BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the remodelling of extracellular matrix (ECM), including basement membrane. ECM remodelling is associated with pathological processes, including hepatic fibrosis, tumor invasion and metastasis. Tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 were known to inhibit MMP-9 and MMP-2, respectively. In the present study, we examined the expression of TIMP-1 and TIMP-2 in surgical specimen pairs of hepatocellular carcinoma and nontumoral liver and the correlation between their expression and clinicopathological characteristics. METHODS: The localization of both transcripts and protein of TIMP-1 and TIMP-2 was studied by using in situ hybridization and immunohistochemistry. RESULTS: TIMP-1 and TIMP-2 mRNA transcripts were found in tumor cells, hepatocyte, sinusoidal cells, endothelial cells and stromal cells. Signal intensity of TIMP-1 was stronger than that of TIMP-2. The results of immunohistochemical stainings were concordant with those obtained by in situ hybridization. Expression of TIMP-1 and TIMP-2 was observed in tumorous tissue, in nontumorous tissue and in the portions of the tumors adjacent to the capsules. However, a clear difference in TIMP-1 and TIMP-2 mRNA expression was not observed among the three tissue types. The intensity of TIMP-2 expression was generally weaker than that of TIMP-1, and the intensity of TIMP-1 and TIMP-2 mRNA expression did not correlate with variable clinicopathological characteristics. CONCLUSION: TIMPs was expressed in tumor cells and many cell types of the nontumoral liver. Further investigations for TIMPs' unknown functional role are needed.
Adult ; Aged ; Carcinoma, Hepatocellular/pathology ; Carcinoma, Hepatocellular/metabolism* ; Female ; Human ; Immunohistochemistry ; Liver Neoplasms/pathology ; Liver Neoplasms/metabolism* ; Male ; Middle Age ; RNA, Messenger/analysis ; Tissue Inhibitor-of Metalloproteinase-2/physiology ; Tissue Inhibitor-of Metalloproteinase-2/genetics* ; Tissue Inhibitor-of Metalloproteinase-2/analysis ; Tissue-Inhibitor of Metalloproteinase-1/physiology ; Tissue-Inhibitor of Metalloproteinase-1/genetics* ; Tissue-Inhibitor of Metalloproteinase-1/analysis

Female ; Hepatitis, Chronic ; metabolism ; Humans ; Immunohistochemistry ; Liver Cirrhosis ; metabolism ; Male ; Matrix Metalloproteinase 2 ; analysis ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

OBJECTIVES: To investigate expressions of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in squamous cell carcinoma of the tonsil and to correlate expression profiles with clinicopathological characteristics. METHODS: Paraffin blocks were obtained from 45 tonsil squamous cell carcinoma (SCC) patients, who underwent surgery as an initial treatment between 1994 and 2004, and from 20 normal controls. Expressions of MMP-2, MMP-9, MMP-13, TIMP-1, and TIMP-2 were investigated immunohistochemically. RESULTS: The expressions of MMPs (except MMP-2) and TIMPs were found to be significantly different in tonsil SCC and normal control tissues. Furthermore, MMP-13 expression was found to be correlated with tumor invasion (P=0.05), and the expressions of MMP-9 and TIMP-1 with nodal metastasis (P=0.048, 0.031). No relation was found between MMP or TIMP expression and recurrence. However, MMP-9 expression was found to be significantly associated with 5-year survival in tonsil SCC patients by multivariate analysis (hazard ratio, 3.853; P=0.013). CONCLUSION: Significant overexpressions of multiple MMPs and TIMPs were found in tonsil SCC tissues. Furthermore, our findings suggest that MMP-9 expression might be a useful prognostic factor.
Carcinoma, Squamous Cell ; Humans ; Matrix Metalloproteinases ; Metalloproteases ; Multivariate Analysis ; Neoplasm Metastasis ; Palatine Tonsil ; Paraffin ; Prognosis ; Recurrence ; Tissue Inhibitor of Metalloproteinase-1 ; Tissue Inhibitor of Metalloproteinase-2

OBJECTIVETo study the effects of hypoxia and hyperoxia on the expression and activity regulation of matrix metalloproteinase-2 (MMP-2) of the hepatic stellate cell (HSC).

METHODSThe expression of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC under hypoxic or hyperoxic conditions were detected with immunocytochemistry (LSAB method), the contents of MMP-2, TIMP-2 in culture supernatant with ELISA and the activity of MMP-2 in supernatant with zymography.

