=0.005), lymph node metastasis (P=0.003), and distant metastasis (P=0.017). Moreover, we found that the overexpression of MMP14 was a significant predictor of poor prognosis for gastric cancer patients (P<0.001). Furthermore, we performed a meta-analysis which included 594 cases from 3 studies and showed that MMP14 overexpression was a significantly poor prognostic factor in Chinese patients with gastric cancer and HR (95% CI) was 2.17 (1.64–2.86). In conclusion, MMP14 plays an important role on gastric cancer progression and prognosis and acts as a convictive biomarker for prognostic prediction for Chinese patients with gastric cancer.

MMP-14 overexpression correlates with poor prognosis in non-small cell lung cancer.

Matrix metalloproteinase-14 (MMP-14) has been demonstrated to play an important role in tumor progression. The aim of this study was to analyze the correlation between MMP-14 expression and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). Immunohistochemical staining for MMP-14 protein was performed in 104 patients with NSCLC. High levels of MMP-14 protein were positively correlated with the status of clinical stage (I-II vs. III-IV; P < 0.001), N classification (N0-N1 vs. N2-N3; P < 0.001), distant metastasis (no vs. yes; P = 0.014), and differentiated degree (high vs. low or undifferentiated; P = 0.001). The patients with higher MMP-14 expression of protein had shorter survival time than patients with low MMP-14 expression. Multivariate analysis indicated that the level of MMP-14 expression was an independent prognostic indicator (P < 0.001) for the survival of patients with NSCLC. In conclusion, MMP-14 is a potential unfavorable prognostic factor for patients with NSCLC.

Matrix metalloproteinase-14 expression and its prognostic value in cervical carcinoma.

The objective of the study was to evaluate the expression of matrix metalloproteinase-14 (MMP-14) in cervical carcinoma and correlate its expression with clinicopathological parameters, recurrence, and survival of the patients. The expressions of MMP-14 in normal cervical mucosa and cervical carcinoma tissue were detected with immunohistochemistry. Survival analysis by the Kaplan-Meier method was performed to assess prognostic significance. The positive expression rate of MMP-14 in cervical carcinoma tissue was 81.6 %(111/136), and there was significant difference on their positive expression rates between in cervical carcinoma tissue and in normal cervical mucosa(22.4 %)(13/58)(P < 0.05);The positive expression rates of MMP-14 in patients with poor histologic differentiation, lymph node metastasis, and recurrence group were heightened. Using Kaplan-Meier analysis, a comparison of survival curves of low versus high expressions of MMP-14 revealed a highly significant difference in human cervical carcinoma tissue (P < 0.05), which suggests that overexpression of MMP-14 is associated with a worse prognosis. The MMP-14 which promotes angiogenes is associated with lymph node metastasis, vascular invasion, and poor prognosis of cervical carcinoma. The current study shows that MMP-14 may be an independent prognostic factor for cervical carcinoma patients.

Matrix Metalloproteinase-14 Is a Negative Prognostic Marker for Patients with Gastric Cancer

Matrix metalloproteinase-14 (MMP-14) has been considered to play an important role in invasion and metastasis of human solid tumor. The present study aimed to investigate the association of MMP-14 with overall survival in human gastric cancer. Gastric cancer and adjacent normal specimens were collected from 205 patients who had not received neoadjuvant chemotherapy. MMP-14 expression was investigated by immunohistochemistry assay and staining evaluation results were analyzed statistically in relation to overall survival of patients. MMP-14 expression proved to be increased in gastric cancer compared with that in normal tissues. It was also proved that MMP-14 expression was associated with tumor invasion, metastasis, and TNM stage while no correlations were detected between MMP-14 expression and age, sex, differentiation status, or Lauren's classification. Moreover, patients with gastric cancer of MMP-14-positive expression tend to have worse overall survival compared with those with MMP-14 negative expression. The present study confirmed the over-expression of MMP-14 in human gastric cancer and its association with tumor progression. It also provided the first evidence that MMP-14 expression in gastric cancer was an independent negative prognostic factor of patients.

