BACKGROUND: We investigated the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in malignant fibrous histiocytoma (MFH), and determined whether these could be useful as prognostic factors. METHODS: Among patients treated from 1993 to 2007, 30 cases of MFH were evaluated. Immunohistochemical staining was performed for MMP-2, MMP-9, TIMP-1, and TIMP-2 using paraffin wax-embedded blocks of MFH tissues. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot and zymography were performed using fresh tissues obtained from 17 of the 30 cases. The levels of MMP and TIMP expression were compared between the MFH and normal control groups, and between non-metastatic and metastatic MFH groups. RESULTS: Expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were higher in the MFH group than the control group by RT-PCR, Western blotting, and zymography. Immunohistochemical staining revealed that MMP-2 and MMP-9 protein expression was higher in the metastatic than in the non-metastatic group. The expression levels of MMP-2 and TIMP-1 were significantly higher in the metastatic than in the non-metastatic group (p < 0.05) by RT-PCR. By Western blot analysis, the expression levels of MMP-2, TIMP-1, and TIMP-2 were higher in the metastatic group (p < 0.05), but MMP-9 showed only a slight increase in the metastatic group compared with the non-metastatic group (p > 0.05). Finally, gelatin zymography analysis showed that the expression levels of the pro- and active forms of MMP-2 were significantly higher in the metastatic group (p < 0.05), but the expression of the pro- and active forms of MMP-9 showed a slight decrease in the metastatic group (p > 0.05). CONCLUSIONS: These results suggest that MMP-2, MMP-9, TIMP-1, and TIMP-2 may have important roles in the development and progression of MFH, and that the degree of expression of these metalloproteinases and their inhibitors, especially MMP-2, could be useful as prognostic factors related to metastasis in MFH.
Adult ; Aged ; Aged, 80 and over ; Female ; Histiocytoma, Malignant Fibrous/*metabolism ; Humans ; Immunohistochemistry ; Male ; Matrix Metalloproteinase 2/*biosynthesis ; Matrix Metalloproteinase 9/*biosynthesis ; Middle Aged ; Neoplasm Metastasis ; Prognosis ; Tissue Inhibitor of Metalloproteinase-1/*biosynthesis ; Tissue Inhibitor of Metalloproteinase-2/*biosynthesis

OBJECTIVETo investigate the effects of Herbal Compound 861 (Cpd 861) on collagen synthesis and degradation in rat mesangial cells exposed to high glucose.

METHODSThe third to fifth passage of rat mesangial cells were exposed to high glucose and Cpd 861 at a concentration of 0.25-4.00 g/L for 24, 48 and 72 h, respectively. Benazepril (10(-7)-10(-3) mmol/L) was selected as positive control. The methyl thiazolyl tetrazolium colorimetric assay was used to evaluate the effect of Cpd 861 on cell proliferation. After incubation with Cpd 861 at a concentration of 2.00 g/L for 48 h, the protein secretions of collagen type IV, matrix metallopeptidase 9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), transforming growth factor beta 1 (TGF-β1), and hepatocyte growth factor (HGF) were detected by enzyme-linked immunosorbent assay method. And rat mesangial cells were harvested to determine MMP-9, TIMP-1, TGF-β1 and HGF mRNA expression by reverse transcription polymerase chain reaction.

RESULTSCpd 861 inhibited cell proliferation induced by high glucose in a dose- and time-dependent manner. Compared with high glucose, collagen type IV production was decreased significantly by Cpd 861 (P<0.01). Cpd 861 increased the protein secretions and mRNA expressions of MMP-9 and HGF, whereas the protein secretions and mRNA expressions of TIMP-1 and TGF-β1 were reduced markedly (P<0.05). The ratio of MMP-9 to TIMP-1 was enhanced by Cpd 861 significantly. There was no significant difference in all above-mentioned effects between Cpd 861 (2.00 g/L) and benazepril (10(-5) mmol/L).

