mRNA Expression Levels Of MMP-9 Research Articles

Increased expression of collagen prolyl 4-hydroxylases in Chinese patients with hereditary gingival fibromatosis

Objectives Hereditary gingival fibromatosis (HGF) is characterized by excess accumulation of interstitial collagen. However, until now, there has been controversy about the mechanism of collagen accumulation in HGF gingivae. The present study aimed to clarify the pathogenic mechanisms potentially involved. Design Gingival fibroblasts from three Chinese HGF patients and three healthy subjects were cultured. Cell proliferation was assessed by MTT assay. The mRNA levels of type I collagen, MMP-1, MMP-3, TIMP-1, prolyl 4-hydroxylase (P4H)α(I), α(II), α(III) and P4Hβ were analyzed in gingival fibroblasts by RT-PCR. The protein production of type I collagen and P4H was examined respectively by ELISA and Western blot. Results In culture, HGF gingival fibroblasts showed similar growth characteristics to fibroblasts isolated from control gingivae. The mRNA and protein levels of type I collagen and P4Hα in HGF fibroblasts were higher than those in controls. There were no detected differences in mRNA expression levels of MMP-1, MMP-3, TIMP-1, P4Hα(II), α(III) and P4Hβ between HGF and control fibroblasts. Conclusions These data suggest that increased collagen post-translational modification by P4H may be one mechanism by which increased collagen accumulation occurs in some forms of HGF.

A single bout of exercise activates matrix metalloproteinase in human skeletal muscle

The aims of this study were 1) to characterize changes in matrix metalloproteinase (MMP), endostatin, and vascular endothelial growth factor (VEGF)-A expression in skeletal muscle in response to a single bout of exercise in humans; and 2) to determine if any exchange of endostatin and VEGF-A between circulation and the exercising leg is associated with a change in the tissue expression or plasma concentration of these factors. Ten healthy males performed 65 min of cycle exercise, and muscle biopsies were obtained from the vastus lateralis muscle at rest and immediately and 120 min after exercise. In the muscle biopsies, measurements of mRNA expression levels of MMP-2, MMP-9, MMP-14, and tissue inhibitor of metalloproteinase; VEGF and endostatin protein levels; and MMP activities were performed. Femoral arterial and venous concentrations of VEGF-A and endostatin were determined before, during, and 120 min after exercise. A single bout of exercise increased MMP-9 mRNA and activated MMP-9 protein in skeletal muscle. No measurable increase of endostatin was observed in the skeletal muscle or in plasma following exercise. A concurrent increase in skeletal muscle VEGF-A mRNA and protein levels was induced by exercise, with no signs of peripheral uptake from the circulation. However, a decrease in plasma VEGF-A concentration occurred following exercise. Thus 1) a single bout of exercise activated the MMP system without any resulting change in tissue endostatin protein levels, and 2) the increased VEGF-A protein levels are due to changes in the skeletal muscle tissue itself. Other mechanisms are responsible for the observed exercise-induced decrease in VEGF-A in plasma.

Lycopene prevents UVA-induced MDA formation and matrix metalloproteinases mRNA expression in rat skin

Objective To explore effects of lycopene on ultraviolet A （UVA）-induced malonic dialdehyde （MDA） formation and matrix metalloproteinases （MMPs）mRNA expression in rat skin. Methods Thirty Wistar rats were randomized into three groups comprising control group （C）, ultraviolet exposed group （UV）, and ultraviolet exposed and lycopene-treated group （UV＋L）. Rats of UV group and UV＋L group were irradiated with UVA radiation. A dose of 20 J/cm2 was given three times weekly for 12 weeks, yielding a total dose of 720 J/cm2. In rats of UV＋L group lycopene oil-resin was gavaged before irradiation. A dose of 5rag was given every day for 16 weeks. Lycopene-free oil-resin was gavaged accordingly in rats of C and UV groups. After UVA radiation, MMPs mRNA expression levels and MDA concentration were observed by semi-quantitative RT-PCR and modified thiobarbituric acid （TBA） microcolorimetric methods. Results MMP-1, MMP-2 mRNA expression levels and concentration of MDA in the UV＋L group were significantly lower than those in group UV （P〈0.01 ; P〈0.01）. MMP-9 mRNA expression levels in the two groups had no significant difference （P〉0.05）. Conclusions Lycopene can inhibit UVA-induced skin MMPs mRNA expression and MDA formation, which indicates that systemic use of lycopene may prevent UVA-induced photoaging in rats. Key words: Photoaging; Ultraviolet A; Lycopene; Matrix metalloproteinases

