Matrix Metalloproteinase-9 mRNA Expression Levels Research Articles

Injection of Matrix Metalloproteinase-9 Leads to Ventricular Remodeling.

Objective Previous studies have found that some ventricular remodeling is accompanied by increased matrix metalloproteinase-9 (MMP-9) in vivo, and MMP-9 inhibitors can reduce ventricular remodeling. However, there is still no direct evidence that MMP-9 causes ventricular remodeling. In this study, MMP-9 was injected into rats to observe whether MMP-9 caused ventricular remodeling, thereby providing direct evidence of MMP-9-induced ventricular remodeling. Methods Forty-eight eight-week-old male Wistar rats were randomly divided, by weight, into control, low-, medium-, and high-dose MMP-9 groups and were administered normal saline or recombinant rat MMP-9 0.7, 1.4, or 2.1 ng/g, respectively, via intraperitoneal injection, twice per week. On the 28th day, six rats were randomly selected from each group (Stage I). The remaining rats continued receiving injections until the 56th day (Stage II). Echocardiography was performed to observe cardiac structure and function, and the left ventricular mass index (LVWI) was calculated. Myocardial pathological changes and the collagen volume fraction (CVF) were observed by HE and VG staining in myocardial tissue. MMP-9 levels in serum were tested using ELISA. Myocardial MMP-9 levels were measured using Western blots, and the myocardial expression levels of MMP-9 mRNA were assessed using RT-PCR. Results During Stage I, serum MMP-9 and myocardial MMP-9 mRNA levels are increased; hypertrophic cardiomyocytes, disorderly arrangement of fibers, and endochylema dissolution are observed in the medium- and high-dose groups. The left ventricular weight index (LVWI) and myocardial MMP-9 increased, and the collagen volume fraction (CVF) reduced in the high-dose group. In Stage II, the left ventricular end-diastolic volume (LVEDV) and diameter (LVIDd) are higher, and CVF decreased in the medium- and high-dose groups. Myocardial pathological lesions intensified. Serum MMP-9 in the model groups and myocardial MMP-9 in the medium- and high-dose groups are increased. Conclusions Injection of MMP-9 can lead to ventricular remodeling.

Can serum matrix metalloproteinase-9 and SMAD-2 levels predict lamina propria invasion in bladder urothelial carcinoma?

Matrix metalloproteinase-9 (MMP-9) has a role in the destruction of lamina propria (LP) of the bladder wall and SMAD-2 promotes cell-to-cell adhesion. This study aimed to investigate the association between LP invasion and serum protein and mRNA expression levels of MMP-9 and SMAD-2 in bladder cancer (BC) patients. Serum samples were taken from 57 patients with suspicious BC before TUR-BT (Group 1) and 20 patients with benign diseases as control (Group 2). The mRNA expression and serum protein levels of MMP-9 and SMAD-2 were analyzed using Real-Time PCR and ELISA methods, respectively. The comparison of protein and mRNA expression levels of MMP-9 and SMAD-2 were done statistically between Group 1 and 2, as well as for different T stages of BC. The protein levels of MMP-9 (2448 vs 637.5pg/mL, P=.0001) and SMAD-2 (6.85 vs 1.61 P=.0001) were significantly higher in Group 1 compared to Group 2. The mRNA expression levels of MMP-9 (P=.89) and SMAD-2 (P=.99) did not significantly differ between the groups. The protein levels of MMP-9 in T1 patients were significantly higher from both of pTa patients (P=.018) and pT2 (P=.02). The protein levels of SMAD-2 were not statistically different between T stages. Similarly, the mRNA expression levels of MMP-9 and SMAD-2 were not different between T stages. The protein levels of MMP-9 and SMAD-2 were increased in BC patients while mRNA expressions were not different. Furthermore, the increased protein level of MMP-9 in T1 patients was more pronounced which may be related to LP invasion of the tumor.

Fullerenol Nanoparticles Decrease Blood-Brain Barrier Interruption and Brain Edema during Cerebral Ischemia-Reperfusion Injury Probably by Reduction of Interleukin-6 and Matrix Metalloproteinase-9 Transcription

