BACKGROUND/AIMS: Matrix metalloproteinases (MMP)-2 and -9 can degrade essential components of vascular integrity. The aim of this study was to investigate the association between those MMPs and variceal bleeding (VB). METHODS: Fifteen controls, 12 patients with acute ulcer bleeding (UB) group, 37 patients with varix (V group), and 35 patients with acute VB group were enrolled. Serum was obtained to measure MMP-2 and -9 activity by zymogram protease assays. RESULTS: The activity levels of these compounds were compared with the controls' median value. The median MMP-9 activity was 1.0 in controls, 1.05 in the UB group, 0.43 in the V group, and 0.96 in the VB group. The level of MMP-9 activity was higher in the VB group than in the V group (p<0.001). In the VB group, there was a signifi cant decrease in MMP-9 activity over time after bleeding (p<0.001). The median MMP-2 activity level was 1.0 in controls, 1.01 in the UB group, 1.50 in the V group, and 1.55 in the VB group. The level of MMP-2 activity was similar in the VB and V groups. CONCLUSIONS: The level of MMP-9 activity increased in association with VB. The role of MMP-9 in the pathogenesis of VB should be verified.
Esophageal and Gastric Varices ; Hemorrhage ; Humans ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; Ulcer ; Varicose Veins

PURPOSE: We investigated the extent of adenosine A1 agonist-induced expression and regulation of matrix metalloproteinase 2 (MMP-2) synthesis in human trabecular meshwork cells (HTMC). METHODS: Primary HTMC cultures were exposed to 0.1 or 1.0 µM N6-cyclohexyladenosine (CHA) for 2 h in the presence or absence of an inhibitor thereof, 8-cyclopentyl-1,3-dimethylxanthine (CPT). The expression level of mRNA encoding MMP-2 was assessed via reverse transcription-polymerase chain reaction, and the levels of tissue inhibitor of metalloproteinase 2 (TIMP2) and membrane-type-1 MMP (MT1-MMP) measured by Western blotting. The permeability of the HTMC monolayer was assessed with the aid of carboxyfluorescein. RESULTS: CHA at 1.0 µM increased the permeability of the HTMC monolayer (p = 0.003) and CHA at both 0.1 and 1.0 µM significantly increased MMP-2 mRNA expression, which was inhibited by co-exposure to CPT (all p < 0.05). CHA increased MMP-2 activity, decreased that of TIMP2, and increased that of MT1-MMP (all p < 0.05). CONCLUSIONS: CHA increased the permeability of the HTMC monolayer and increased MMP-2 activity, decreased TIMP2 activity, and increased MT1-MMP activity. Thus, regulation of TIMP2 and MT1-MMP expression may be involved in the adenosine A1 agonist-induced increase in MMP-2 activity.
Adenosine ; Blotting, Western ; Humans ; Matrix Metalloproteinase 14 ; Matrix Metalloproteinase 2 ; Permeability ; RNA, Messenger ; Tissue Inhibitor of Metalloproteinase-2 ; Trabecular Meshwork

OBJECTIVETo detect protein expression of MMPs and TIMPs in various salivary gland neoplasms and to investigate their roles in invasion and metastasis of the malignant salivary gland tumors.

METHODSImmunohistochemistry and Gelatin zymography analyses for MMP-2, MMP-9, MT1-MMP, TIMP-1 and TIMP-2 were performed in 26 malignant and 28 benign salivary gland tumors.

RESULTSThe expression of MMP-2 and MMP-9 was significantly higher in carcinomas than in adenomas (P < 0.05). The MMP-2/TIMP-1 and MMP-2/TIMP-2 was also significantly higher in carcinomas than in adenomas (P < 0.05). There was a cooperated effect among MMP-2, MT1-MMP and TIMP-2. The expression of active MMP-2, proMMP-9 and active MMP-9 was significantly higher in malignant tumors than in benign tumors (P < 0.05).

CONCLUSIONMMP-2 and MMP-9 may play important roles in invasion of malignant salivary gland tumors. A disturbed balance between MMP-2, MMP-9, TIMP-1 and TIMP-2 in malignant salivary gland tumors was detected. It was the absolute increase of MMP-2 and MMP-9 to induce the unbalance.