RESULTS(1) In the situation of hypoxia for 12 h, the expression of MMP-2 increased (hypoxia group positive indexes: 5.7 +/- 2.0; control: 3.2 +/- 1.0; P < 0.01), while TIMP-2 decreased (hypoxia group positive indexes: 2.5 +/- 0.7; control: 3.6 +/- 1.0; P < 0.05) in HSC, and the activity of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922; control: 17.277 +/- 7.424; P < 0.01). At the different time courses of hypoxia, the change of expression and activity of MMP-2 was most notable at 6 h. (2) In the situation of hyperoxia for 12 h, the protein contents of MMP-2, TIMP-2 in supernatant were both higher than those of the control, especially the TIMP-2 (hyperoxia group A(450): 0.050 +/- 0.014; control: 0.022 +/- 0.010; P < 0.01), and so was the activity of MMP-2 (hyperoxia group total A: 5.252 +/- 0.771; control: 4.304 +/- 1.083; P < 0.05). The expression of MT1-MMP was also increased.

CONCLUSIONSThe HSC is sensitive to the oxygen. Hypoxia accelerates the expression of MMP-2 and the effect is more marked at the early stage. Hyperoxia increases the activity of MMP-2.

Animals ; Cell Hypoxia ; physiology ; Hyperoxia ; enzymology ; Liver ; cytology ; enzymology ; Male ; Matrix Metalloproteinase 2 ; analysis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

OBJECTIVETo investigate the anticancer and anti-metastatic effect of Ajuga decumbens extraction (HBG) on breast cancer and to clarify the effect of HBG on MMPs and TIMPs.

METHODThe antitumor and antimetastic effect of HBG was determined using orthotopic 4T1 breast cancer mouse model. Western blot analysis was employed to detect the expression of associated proteins in breast cancer metastasis.

RESULTAdministration with 50-200 mg x kg(-1) doses of HBG significantly reduced the tumor weight, tumor volume and numbers of lung tumor nodules in a dose-dependent manner. Tumor metastasis correlated proteins were altered following HBG treatment, MMP-2 and MMP-9 were down-regulated while TIMP-1 and TIMP-2 were up-regulated.

CONCLUSIONHBG showed anticancer and antimetastatic effect towards breast cancer through regulating the expression of MMPs and TIMPs. These data sustain our contention that HBG might be used as a potential therapeutic agent.

Ajuga ; Animals ; Female ; Mammary Neoplasms, Experimental ; chemistry ; drug therapy ; pathology ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Metalloproteases ; analysis ; Mice ; Mice, Inbred BALB C ; Neoplasm Invasiveness ; Neoplasm Metastasis ; prevention & control ; Phytotherapy ; Plant Extracts ; therapeutic use ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; Tissue Inhibitor of Metalloproteinases ; analysis

OBJECTIVETo observe the changes in matrix metalloproteinase-2,7 (MMP-2,7) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in deep partial thickness burn during the process of wound healing, and the effects of bFGF on wound healing.

METHODSThe rats inflicted by 30% TBSA deep partial thickness burn were randomly divided into simple scald and bFGF treatment groups. Biopsies from wound skin were harvested at 3 and 6PBHs and 1, 3, 7, 14 PBDs for the detection of the epithelialization rate and collagen content. The above indices were also detected in the skin of another 6 normal rats as normal control.

RESULTS(1) The epithelialization rate in bFGF treatment group was higher than that in simple scald group during 3PBH to 14 PBD. (2) The collagen contents in both bFGF treatment group and simple scald group were continually decreased during 3 PBH to 3 PBD, and increased from 7 to 14 PBD, but still lower than that in normal control (P < 0.05). (3) The expression of MMP-2,7 and TIMP-2 in simple scald group enhanced from 1 to 14 PBD, and peaked on 7 PBD. (4) The expression of MMP-2,7 in bFGF treatment group was similar to that in simple scald group from 3 to 6 PBH, while the expressions of MMP-2,7 and TIMP-2 was higher than those in simple scald group from 1 to 14 PBD.

CONCLUSIONThe collagen deposition would be affected by the activities of extracellular matrix in scald wound in rats. Changes in MMP-2,7 and TIMP-2 expressions were an important process of wound repair, which was closely related to the acceleration of wound healing by the application of bFGF.