BackgroundThe present study aimed to examine the protective role of fullerenol nanoparticles against blood-brain barrier (BBB) interruption and brain edema during cerebral ischemia-reperfusion injury probably by reduction of interleukin-6 (IL-6) and matrix metalloproteinase-9 (MMP-9) transcription. MethodsThe male Wistar rats (weighting 280-320 g) were randomly assigned into four groups as follows: sham, control ischemic, pretreated ischemic, and posttreated ischemic groups. Cerebral ischemia-reperfusion (IR) injury was performed by occlusion of middle cerebral artery (MCA) for 90 minutes followed by twenty-four hours reperfusion. Rats were administered fullerenol 5mg/kg, intraperitoneally, 30 minutes before induction of IR in pretreated ischemic group and immediately after termination of MCA occlusion in posttreated ischemic group. After twenty-four hours reperfusion, the method of Evans blue dye extravasation (EBE) and RT-PCR were used for determination of BBB permeability and mRNA expression levels of MMP-9 and IL-6, respectively. Neuronal deficit score (NDS) and edema of the ischemic hemispheres were also evaluated. ResultsMCA occlusion increased NDS in control ischemic rats (3.16 ± 0.16) with concomitant increase in EBE (15.30 ± 3.98µg/g) and edema (3.53 ± 0.50%). Fullerenol in both pretreated and posttreated ischemic groups reduced NDS (36% and 68%, respectively), EBE (89% and 91%, respectively) and edema (53% and 81%, respectively). Although MCA occlusion increased the mRNA expression levels of MMP-9 and IL-6 in ischemic hemispheres, fullerenol in both treatment groups noticeably decreased the mRNA expression levels of these genes. ConclusionIn conclusion, fullerenol nanoparticles can protect BBB integrity and attenuate brain edema after cerebral ischemia-reperfusion injury possibly by reduction of IL-6 and MMP-9 transcription.

Inhibition of MMP-9 using a pyrrole-imidazole polyamide reduces cell invasion in renal cell carcinoma

We investigated the clinical significance of the expression levels of matrix metalloproteinase 9 (MMP-9) in renal cell carcinoma (RCC). In addition, we validated the efficacy of pyrrole imidazole polyamide (PIP) targeting MMP-9 on inhibiting proliferation and invasion of RCC. We evaluated the expression levels of MMP-9 in 249RCC specimens by immunostaining and analyzed the association between MMP-9 expression levels and cancer-specific survival. Furthermore, in a human RCC cell line, Caki-2, we tested the effect of a couple of PIPs targeting MMP-9 one recognizing an NF-κB binding site (MMP-9-NF-κB PIP) and another for the AP-1 binding site (MMP-9-AP-1 PIP) in the MMP-9 promoter. The expression levels of MMP-9, proliferative activity and invasive capability were tested by quantitative PCR, WST8 assay and Matrigel invasion assay, respectively. By immunostaining of the clinical specimens, strong MMP-9 staining was proven to be a significant predictor of poor prognosis for cancer-specific survival (P<0.01). In Caki-2 cells, MMP-9-NF-κB PIP significantly reduced the expression levels of MMP-9 mRNA and inhibited cell invasion, but did not affect the cell proliferation activity. On the other hand, no effect was found in MMP-9-AP-1 PIP on MMP-9 mRNA expression, cell proliferation and invasion. We confirmed the inhibitory effects of MMP-9-NF-κB PIP on the expression of MMP-9 and subsequent invasion of Caki-2 cells. Since it was clearly shown that high MMP-9 expression levels were associated with poor prognosis of RCC, MMP-9 is a potential candidate target for RCC treatment. Transcription therapy using a minor groove binder, such as NF-κB PIP, may be a potential therapeutic agent for RCC, although further investigation is required.

Derlin-1 Is Overexpressed in Non-Small Cell Lung Cancer and Promotes Cancer Cell Invasion via EGFR-ERK–Mediated Up-Regulation of MMP-2 and MMP-9

Previous studies indicated a role of Derlin-1 in human cancers; however, its expression pattern in non-small cell lung cancer (NSCLC) and the molecular mechanism of Derlin-1 on cancer progression have not been characterized. In the present study, Derlin-1 expression was examined in lung cancer cell lines and human tissues. Derlin-1 overexpression correlated with pTNM stage, lymph node metastasis, and poor overall survival. siRNA knockdown of Derlin-1 impaired anchorage-dependent and anchorage-independent cell growth and invasion in A549 and H1299 cell lines, and its overexpression promoted proliferation and invasion in HBE and LTE cell lines. Derlin-1 depletion decreased matrix metalloproteinase (MMP)-2/9 at both protein and mRNA levels, with decreased MAP kinase/extracellular signal-regulated kinase (ERK)/ERK phosphorylation. Derlin-1 overexpression up-regulated MMP-2/9 expression and ERK phosphorylation, which could be reversed by MAP kinase/ERK kinase inhibitor, PD98059. The effect of Derlin-1 on MMP-2/9 up-regulation was abolished in ERK1/2 siRNA-treated cells. Further analysis showed that Derlin-1 overexpression induced EGFR phosphorylation. EGFR inhibitor blocked Derlin-1-mediated up-regulation of EGFR and ERK phosphorylation. MMP-2/9 and p-ERK up-regulation by Derlin-1 was partly blocked in EGFR-depleted cells with siRNA treatment. Immunoprecipitation confirmed the association between Derlin-1 and EGFR. In summary, our results showed that Derlin-1 is overexpressed in NSCLC and promotes invasion by EGFR-ERK-mediated up-regulation of MMP-2 and MMP-9. Derlin-1 may serve as a therapeutic target for NSCLC.