CONCLUSIONThe anti-glomerulosclerosis mechanisms of Cpd 861 were partly attributed to its effects of inhibiting mesangial cell proliferation, decreasing collagen synthesis and enhancing collagen degradation.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type IV ; biosynthesis ; secretion ; Drugs, Chinese Herbal ; pharmacology ; Fibrosis ; Glucose ; toxicity ; Hepatocyte Growth Factor ; secretion ; Matrix Metalloproteinase 9 ; metabolism ; Mesangial Cells ; cytology ; drug effects ; enzymology ; metabolism ; Polymerase Chain Reaction ; Proteolysis ; drug effects ; RNA, Messenger ; genetics ; metabolism ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Transforming Growth Factor beta1 ; secretion

OBJECTIVEThe aim of this study was to determine whether inhibition of cyclophilin A by lentivirus-mediated RNA interference (RNAi) could inhibit progression of atherosclerotic plaques and increase collagen production.

METHODSAtherosclerostic plaque model was induced by rapid perivascular carotid silicone collar placement in ApoE(-/-) mice. The recombinant CyPA-RNAi-Lentivirus (CyPA-RNAi-LV) or negative control-green fluorescent protein-Lentivirus (NC-GFP-LV) were constructed and transfected into right carotid plaques, respectively. Using the local injection method, ApoE(-/-) mice carotid artery plaque were intervened 10 min in the silicone collar placement with 10 µl (1.0 × 10⁸ TU/ml) lentivirus vector. The areas and CyPA expression of plaques were analyzed by morphological observation, real-time polymerase chain reaction (RT-PCR) and Western blot respectively.

RESULTSCyPA-RNAi-LV not only prevented plaques progression ((9 085 ± 671) µm² to (18 021 ± 1 901) µm²), but also decreased plaque lipid content ((28.9 ± 6.3)% to (17.8 ± 4.5)%), increased plaque collagen content ((24.2 ± 4.8)% to (35.1 ± 5.2)%) at 6 weeks after lentivirus transfection. The intima/media ratio (0.36 ± 0.11 vs. 0.65 ± 0.12, P < 0.05) and degree of lumen stenosis (intima/lumen ratios, 0.18 ± 0.02 vs. 0.33 ± 0.03, P < 0.05) were also significantly reduced by CyPA-RNAi-LV. Moreover, RT-PCR analysis revealed downregulated expressions of proinflammatory cytokines and matrix metalloproteinases (MMP-9 -17.5%) in the CyPA-RNAi-LV group.

CONCLUSIONLentivirus-mediated CyPA silencing by siRNA could inhibit plaques progression and reduce local inflammation through the anti-inflammatory effects in this model.

Animals ; Apolipoproteins E ; Cyclophilin A ; biosynthesis ; genetics ; Disease Progression ; Gene Silencing ; Genetic Vectors ; Inflammation ; Lentivirus ; Matrix Metalloproteinase 9 ; Matrix Metalloproteinases ; Mice ; Plaque, Atherosclerotic ; RNA Interference ; RNA, Small Interfering ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVEIn order to investigate the roles of metalloproteinase in inflammatory bone destruction in ankylosing spondylitis (AS), and analyze the mechanism of preventing inflammatory bone destruction of Bushen Qiangdu decoction (BSQDD) in AS cases. Comparisons were made on the expressions of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) by peripheral blood mononuclear cells (PBMC) between AS patients and healthy controls. The effect of BSQDD was investigated on the expression and of MMP-9 and TIMP-1 produced by PBMC in AS patients.

METHODSFrom March 2005 to March 2006, 30 active AS cases of Kidney-asthenia, Du-cold and blood-stasis syndrome were selected as patients group in the China-Japan Friendship Hospital. There are 27 male patients and 3 female patients. The age range is from 16 to 45, averaging (30.8 +/- 8.8) years. Disease duration is from 0.5 to 10 years. Cases received three-month BSQDD treatment were considered as the treatment group. Twenty healthy persons were included in the control group. Serum and PBMC were separated. The PBMC were stimulated by PHA and PMA, and the supernatant was collected. The mRNA expression of MMP-9 and TIMP-1 in PBMC was analyzed by RT-PCR. The content of MMP-9 and TIMP-1 in serum and culture supernatant of PBMC were detected by ELISA.