Animals ; Burns ; metabolism ; Collagen ; analysis ; Epithelium ; physiology ; Fibroblast Growth Factor 2 ; pharmacology ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 7 ; analysis ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; Wound Healing

Increased production of matrix metalloproteinases (MMPs) has been associated with increases in invasive and metastatic potential in many types of human carcinoma. Tissue inhibitors of metalloproteinase (TIMP)-1 inhibits most interstitial collagenases and MMP-9. TIMP-2 binds specifically and noncovalently to the pro-form of MMP-2 and inhibits its enzyme activity. In this study, we examined TIMP-1 and TIMP-2 expressions in relation to clinicopathological variables in colorectal carcinoma with in situ hybridization and immunohistochemistry. TIMP-1 and TIMP-2 expressions were localized overwhelmingly to pericancer stromal cells, while malignant and normal mucosal cells were weak or negative. Strong stromal TIMP-1 immunoreactivity correlated with Dukes' stage (p=0.022), status of lymph node metastasis (p=0.044) and poor survival (p= 0.005). The degree of immunohistochemical staining of TIMP-2 did not correlate with all clinicopathological variables. The correlation between enhanced TIMP-1 expression and advanced stage and poor survival suggest a growth promoting activity of TIMP-1 in colorectal carcinoma.
Adenocarcinoma/pathology ; Adenocarcinoma/mortality ; Adenocarcinoma/enzymology* ; Adult ; Aged ; Aged, 80 and over ; Antibodies ; Collagenases/immunology ; Collagenases/genetics* ; Collagenases/analysis ; Colorectal Neoplasms/pathology ; Colorectal Neoplasms/mortality ; Colorectal Neoplasms/enzymology* ; DNA Probes ; Female ; Gelatinase A ; Gelatinase B ; Gelatinases/immunology ; Gelatinases/genetics* ; Gelatinases/analysis ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Human ; In Situ Hybridization ; Male ; Metalloendopeptidases/immunology ; Metalloendopeptidases/genetics* ; Metalloendopeptidases/analysis ; Middle Age ; Predictive Value of Tests ; RNA, Messenger/analysis ; Stromal Cells/pathology ; Stromal Cells/enzymology ; Survival Analysis ; Tissue Inhibitor-of Metalloproteinase-2/immunology ; Tissue Inhibitor-of Metalloproteinase-2/genetics* ; Tissue Inhibitor-of Metalloproteinase-2/analysis ; Tissue-Inhibitor of Metalloproteinase-1/immunology ; Tissue-Inhibitor of Metalloproteinase-1/genetics* ; Tissue-Inhibitor of Metalloproteinase-1/analysis

BACKGROUNDE-cadherin, beta-catenin, cathepsin D, matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 are all invasion-related proteins. The expression patterns of these proteins in invasive ductal breast carcinomas, and their associations with known clinicopathological parameters, tumor recurrence and expressions of estrogen receptor (ER), progesterone receptor (PR), PS2 and c-erbB2 were not well studied in Chinese patients.

METHODSIn a set of 94 invasive ductal breast carcinomas, protein expressions of these molecular markers were investigated by immunohistochemistry, and their associations with known clinicopathological parameters, tumor recurrence and expressions of ER, PR, PS2 and c-erbB2 were also examined. In addition, the interrelationship between the expressions of these proteins were studied.

RESULTSPreserved membrane E-cadherin expression was associated with late tumor stage and tumor recurrence, whereas the reduced junctional beta-catenin associated with positive lymph node status and c-erbB2 overexpression. Positive staining of cathepsin D in tumor stromal cells displayed a significant association with late tumor stage. High expression of MMP-2 in cancer cells was associated with large tumor size and PR positive expression. TIMP-2 expression was positively associated with tumor recurrence. In addition, inter-relationship between the expressions of these biomarkers was also assessed. Cathepsin D staining in cancer cells was inversely correlated with its staining in stromal cells, and also inversely correlated with MMP-2 staining in tumor stromal cells. MMP-2 expression in stromal cells displayed an inverse correlation with TIMP-2 expression. MMP-9 expression displayed parallel associations with TIMP-1 and TIMP-2 expression.

CONCLUSIONEvaluation of E-cadherin, beta-catenin, cathepsin D, MMP-2 and TIMP-2 expression may be of some help in more accurately predicting the prognosis of invasive ductal breast carcinomas.

Adult ; Aged ; Breast Neoplasms ; chemistry ; pathology ; Cadherins ; analysis ; Carcinoma, Ductal, Breast ; chemistry ; pathology ; Cathepsin D ; analysis ; Female ; Humans ; Immunohistochemistry ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; Tissue Inhibitor of Metalloproteinase-2 ; analysis ; beta Catenin ; analysis

OBJECTIVETo study the role of changes of matrix metalloproteinase-2, 9 (MMP-2, 9) activity in the development of dimethylnitrosamine (DMN)-induced liver fibrosis in rats.