Enzyme Precursors ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; Salivary Gland Neoplasms ; Salivary Glands ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

The high performance liquid chromatography (HPLC) and enzyme-linked immunoassay (ELISA) were used to investigate the changes of collagen and matrix metalloproteinase-1 (MMP-1) in liver, lung and kidney during growth process of mice. The mice from 0 to 18 weeks were used as the research objects. The contents and proportions of hydroxyproline (Hyp), which were used to calculate the collagen contents, in liver, lung and kidney of different weeks were analyzed with HPLC. The contents and activity of MMP-1 in liver, lung and kidney of different weeks were analyzed with ELISA. The results showed that the collagen contents in liver, lung, and kidney were different (Lung(COL)>Kidney(COL)>Liver(COL)), and they all increased first and then decreased with weeks. The collagen contents in liver, lung, and kidney reached the highest level in the ninth (5.52 ng/mg), sixth (54.10 ng/mg) and ninth (19.20 ng/mg) week, respectively. Then it declined slowly from 9 to 18 weeks. The result of ELISA showed that the MMP-1 contents in liver, lung and kidney decreased first and then increased with weeks, and the trend of MMP-1 activity was opposite. It indicated that the increase of collagen contents in the tissues will inhibit the secretion of MMP-1.
Animals ; Collagen ; Kidney ; Liver ; Lung ; Matrix Metalloproteinase 1 ; Matrix Metalloproteinase 2 ; Mice

BACKGROUND : Tumor invasion and metastasis are the major causes of morbidity and death for cancer patients. Metastasis is a complex multistep process in which tumor cells must pass through supporting structures. Proteolytic degradation of the structures is an important part of this process, and matrix metalloproteinase (MMP) has been implicated. The activation and the enzymatic activity of MMP is regulated by the tissue inhibitor metalloproteinase (TIMP). Three distinct TIMP molecules have been isolated. Very little is known about the role of TIMP-2 in tumors. The purpose of this study was to examine the expression of TIMP-2 in human colorectal carcinomas. METHODS : The paraffin blocks of 33 colorectal carcinomas were recalled and immunostained with monoclonal antibodies specific for TIMP-2. The rate of stain was estimated, and the relationships between the expression and the stage, the differentiation, and the recurrence were assessed. RESULTS : The expression of TIMP-2 in tumor tissues increased with increasing Dukes's stage (p<0.05) and recurred cases (p<0.05). CONCLUSIONS : Our results suggest that increased expression of TIMP-2 may be useful as a marker of biologic aggressiveness.
Antibodies, Monoclonal ; Colorectal Neoplasms* ; Humans* ; Matrix Metalloproteinase 2* ; Neoplasm Metastasis ; Paraffin ; Recurrence ; Tissue Inhibitor of Metalloproteinase-2

OBJECTIVETo explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.

METHODSFresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.

RESULTSAmeloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).

CONCLUSIONThe xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.

Ameloblastoma ; Animals ; Chickens ; Chorioallantoic Membrane ; Heterografts ; Humans ; Matrix Metalloproteinase 2 ; Tissue Inhibitor of Metalloproteinase-2 ; Transfection

OBJECTIVE: This study was performed to investigate the influence of hepatocyte growth factor (HGF) on matrix metalloproteinase (MMP), which are related in the lysis process of tissue during the invasion of trophoblasts. METHOD: HT cell line was treated with recombinant HGF (rHGF) of different concentration (0, 10, 50 and 100 ng/mL) and was cultured for 24 hours to check the changes in the expression of MMP-2 and MMP-9. Also, HT cell line was treated with recombinant HGF 50 ng/mL and was cultured for 24, 36, 48, and 72 hours to check the changes in the expression of MMPs according to the different time span. Total RNA were extracted from each cultured sample and RT-PCR and Western blotting were used to analyze the expression of MMP-2 and MMP-9. RESULTS: MMP-2 mRNA expression with treated rHGF showed increase of 2, 2.5 and 2.2 times with the increase of concentration level of 10, 50 and 100 ng/mL accordingly, while MMP-2 protein expression were increased 1.4 and 1.5 times in 50 ng/mL and 100 ng/mL of rHGF respectively compared with that of normal control. MMP-9 mRNA showed no significant changes in its expression with all different levels of concentration, while MMP-9 protein showed 1.5 times increase with 10 ng/mL rHGF but 0.4 times decrease with 100 ng/mL. MMP-2 mRNA expression treated with recombinat HGF were increased 1.6 times with 24 hour culture and 2.3 times with 36 hour culture. MMP-2 protein showed 1.9 times increase only for the case of 24 hour culture. MMP-9 mRNA expression of recombinant HGF-treated groups was decreased 0.7 times compared with that of control group in 36 hours. MMP-9 protein expression were increased by 1.2, 1.6 and 1.9 times as culture time increase to 36, 48, and 72 hours accordingly, compared with that of normal control. CONCLUSION: This result suggests that the HGF might partially regulate the invasion of trophoblasts through MMP-2 and MMP-9.
Blotting, Western ; Cell Line* ; Hepatocyte Growth Factor* ; Hepatocytes* ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; RNA ; RNA, Messenger ; Trophoblasts