Expression of Snail, Slug and Sip1 in malignant mesothelioma effusions is associated with matrix metalloproteinase, but not with cadherin expression

Snail, Slug and Sip1 regulate cadherin and protease expression and mediate epithelial-mesenchymal transition in cancer. We analyzed the expression of cadherins and matrix metalloproteinases (MMP) and their transcriptional regulators in malignant mesothelioma (MM). One hundred and ten MM specimens (86 solid, 24 effusions) and 10 non-malignant effusions with reactive mesothelial cells (RMC) were analyzed for E-cadherin, N-cadherin and P-cadherin protein expression using immunhistochemistry. MM effusions were further analyzed for expression of Snail, Slug, Sip1, E-cadherin, MMP-2, MMP-9, MT1-MMP (MMP-14) and the MMP inhibitor TIMP-2, and for MMP-2 and MMP-9 activity using RT-PCR, Western blotting, immunhistochemistry and zymography. Results were analyzed for relationship with specimen type (biopsy versus effusion) and anatomic site (pleural versus peritoneal). E-cadherin, N-cadherin and P-cadherin expression was found in 69/110 (63%), 87/110 (79%) and 84/110 (76%) MM cases, respectively. Pleural and peritoneal MM showed comparable expression, but all three cadherins were upregulated in effusions compared to solid tumors (p<0.001). RMC were uniformly negative for E-cadherin and N-cadherin, and showed P-cadherin expression in 7/10 specimens. Immunohistochemistry localized MMP-2, MMP-9 and TIMP-2 to MM cells in 11/15, 14/15 and 8/15 effusions, respectively. RT-PCR showed direct association between MMP-2 mRNA expression level and the levels of MT1-MMP (p=0.027) and TIMP-2 (p=0.011). Snail protein expression showed positive association with MT1-MMP (p=0.016) and TIMP-2 (p=0.02) mRNA expression, but its expression was unrelated to MMP-2 and MMP-9 expression or activity. Snail, Slug and Sip1 levels did not show inverse association with E-cadherin levels. Our data show that E-cadherin and N-cadherin are selectively expressed in malignant mesothelial cells, and that P-cadherin and N-cadherin are expressed with similar frequency in MM. In agreement with our earlier data for ovarian carcinoma, cadherin expression is upregulated in effusions compared to solid lesions. The increased E-cadherin expression in effusions may be related to lack of negative regulation at the epigenetic level. The relationship between Snail and MMP in MM is uncertain at present.

The role of KiSS-1 and matrix metalloproteinase-9 in regulation of invasion of trophoblasts