METHODSThe rat liver fibrosis model was established by peritoneal injection of DMN (at a dose of 10 mg/kg, 3 times a week, for 4 weeks). The dynamic changes of liver fibrosis were observed at different time points (1d, 2d, 3d, 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks). The MMP-2, 9 activity was measured by zymogram method. Liver ultrastructure was observed by electron microscope. The expressions of type IV collagen (CIV), laminin (LN), type I collagen (CI) and alpha-smooth muscle actin (alpha-SMA) were examined by immunohistochemistry. The tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) content was measured by Western blot method.

RESULTSThe MMP-2, 9 activity (gray value) significantly increased in the 2d and 3d DMN model rats (2d: normal/model group, MMP-2: 54.72+/-4.56/70.76+/-7.63; F = 16.27, P < 0.05; MMP-9: 25.72+/-4.29/51.76+/-15.33, F=13.38, P < 0.05). The positive staining area percentage of CIV in the sinusoidal walls decreased in the 2d, 3d and 1 weeks model rats (2d: normal/model group, 6.06+/-1.35/2.86+/-0.63, F=69.12, P < 0.05), but significantly increased in the 4w model rats (normal/model group, 6.06+/-1.35/8.04+/-1.50, F=14.42, P < 0.05). There was a remarkable negative correlation between the MMP-9 activity and expression of CIV in the sinusoidal walls (r = -0.729, P < 0.05). Positive expressions of LN and CI increased, and the strongest positive staining of them displayed in the 4w model rats. The formation of basement membrane was also observed in the 4 weeks model rats. Expression of TIMP-2 significantly increased in the late stage of fibrosis.

CONCLUSIONSThe increase of MMPs activity, especially MMP-9 which degrades the CIV normally distributed under the sinusoidal endothelium is the important factor in the formation of sinusoidal capillarization. The deposition and reconstitution of LN and new synthetic CIV, adding the deposition of CI constitute the high density basement membrane. The increase of TIMP-2 expression in the late stage of the fibrosis may be one of reasons why natural resolution of DMN-induced liver fibrosis is difficult.

Animals ; Collagen Type IV ; analysis ; Laminin ; analysis ; Liver ; chemistry ; ultrastructure ; Liver Cirrhosis, Experimental ; enzymology ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

OBJECTIVETo determine the molecular mechanisms involved in atrial fibrosis which occurs in patients with atrial fibrillation (AF) and to investigate their effects on the initiation and maintenance of AF.

METHODSThe right atrial tissue samples were taken from 73 patients with rheumatic heart disease who underwent heart valve replacement surgery. 34 patients had no history of AF (sinus rhythm group), 9 patients had paroxysmal AF and 30 patients had persistent AF. The mRNA content of collagen type I, collagen type III, MMP-2, TIMP-1, TIMP-2, TIMP-3 and TIMP-4 was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and normalized to beta-actin or GAPDH.

RESULTSCompared to sinus rhythm group, the mRNA of collagen type I and MMP-2 increased significantly in the persistent AF group (all, P < 0.01), followed by the paroxysmal AF group (all, P < 0.05). The mRNA of collagen type III was slightly higher in both AF groups than in the sinus rhythm group, but the differences were not statistically significant (P > 0.05). The mRNA of TIMP-1, TIMP-2 and TIMP-3 was down-regulated in the persistent AF group (all, P < 0.01, respectively), however, the trends of reduction did not reach statistical significance in the paroxysmal AF group (P > 0.05). The mRNA of TIMP-4 remained compatible in each group. The mRNA of collagen type I was significantly correlated with left atrial dimension (r = 0.336, P = 0.004) and AF duration (r = 0.339, P = 0.003). The mRNA of MMP-2 was significantly correlated with the mRNA of TIMP-2 (r = -0.326, P = 0.006), the mRNA of collagen type I (r = 0.322, P = 0.006), left atrial dimension (r = 0.300, P = 0.011) and AF duration (r = 0.300, P = 0.010).

CONCLUSIONThe increased level of collagen type I associated with selective downregulation of TIMP-2 and upregulation of MMP-2 gene expression in atrium could be one of the molecular mechanisms of atrial fibrosis during atrial fibrillation, which correlates with the initiation and maintenance of AF.

Adolescent ; Adult ; Atrial Fibrillation ; metabolism ; pathology ; Collagen Type I ; genetics ; Female ; Fibrosis ; Humans ; Male ; Matrix Metalloproteinase 2 ; genetics ; Middle Aged ; Myocardium ; pathology ; RNA, Messenger ; analysis ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Tissue Inhibitor of Metalloproteinases ; genetics