RESULTSCompared with health control group, the serum concentration of MMP-9 and TIMP-1 in patients group before treatment increased (P<0.01, P<0.05), but the level of MMP-9 and TIMP-1 in the serum of patients after treatment decreased compared with pre-treatment cases (P<0.05). Furthermore,compared with health control group, PBMC of patients group before treatment expressed higher levels of MMP-9 and TIMP-1 both on transcript level and at protein level (P<0.01, P<0.05), and the expression levels of MMP-9 and TIMP-1 in PBMC in patients after treatment both on transcript level and at protein level was down-regulated compared with pre-treatment (P<0.01, P<0.05).

CONCLUSIONPBMC of AS patients had a higher potential capacity for MMP-9 and TIMP-1. BSQDD possibly prevented inflammatory bone destruction of AS through inhibiting production of MMP-9 and TIMP-1 produced by PBMC.

Adolescent ; Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; blood ; genetics ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Retrospective Studies ; Spondylitis, Ankylosing ; blood ; drug therapy ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; blood ; genetics ; Young Adult

BACKGROUNDBlood coagulation factor VII (FVII) is physiologically synthesized in the liver and released into the blood. Binding of FVII to tissue factor (TF) is related to the metastatic potential of tumor cells, also a significant risk factor in the development of hepatic metastasis in patients with colorectal cancer (CRC). It has been found that some cancer cells can produce FVII extrahepatically. However, little is known about FVII and CRC. We therefore hypothesized that CRC cells may synthese FVII, leading to tumor invasion and metastasis.

METHODSWe detected the expression of FVII protein in 55 CRC specimens by immunohistochemical staining. The FVII mRNA in 45 of 55 CRC cases, 6 colon cancer cell lines and one hepatoma cell line was measured by real-time reverse transcription-PCR (RT-PCR). Transwell invasion assays were performed to evaluate the changes of cell migration and invasion of LoVo cancer cells in vitro. We further observed the likely effectors regulated by the TF/FVIIa complex Western blotting assay.

RESULTSExtrahepatic synthesis of FVII was detected in the cytoplasm of 32 (58.2%) CRC specimens by immunohistochemistry, but not in normal mucosa. Liver metastasis (P = 0.003) and TNM staging (P = 0.005) were significantly correlated with FVII antigen expression. The positive ratios in stages I, II, III and IV were 33.3%, 40.0%, 52.4% and 87.5%, respectively. The expression of FVII mRNA in CRC with hepatic metastasis was significantly higher than CRC without hepatic metastasis (5.33 ± 2.88 vs. 1.47 ± 0.51, P = 0.03). Ectopic FVIIa induced a slight increase (1.34-fold) in the number of migrating cells, which was inhibited by the specific TF antibody. The formation of TF/FVIIa complex resulted in a marked increase in the expression of matrix metalloproteinases (MMP)-2 (3.5-fold) and MMP-9 (4.7-fold) in a time-dependent and dose-dependent manner.

CONCLUSIONSExtrahepatic synthesis of FVII by CRC cells may promote tumor invasion and metastasis. MMPs, as downstream effectors of TF/FVIIa signaling, facilitate the development of metastasis in colon cancer.

Adult ; Aged ; Aged, 80 and over ; Cell Line, Tumor ; Cell Movement ; Colorectal Neoplasms ; metabolism ; pathology ; Factor VII ; analysis ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; secondary ; Male ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; RNA, Messenger ; analysis ; Thromboplastin ; physiology