To explore the role of metastasis-related gene KiSS-1 and matrix metalloproteinase (MMP)-9 in regulation of invasion of trophoblasts. RT-PCR and Western blotting were used to detect the MMP-9 and KiSS-1 expression levels in the placental tissues obtained from 90 cases of normal pregnant women undergoing artificial abortion, induction of labor with water bag or selective cesarean section among which 30 cases were in the first trimester, 30 in second-trimester and 30 cases of term pregnancy, and in the placental tissues of 40 cases of preeclampsia (15 cases of mild and 25 cases of severe preeclampsia) undergoing cesarean section, and tissues of 90 cases of hydatidiform mole, 9 cases of invasive mole and 8 cases of choriocarcinoma, all undergoing surgery. The expression levels of MMP-9 mRNA and protein were higher in first-trimester [A value 0.391 +/- 0.215, (36 +/- 7) microg/100 microg total protein] and then decreased gradually with the progress of gestation. The expression levels of MMP-9mRNA and protein in the term placental samples were significantly lower than those of first-trimester (both P < 0.01). The expression levels of KiSS-1mRNA and metastin in normal placenta increased along with the progress of gestation (both P < 0.01). The KISS-1 mRNA expression level and MMP-9 protein expression level in the placental tissue of preeclampsia were 0.09 +/- 0.06 (A value) and (9.6 +/- 4.3) microg/100 microg total protein respectively, both significantly lower than those of the term placenta (both P < 0.01). The expression levels of MMP-9 mRNA and protein in the tissues of gestational trophoblastic disease were significantly higher than those in the first-trimester placenta (both P < 0.01). The expression level of KISS-1 mRNA and metastin in the tissues of hyddatidiform mole and invasive mole were both significantly lower than those in the first trimester placenta (P < 0.05, P < 0.01). The expression of KiSS-1mRNA and metastin in the tissues of choriocarcinoma could not be detected. The expression of the invasion-related gene, MMP-9, is positively related with, while the invasion suppressor gene, KiSS-1, is negatively related with the invasive ability of trophoblasts. The interaction of these two genes plays an important role in regulation of the invasion of trophoblasts.

Tissue inhibitor of metalloproteinase 1 is an independent predictor of prognosis in patients with nonsmall cell lung carcinoma who undergo resection with curative intent

Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) play a role in the processes of extracellular matrix degradation. Changes in their expression levels have been observed in various tumor types, including lung carcinoma. However, their clinical significance and their prognostic importance in the progression of nonsmall cell lung carcinoma (NSCLC) remain to be specified. In this study, mRNA expression levels of MMP-1, MMP-9, TIMP-1, and TIMP-2 were evaluated in patients with resected NSCLC, and their associations with disease progression and prognosis were determined. Between June 1996 and December 1999, 116 patients underwent resection for NSCLC. Expression levels of MMPs and TIMPs were evaluated using Northern blot analysis in these NSCLC tissue samples and in 39 matched samples of normal lung tissue. MMP-1, MMP-9, and TIMP-1 expression levels were increased in tumor samples compared with matched, corresponding normal tissues. In contrast, TIMP-2 expression was decreased in tumor samples. MMP-1 tumor expression was correlated significantly with the evolution of lymph node status and tumor-lymph node-metastasis (TNM) stage. In contrast, MMP-9 tumor expression was correlated significantly with increased T stage. TIMP-1 overexpression was an independent predictor of worse survival in patients with NSCLC that was not associated with other prognosis factors, such as TNM stage. The overexpression of TIMP-1 was an independent prognostic marker in patients with NSCLC, and evaluating TIMP-1 may be important for identifying patients who are at greater risk of disease recurrence.

Resveratrol inhibits phorbol myristate acetate-induced matrix metalloproteinase-9 expression by inhibiting JNK and PKC delta signal transduction.

Proteolytic degradation of the extracellular matrix and tumor metastasis correlate with the expression of endopeptidases known as matrix metalloproteinases (MMPs). The expression of MMPs is regulated by cytokines and signal transduction pathways, including those activated by phorbol myristate acetate (PMA). We found that resveratrol, a phytoalexin present in grapes, significantly inhibits the PMA-induced increase in MMP-9 expression and activity. These effects of resveratrol are dose dependent and correlate with the suppression of MMP-9 mRNA expression levels. PMA caused about a 23-fold increase in MMP-9 promoter activity, which was suppressed by resveratrol. Transient transfection utilizing MMP-9 constructs, in which specific transcriptional factors were mutagenized, indicated that the effects of PMA and resveratrol were mediated via an activator protein-1 and nuclear factor-kappaB response element. Resveratrol inhibited PMA-mediated activation of c-Jun N-terminal kinase (JNK) and protein kinase C (PKC)-delta activation. Therefore, we conclude that the MMP-9 inhibition activity of resveratrol and its inhibition of JNK and PKC-delta may have a therapeutic potential, given that a novel means of controlling growth and invasiveness of tumors.