OBJECTIVE: This study was performed to investigate the influence of hepatocyte growth factor (HGF) on matrix metalloproteinase (MMP), which are related in the lysis process of tissue during the invasion of trophoblasts. METHOD: HT cell line was treated with recombinant HGF (rHGF) of different concentration (0, 10, 50 and 100 ng/mL) and was cultured for 24 hours to check the changes in the expression of MMP-2 and MMP-9. Also, HT cell line was treated with recombinant HGF 50 ng/mL and was cultured for 24, 36, 48, and 72 hours to check the changes in the expression of MMPs according to the different time span. Total RNA were extracted from each cultured sample and RT-PCR and Western blotting were used to analyze the expression of MMP-2 and MMP-9. RESULTS: MMP-2 mRNA expression with treated rHGF showed increase of 2, 2.5 and 2.2 times with the increase of concentration level of 10, 50 and 100 ng/mL accordingly, while MMP-2 protein expression were increased 1.4 and 1.5 times in 50 ng/mL and 100 ng/mL of rHGF respectively compared with that of normal control. MMP-9 mRNA showed no significant changes in its expression with all different levels of concentration, while MMP-9 protein showed 1.5 times increase with 10 ng/mL rHGF but 0.4 times decrease with 100 ng/mL. MMP-2 mRNA expression treated with recombinat HGF were increased 1.6 times with 24 hour culture and 2.3 times with 36 hour culture. MMP-2 protein showed 1.9 times increase only for the case of 24 hour culture. MMP-9 mRNA expression of recombinant HGF-treated groups was decreased 0.7 times compared with that of control group in 36 hours. MMP-9 protein expression were increased by 1.2, 1.6 and 1.9 times as culture time increase to 36, 48, and 72 hours accordingly, compared with that of normal control. CONCLUSION: This result suggests that the HGF might partially regulate the invasion of trophoblasts through MMP-2 and MMP-9.
Blotting, Western ; Cell Line* ; Hepatocyte Growth Factor* ; Hepatocytes* ; Matrix Metalloproteinase 2 ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; RNA ; RNA, Messenger ; Trophoblasts

PURPOSE: Matrix metalloproteinases (MMPs) are endogenous peptidases that are capable of degrading various components of the basement membranes. To evaluate the clinical significance of the expressions of MMPs in renal cell carcinomas (RCCs), the MMPs' expression in RCCs and non- neoplastic kidney tissues was examined to evaluate the clinical significance of the expressions of MMPs in renal cell carcinomas (RCCs). MATERIALS AND METHODS: Twenty-two patients with RCCs (the RCC group), and eleven patients with non-neoplastic kidneys (the control group), were enrolled in this study between November 2002 and November 2003. The MMP-2 and MMP-9 activities were estimated using gelatin zymography, and they were quantified using a laser densitometer. The results were compared with the clinicopathological characteristics. RESULTS: The expression of MMP-9 was significantly elevated in the RCC group compared with the control group (p<0.01). There was no difference in MMP-2 activity between the RCC group and the control group (p>0.05). The levels of MMP-9 expression in the RCC patients with a large tumor (>4cm) or vascular invasion were significantly higher than that in the patients without these clinical manifestations (p<0.01). There were also significant differences in the expression of MMP-9 among the T stages (p<0.01). CONCLUSIONS: The present study shows a close relationship between the expression of MMP-9 and the tumor size and tumor stage in RCC. MMP-9 may be used as a prognostic marker and for the development of a novel treatment modality for RCC.
Basement Membrane ; Carcinoma, Renal Cell* ; Gelatin ; Humans ; Kidney ; Matrix Metalloproteinase 2* ; Matrix Metalloproteinase 9* ; Matrix Metalloproteinases ; Peptide Hydrolases

OBJECTIVETo investigate the relationship between matrix metalloproteinase-2 (MMP-2)activity and cell proliferation, growth and invasion of ameloblastoma.

METHODSThe cells and xenograft of ameloblastoma were treated with MMP-2 inhibitor Ro31-9790 and the effects of Ro31-9790 on the cell proliferation and growth of ameloblastoma were observed. Primary culture in vitro, subcapsular kidney xenograft in vivo, MTT assay, flow cytometry, neoplastic volume measurement and histochemistry were employed to study the effects of cell proliferation and growth produced by Ro31-9790.

RESULTSThere was no significant different in cell proliferation at same interval among several groups (P > 0.05). The ratio of G0/G1 stage, G2/M stage and apoptotic cells didn't increase following increased Ro31-9790, and the ratio of S stage cells also didn't reduce following increased Ro31-9790. The tumor volume and its increase in treatment group were significant less than those in control group.

CONCLUSIONRo31-9790 does not influence proliferation of ameloblastoma cells in vitro, but it can effectively inhibit the ameloblastoma growth in vivo. MMP-2 activity has no relationship to proliferation of ameloblastoma cells, but it can contribute to the ameloblastoma growth and may be a reason of invasion in ameloblastoma.

Ameloblastoma ; Cell Proliferation ; Humans ; Matrix Metalloproteinase 2 ; Neoplasm Invasiveness