OBJECTIVE: To investigate the expression and the relativity of RECK,RAGE and MMP-9 in nasopharyngeal carcinoma (NPC) tissues. METHOD: RECK, RAGE and MMP-9 were detected with immunohistochemical methods in 64 NPC specimens(30 cases with cervical lymph nodes metastasis, while the other 34 cases are not) and 30 specimens with chronic nasopharyngitis. RESULT: RECK hardly expressed in chronic nasopharyngitis and NPC tissues, However, strong expression of RECK could be seen in the surrounding inflammatory cells and matrix of cancer nests. The positive rates of RECK in NPC with and without cervical lymph nodes metastasis were 3.3% (1/30) and 11.8% (4/34), separately, There was no statistical significance between the groups (P > 0.05). The positive rates of RAGE in chronic nasopharyngitis and NPC tissues were 100% (30/30) and 70.3% (45/64) respectively, while MMP-9 were 46.7% (14/30) and 78.1% (50/64). The positive rates of RAGE in NPC with and without cervical lymph nodes metastasis were 86.7% (26/30) and 55.9% (19/34) separately, whereas MMP-9 were 90.0% (27/30) and 67.7% (23/34) respectively. They all showed statistical significance between the groups. MMP-9 had a negative correlation with RECK (r = -0.369, P < 0.05) and a positive correlation with RAGE in NPC (r = 0.471, P < 0.05). CONCLUSION The expression of RECK and RAGE in NPC were down-regulated, the expression of MMP-9 was up-regulated. While the expression of RECK in NPC with cervical lymph nodes metastasis was down-regulated, while RAGE and MMP-9 were up-regulated. The abnormal expression of the three genes may be related to the progression of NPC. RECK and RAGE may be involved in the invasion and metastasis of NPC by regulating the expression of MMP-9.
Adolescent ; Adult ; Aged ; Female ; GPI-Linked Proteins ; metabolism ; Glycation End Products, Advanced ; metabolism ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; Receptor for Advanced Glycation End Products ; metabolism ; Receptors, Cell Surface ; biosynthesis ; Young Adult

OBJECTIVETo investigate the effect of the sera of rabbits fed with Tongxinluo on the expression and secretion of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in U937 monocyte-derived macrophages.

METHODSAtherosclerosis was induced in rabbits by high-cholesterol feeding, and the serum was obtained from the rabbits after administration of the aqueous solution of Tongxinluo or simvastatin by gavage. U937 monocyte-derived macrophages were incubated with the sera at different concentrations for 24 hours, and the changes in MMP-9 and TIMP-1 gene expression and secretion were detected by RT-PCR and enzyme-linked immunosorbent assay, respectively.

RESULTSThe serum of rabbits fed with Tongxinluo concentration-dependently inhibited the expression and secretion of MMP-9 in U937 macrophages, but did not affect TIMP-1 expression or secretion.

CONCLUSIONTongxinluo may stabilize the atherosclerotic plaques by inhibiting the expression and secretion of MMP-9.

Animals ; Atherosclerosis ; blood ; etiology ; Cholesterol, Dietary ; administration & dosage ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Macrophages ; drug effects ; metabolism ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Serum ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; U937 Cells

UNLABELLEDTo investigate the expressions of fibrillin-1, elastin and matrix metalloproteinase-1 and -9 (MMP-1, 9) in chronic actinic dermatitis in elderly patients and explore the pathogenesis of the disease.

METHODSTwenty-three patients with chronic actinic dermatitis were examined for the expressions of fibrillin-1, elastin, MMP-1, and MMP-9 with immunohistochemistry in the skin lesions. Image analysis was carried out to measure MMP-1 and MMP-9 expressions semi-quantitatively.

RESULTSIn the skin lesions of patients with chronic actinic dermatitis, elastin expression was obviously reduced or absent in the papillary dermis. The elastic fibers were disorderly arranged in the reticular dermis with local aggregation in some regions. Obvious fibrillin-1 deposition was found in the reticular dermis. Increased expressions of MMP-1, but not that of MMP-9, was found in the skin lesions of the patients.

CONCLUSIONElastin and fibrillin-1 deposition can be found in the skin lesions in patients with chronic actinic dermatitis, suggesting the association of increased MMP-1 expression with the elastic tissue degeneration in the lesions. MMP-9 does not exhibit an obvious association with the pathogenesis of chronic actinic dermatitis in elderly patients.