Effect of batimastat on Matrix metallopro-teinase-2 mRNA in rat hepatocellular carcinoma

AIM:To study the dynamic changes of matrix metalloproteinase2 mRNA in liver tissue during the experimental hepatocarcinogenesis and the effect of Batimastat on it. METHODS:Hepatocellular carcinoma was induced with the administration of diethy Initrosoamine(DENA) in rats. BB-94 was intraperitoneally injected to treat the experimental models. Reverse transcription polymerase chain reaction was used for quantitative analysis of MMP-2 mRNA expression level during the induction and therapy. RESULTS:The expression of MMP-2mRNA was increased throughout the hepatocarcinogenesis, and reach had its maximum plateau in the early hepatocarcinogenesis stage. BB-94 no effect on MMP-2mRNA expression. CONCLUSION:MMP-2mRNA expression is increasing during hepatocarcinogenesis, and BB-94 has no effect on MMP-2mRNA expression.

Relaxin enhances in-vitro invasiveness of breast cancer cell lines by up-regulation of matrix metalloproteases.

Until recently, relaxin (RLX) has been known predominantly for its effects on the reproductive system, where it induces remodelling of the extracellular matrix and up-regulation of matrix metalloproteases (MMPs). In solid cancers, tissue remodelling and MMP activation are essential for invasion and metastasis. We therefore investigated the effect of RLX on invasiveness and MMP expression of human breast cancer cell lines. Upon incubation with porcine RLX, the invasiveness of SK-BR3 cells was significantly increased. Similar effects could be achieved in MCF-7 cells, especially when RLX was combined with epidermal growth factor. Enhanced invasiveness was accompanied by up-regulation of MMP production and could be almost completely blocked by the MMP inhibitor FN 439. Zymography revealed increased secretion of MMP-2, -7 and -9, associated with up-regulated mRNA concentrations of MMP-2, -9, -13 and -14. mRNA expression levels of MMP-1, -3, -7, -8, -10, -11, -12 and of tissue inhibitors of metalloproteases-1, -2, -3 and -4 were either very low or not detectably influenced by RLX. Taken together, RLX enhances in-vitro invasiveness of breast cancer cell lines by induction of MMP expression. It remains to be clarified whether RLX might play a similar role in vivo and promote tumour progression.

Expression of Matrix Metalloproteinases-9 and Stromelysin-3 in Peripheral Blood in Patients with Lung Cancer

Background: Matrix metalloproteinases(MMP) are essential enzymes for tumor invasion and metastasis. Among the MMP family, elevated MMP-9 and stromelysin-3(STR-3) expression have been reported to be poor prognostic factors in lung cancer patients. To evaluate the possibility of a molecular diagnosis of lung cancer using peripheral blood, the mRNA expression level of MMP-9 and STR-3 was measured using a reverse transcriptase-polymerase chain reaction (RT-PCR) in patients with lung cancer. Methods : Ninety six patients(44 patients with lung cancer, 19 pulmonary infection, and 33 control) were included. To detect MMP-9 and STR-3 mRNA expression, RT-PCR was performed in peripheral blood mononuclear cells. ELISA was also used to measure the serum level of MMP-9. Results : MMP-9 was expressed more frequently in patients with a pulmonary infection(18/19, 94.7%) compared to lung cancer patients(26/44, 59.1%) or the controls (23/33, 69.7%) (p=0.018). On the other hand, STR-3 expression was observed more frequently in patients with lung cancer(37/44, 84.1%) compared to the lung infection patients(8/19, 42.1%) or control(20/33, 60.6%) (p=0.003). Among the lung cancer patients, MMP-9 was expressed more frequently when a tumor invaded the lymph nodes(17/24, 70.8%) compared to when a tumor did not(3/13, 23.1%) (p=0.005). The MMP-9 and STR-3 expression levels had no relationship with age, sex, tumor size, distant metastasis, or tumor histology. The serum MMP-9 concentration was not higher in lung cancer patients compared to patients with a pulmonary infection or the control subjects. Conclusion : STR-3 may be used as a diagnostic marker in the peripheral blood of lung cancer patients using RT-PCR. Further studies to evaluate the clinical significance of elevated STR-3 expression in lung cancer patients is recommended.