Levels of Metalloproteinase (MMP-3, MMP-9), NF-kB Ligand (RANKL), and Nitric Oxide (NO) in Peripheral Blood of Osteoarthritis (OA) Patients

The aim of this study was to judge whether there is a correlation between some biochemical features of knee osteoarthritic blood and clinical characteristics and to evaluate the potential relationship between osteoarthitis (OA) severity and putative biomarkers for the disease. 105 patients suffering from knee OA were analyzed clinically (Lequesne's index) and radiographically (Kellgren and Lawrence, K&L). Plasma and peripheral blood mononuclear cells (PBMC) were harvested separately. Specimens were analyzed for concentrations of nitric oxide (NO), matrix metalloproteinase-3 (MMP-3), and matrix metalloproteinase-9 (MMP-9). Transcript levels of the receptor activator of NF-kB ligand (RANKL) mRNA, MMP-3mRNA, and MMP-9mRNA were measured using real-time quantitative RT-PCR. Data certified significantly increasing concentrations of plasma MMP-3, MMP-9, and NO as well as transcript levels of RANKL mRNA and MMP-9 mRNA in early OA (at grade I). There was a positive correlation of MMP-3 and MMP-9 content in plasma and MMP-9 mRNA expression levels in PBMC with the severity of clinical symptoms (total Lequesne's scores) in early OA. NO content in plasma correlated with total Lequesne's scores, pain scores in response to pressure, and swelling scores of early OA patients (atgGrade I). Analogously, there were positive correlations of RANKL mRNA expression with total Lequesne's scores, pain scores in response to pressure, and swelling scores in OA patients at Grade I. [corrected] Some biochemical factors, including content of NO, MMP-3, MMP-9, and transcript levels of some genes, including MMP-9 mRNA and RANKL mRNA, may be specific and sensitive enough to diagnose OA diseases at an early stage in the pathological process of OA when radiological features do not reflect degradation of articular cartilage. Therefore, proper regulation of these factors may be a promising and realistic new target for the treatment of degenerative osteoarticular diseases.

Dimethyl Sulfoxide Attenuates TNF-α-Induced Production of MMP-9 in Human Keratinocytes

Dimethyl sulfoxide (DMSO), an aprotic solvent, is found to be useful as a topical agent with antioxidant effects in treatment of chronic wounds. However, the effects of DMSO on matrix metalloproteinase-9 (MMP-9) production in the presence of an inflammatory environment as in the case of disordered wound healing has not been previously investigated. The aim of this study was to investigate whether TNF-α-induced MMP-9 levels and MMP-9 mRNA expression from human keratinocytes (HaCaT) might be attenuated by DMSO. Human keratinocytes were treated with DMSO (0.1–1%) for 24 h and then exposed to tumor necrosis factor (TNF)-α (10 ng/ml) for an additional 24 h. Expression and production of MMP-9 from HaCaT cells were determined by reverse transcription polymerase chain reaction (RT-PCR) and gelatin zymography, respectively. Results showed that DMSO inhibited production of both MMP-9 levels and MMP-9 mRNA expression in TNF-α-stimulated cells in a concentration-dependent manner. Inhibition of MMP-9 levels was statistically significant at DMSO concentrations of 0.75% and higher. Similarly, the increase of MMP-9 mRNA expression levels in TNF-α-stimulated cells was markedly reduced by DMSO. Data suggest that DMSO may attenuate the deleterious effects of MMP-9 through downregulation at the transcription level. Therefore, DMSO may provide a good strategy to prevent TNF-α-induced proteolytic activity in cutaneous inflammatory reactions.

Suppression of phorbol-12-myristate-13-acetate-induced tumor cell invasion by apigenin via the inhibition of p38 mitogen-activated protein kinase-dependent matrix metalloproteinase-9 expression

Apigenin has special interest for the development of chemopreventive agents against cancer because it is a widely distributed plant flavonoid that has antitumor properties. In this study, we investigated the apigenin effects on the protease-mediated invasiveness in human metastatic cancer cell lines Caski, SK-Hep1, and MDA-231. We found that apigenin markedly inhibits the phorbol-12-myristate-13-acetate (PMA)-induced increase in MMP-9 expression and activity in several cancer cell lines. These effects of apigenin are dose-dependent and correlate with the suppression of MMP-9 mRNA expression levels. PMA caused about a 5-fold induction in MMP-9 promoter activity, which was also suppressed by apigenin treatment in Caski cells. We found that apigenin could inhibit PMA-induced phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK), which was involved in the down-regulation of the expression of matrix metalloproteinase-9 (MMP-9) at mRNA levels. Furthermore, the treatment of inhibitors specific for p38 MAPK (SB203580) to Caski cells caused the reduction of MMP-9 expression. Restoration of p38 expression partly increased PMA-induced MMP-9 secretion blocked by apigenin treatment in Caski cells. These results showed apigenin might inhibit the invasion and migration abilities of Caski cells by reducing the MMP-9 expression through suppressing the p38 MAPK signaling pathway. These findings indicate that apigenin might be a useful strategy for controlling metastasis and the invasiveness of tumors.

Matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) mRNA expression in patients with metastatic gastric cancer receiving first-line chemotherapy

4547 Background: In preclinical tumor models, VEGF and MMP-9 induced each other, resulting in the enhancement of tumor angiogenesis and the formation of metastases. This study evaluated MMP-9 and VEGF in patients (pts) with metastatic gastric cancer. Methods: Patient tumor samples from a phase III trial of the AIO were prospectively collected. VEGF and MMP-9 mRNA were isolated from paraffin embedded tissues using a new patented method based on nanotechnology. The expression was then assessed by qRT-PCR and by immunohistochemistry (IHC). Results: A total of 137 (training cohort 96; validation cohort 41) out of 220 pts enrolled were analyzed. In the training cohort, increased MMP-9 levels were strongly associated with increased VEGF expression, resistance to chemotherapy and shorter progression-free and overall survival. At the cut-off with the highest predictive value, the rate of disease progression as best response associated with elevated MMP9 expression was 50% (vs. 22.4%; Fisher's test p=.027) and median overall survival (OS) was 3.1 months (vs. 8.7 months, log rank p=.0000165). This prognostic value for survival was maintained when data were analyzed in the multivariate setting (relative hazard ratio for death 3.67, p=.00045) or were categorized according to the quartile (p=.004) or median (p=.024) distributions for MMP-9. VEGF showed a prognostic effect in pts with low MMP-9 expression, only. In the validation cohort, the prognostic value of increased MMP-9 expression could be confirmed at the best cut off derived from the training cohort, with the hazard ratio for death being 3.45 (p=.02). Interestingly, in the IHC, stromal but not tumoral MMP-9 protein expression predicted survival (log rank p=.033). Conclusions: Tumoral MMP-9 mRNA expression levels are the first validated molecular prognostic factor in metastatic gastric cancer. These results have implications for novel anti-protease/ anti-VEGF treatment strategies. [Table: see text]

Association of elevated matrix metalloproteinase-9 (MMP-9) mRNA expression levels with resistance to chemotherapy and survival in patients with metastatic gastric cancer receiving first-line chemotherapy: Results from the FLO versus FLP gastric cancer phase III trial of the AIO

4544 Background: MMP-9 has been implicated in invasion and metastasis of cancer, but little is known about its prognostic role in patients (pts) with metastatic gastric cancer undergoing chemotherapy. Methods: Tumor samples from pts who received first-line 5-fluorouracil, leucovorin and platinum for metastatic gastric cancer within a phase III trial of the AIO were prospectively collected and analyzed. The expression of MMP-9 mRNA was assessed by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Moreover, EGFR, pAKT, pMAPK, HER2/neu, Ki-67, CD8+ cell and macrophage accumulation and VEGF-A were evaluated (by qRT-PCR and/or immunohistochemistry). Results: Overall, 288 samples from 168 out of 220 pts enrolled were analyzed for at least one of the marker. 158 samples of 109 pts have been assessed for MMP9 mRNA so far; 96 pts were eligible for the analysis. In this cohort, increased MMP-9 levels were strongly associated with resistance to chemotherapy and shorter progression-free and overall survival. At the cut-off with the highest predictive value, the rate of disease progression as best response associated with elevated MMP9 expression was 50% (versus 22.4%; fisher’s test p=0.0276) and the relative hazard ratio for death was 2.653 (95% CI, 2.060 - 9.268); log rank p<.0001). The prognostic value for survival was maintained when data were categorized according to the median (p=.024) or quartile (p=0.004) distributions for MMP-9 expression. Interestingly, elevated MMP-9 levels were associated with more lymph node and peritoneal and less liver involvement and correlated with increased intratumoral VEGF-A levels. Other correlative analyses will be presented at the meeting. Conclusions: The association of MMP-9 expression with poor response to chemotherapy, and shorter survival suggests that MMP-9 is linked not only to early invasion and metastases formation, but also to an aggressive clinical course in metastatic disease. Thus, MMP-9 mRNA expression levels may represent a valuable marker for novel anti-protease treatment strategies. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration Siemens