Aged ; Elastin ; biosynthesis ; Female ; Fibrillin-1 ; Fibrillins ; Humans ; Immunohistochemistry ; Male ; Matrix Metalloproteinase 1 ; biosynthesis ; Matrix Metalloproteinase 9 ; biosynthesis ; Microfilament Proteins ; biosynthesis ; Middle Aged ; Photosensitivity Disorders ; etiology ; metabolism ; Sunlight ; adverse effects

OBJECTIVETo observe the expressions of metastasis-associated protein (S100A4) and matrix metalloproteinase 9 (MMP9) in human non-small cell lung cancer (NSCLC) and investigate their correlations to the infiltration, metastasis and prognosis of NSCLC.

METHODSThe expressions of S100A4 and MMP9 were detected in 41 NSCLC specimens and 6 normal lung tissue specimens using immunohistochemistry with SP method. Univariate and multivariate survival analysis were used to analyze the correlations of S100A4 and MMP9 to the clinicopathological characteristics and progrnosis of NSCLC.

RESULTSCompared with normal lung tissues, NSCLC showed significantly increased positivity for S100A4 and MMP9 expression (P<0.05); their expression were significantly higher in adenocarcinoma than in squamous cell carcinoma (P<0.01), and higher in metastatic NSCLC than in that without lymphatic metastasis (P<0.01). The positive expression rates of S100A4 and MMP9 were significantly higher in tumors in TNM stages III +IV than in stages II+I (P<0.05). S100A4 expression was positively correlated to tumor size (P<0.001), while MMP9 was inversely correlated to tumor differentiation (P<0.05). The expressions of S100A4 and MMP9 were both correlated to lymphatic metastasis, TNM stages and pathological types (P<0.05), and they also showed a mutual correlation (P<0.01). Univariate survival analysis confirmed the effects of histological types, lymphatic metastasis, clinical TNM stages and expressions of S100A4 and MMP9 on the survival time of NSCLC patients (P<0.001). Multivariate survival analysis identified clinical TNM stages and expressions of S100A4 and MMP9 as the independent factors affecting the prognosis of NSCLC (P<0.05).

CONCLUSIONThe expressions of S100A4 and MMP9 are up-regulated in NSCLC and have significant correlations to the clinical and biological behaviors of NSCLC. S100A4 and MMP9 status are independent prognostic predictors of NSCLC, and detection of their expressions may help evaluate the prognosis of NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; Middle Aged ; Neoplasm Invasiveness ; Prognosis ; S100 Calcium-Binding Protein A4 ; S100 Proteins ; biosynthesis

The present study aimed to investigate the effects of interleukin-1β (IL-1β) at different doses on collagen synthesis and decomposition in cultured cardiac fibroblasts from neonatal Sprague-Dawley rat. Cardiac fibroblasts were treated with IL-1β (0.01, 0.1, 1, 10, 100 ng/mL) for 24 h. Cell DNA synthesis was measured by (3)H-thymidine ((3)H-TdR) incorporation and collagen synthesis was measured by (3)H-proline ((3)H-Pro) incorporation. Matrix metalloproteinase (MMP) activity was measured by gelatinase zymography. MMP-2, MMP-9 protein expressions were measured by Western blot. mRNA expressions of MMP-2 and MMP-9 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Compared with that in the control group, the incorporation of (3)H-TdR and (3)H-Pro decreased in high-dose IL-1β groups (≥0.1 ng/mL) but not in low-dose IL-1β group (0.01 ng/mL). IL-1β significantly increased MMP-2 and MMP-9 activities. IL-1β (0.01-100 ng/mL) also dose-dependently increased the protein and mRNA expressions of MMP-2 and MMP-9 (P<0.05, P<0.01), respectively. These results suggest that IL-1β decreases collagen synthesis and MMP activities through transcriptional and posttranslational processes via degrading collagen in a dose-dependent way. Elevation of IL-1β is possibly involved in the process of ventricular remodeling after myocardial infarction, and the concentration of IL-1β is possibly a major factor which affects the extent of ventricular remodeling.
Animals ; Cells, Cultured ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; metabolism ; Interleukin-1beta ; pharmacology ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardial Infarction ; Myocardium ; cytology ; Rats ; Rats, Sprague-Dawley ; Ventricular Remodeling