Metalloproteases Could Be Involved in Erythroid Differenciation in MPN

Abstract 5161 BackgroundSeveral hypotheses have been developed to explain how a unique mutation is related to three different phenotypes in Philadephia negative myeloproliferative neoplasms (MPN). Metalloproteases are a group of proteins involved in matrix remodelling, migration and differentiation processes. Although their role has never been studied in MPN, our group has found a differential expression of Matrix metalloproteinase-14 (MMP14) and CD44 between PV and ET JAK2V617F patients (E. Albizua et al, Ann Hematol. 2011Feb). AimsThe purpose of the present study is to validate MMP14 and CD44 differential gene expression profile between PV and ET using a proteomic-based approach and an in vitro model, with the aim of confirming MMP14 and CD44 as MPN biomarkers as well as novel targeted therapy. MethodsSeventy MPN patients diagnosed by WHO criteria were included in the study: 24 Polycythemia Vera (PV) 24 Essential Thrombocythemia (ET) JAK2V617F, 11 ET JAK2 wild type (JAK2WT) and 11 Primary Myelofibrosis (PMF). In addition 24 healthy donors were used as controls.Peripheral blood was analyzed by flow cytometry (FCM) using anti-CD44 and anti-CD45 antibodies in 10 PV and 10 ET. Fifty-five bone marrow biopsies (11 PV, 11 ET JAK2V617F, 11 ET JAK2WT, 11 PMF and 11 healthy controls) were processed to perform immunohistochemistry (IHC) with anti-MMP14 and anti-CD44 antibodies (R&D). Finally an in vitro MPN model was performed. Mononuclear cells from 10 MPN (5 PV and 5 ET) and 5 healthy controls were extracted and seeded in Methocult with IL-3, SCF and EPO. MMP14 was inhibited by Marimastat at 50μM, 25μM and 10μM; and CD44, with anti-CD44 antibody at 10mg/ml, 1mg/ml and 0.1mg/ml. Results were analyzed by BFU-E count, viability study by trypan blue and FCM employing anti-CD45, anti-CD41, anti-CD34, anti-CD71, anti-CD44, and Annexin antibodies. BFU-E DNA was extracted to analyze JAK2V617F allele burden by RT-PCR.Intracellular proteins, including phospho-proteins p38, P-p38, MEK, P-MEK, STAT1, P-STAT1, AKT and P-AKT were studied by cytometric bead array multiplexed bead-based immunoassay (CBAs) technique.The Mann-Whitney non-parametrical statistical hypothesis test was used to assess the statistical significance of our results. ResultsFirst of all, FCM of the granulocytes showed a slight over-expression of CD44 in PV population versus ET.Secondly, IHC also showed MMP14 over-expression in megakaryocites of PV (100% positive patients, 60% positive megakaryocites), and ET (100% positive patients 73% positive megakaryocites) compared to controls (36% positive subjects, 2% positive megakaryocites). Additionally 45% ET JAK2WT were MMP14 positive (20% positive megakaryocites).Significant differences in inhibition of BFU-E growth and cell proliferation were found comparing cultures either under anti-MMP14, Marimastat (50 μM and 25 μM) or under anti-CD44 (10 mg/ml and 1 mg/ml-) to cultures under no inhibitory treatment (P=0.05 and P=0.037 respectively). FCM of BFU-E cultures pointed to a significant decrease of CD71 (erythroid population) with any of the inhibitors.Over-expression of CD44 in PV compared to ET was observed and confirmed in BFU-E by FCM.All phospho-proteins studied halved after anti-CD44 (1 mg/ml) and Marimastat treatments (25 μM). ConclusionsOur results suggest that MMP14 and CD44 could play a role in erythroid differentiation by JAK-STAT, MAPK and AKT pathways. Differences between ET and PV may be caused by both molecules, which may contribute to phenotypic divergence. Additionally, MMP14 IHC could become a useful diagnostic tool in, at least, 45% of ET JAK2WT. Our results suggest that MMP14 and CD44 could become future therapeutic targets and could help to MPN diagnosis. Disclosures:No relevant conflicts of interest to declare.

Matrix metalloproteinase 14 overexpression reduces corneal scarring

Once a corneal scar develops, surgical management remains the only option for visual rehabilitation. Corneal transplantation is the definitive treatment for a corneal scar. In addition to the challenges posed by graft rejections and other postoperative complications, the lack of high-quality donor corneas can limit the benefits possible with keratoplasty. The purpose of our study was to evaluate a new therapeutic strategy for treating corneal scarring by targeting collagen deposition. We overexpressed a fibril collagenase (matrix metalloproteinase 14 (MMP14)) to prevent collagen deposition in the scar tissue. We demonstrated that a single and simple direct injection of recombinant adeno-associated virus-based vector expressing murine MMP14 can modulate gene expression of murine stromal keratocytes. This tool opens new possibilities with regard to treatment. In a mouse model of corneal full-thickness incision, we observed that MMP14 overexpression reduced corneal opacity and expression of the major genes involved in corneal scarring, especially type III collagen and α-smooth muscle actin. These results represent proof of concept that gene transfer of MMP14 can reduce scar formation, which could have therapeutic applications after corneal trauma.