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Sample records for matrix metallo-proteinases mmp

  1. Differential expression analsis of matrix metallo-proteinase 20 (MMP20) in human esophageal cancer

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The aim of this study was to investigate the different expressions of MMP20 in the process of esophageal cancer. RT-PCR, immunohistochemistry and Western blot were employed to detect the mRNA expression and protein expression of MMP20. Compared with the corresponding normal esophagus tissues, results showed the common trend of the dramatical overexpression of MMP20 in the esopha-geal cancer tissues. The analysis of MMP20 expression in different clinical early stages of esophageal cancer by RT-PCR indicated that MMP20 was much more overexpressed in the esophageal cancer with invasion than in the early stages. This suggested that MMP20 might play an important role in the progression of esophageal cancer. It probably plays a critical role in tumor invasion.

  2. Functional proteomic of Matrix Metallo-proteinases (MMP) dedicated to the detection of active forms of MMP in complex proteome; Proteomique fonctionnelle dediee aux Metalloproteases Matricelles (MMPs): developpement d'une methode extremement sensible permettant la detection des formes actives des MMPs dans des proteomes complexes

    Energy Technology Data Exchange (ETDEWEB)

    David, A

    2007-07-15

    The Matrix Metallo-proteinases (M.M.P.) represent a family of Zinc dependent extracellular proteinases able to cleave collectively all the proteins constituting the extracellular matrix. Currently, 23 human M.M.P. have been identified and are characterized by their sequence in amino-acids and their highly conserved 3 D structure. These enzymes are expressed constitutively during the tissue remodeling process. Their over-expression in various diseases tightly related to inflammatory processes (arthritis, emphysema, cancer) described M.M.P. as choice therapeutic targets. However, as the tissue remodeling implicates modification of cellular contacts, M.M.P. appear currently as proteins involved in signalling pathways. Recent works demonstrating that M.M.P. are able to cleave substrates, which are different than proteins constituting the extracellular matrix, reinforce this vision. In order to identify the individual role and the protein expression level of M.M.P. in pathological context, we developed a new technique of functional proteomics dedicated to the detection of active forms of M.M.P. in tumour samples. This technique relied on the development of a new photoaffinity probe, based on the structure of a potent phosphinic inhibitor of M.M.P., allowing targeting and isolating active forms of M.M.P. by photoaffinity labelling. Furthermore, as the new developed probe incorporated a radioactive element, photoaffinity labelling permitted to radiolabel the targeted proteins. This probe demonstrated in vitro its remarkable ability to covalently modify the h M.M.P.-12, with a singular cross-linking yield, determined at 42 %, displaying an extremely sensitive detection (2.5 fmoles of h M.M.P.-12). When added to complex proteome, the photoaffinity probe presents the same sensibility of detection for the h M.M.P.-12 (5 fmoles); importantly, in this case, h M.M.P.-12 represents only 0.001 % of the totality of the proteins present in the sample. Moreover, this technique allows

  3. Effect of quercetin on expression of matrix metallo-proteinases and tissue inhibitor of matalloproteinase-1 in cultured rat hepatic stellate cells

    Institute of Scientific and Technical Information of China (English)

    康鲁平; 齐荔红; 张俊平; 周斌

    2003-01-01

    Objective: To study the effects of quercetin (QU) on matrix metallo-proteinases (MMPs), the tissue inhibitor of matalloproteinase-1 (TIMP-1), procollagen I and 2 proteoglycans (decorin and biglycan) mRNA expression in cultured rat hepatic stellate cell line HSC-T6 cells.Methods: Cells were treated with different concentrations of QU (12.5, 25, 50 μmol/L) or drug solvent (0.1 % Me2SO) for 24 h.mRNA expression was determined by reverse transcription polymerase chain reaction (RT-PCR).Results: QU (12.5 - 50 μmol/L) enhanced collagenase (rat MMP-13) and membrane type1-MMP (MMP-14) mRNA expression, decreased procollagen I mRNA expression in a concentration-dependent manner, but did not affect gelatinase-A (MMP-2), TIMP-1, decorin and biglycan expression.Conclusion: QU may decrease matrix deposition and increase matrix degradation, which might be beneficial to liver fibrosis.

  4. Matrix metalloproteinases-2, -9 and tissue inhibitor of metallo-proteinase-1 in lung cancer invasion and metastasis

    Institute of Scientific and Technical Information of China (English)

    MING Shu-hong; SUN Tie-ying; XIAO Wei; XU Xiao-mao

    2005-01-01

    @@ Lung cancer is a major cause of death from malignant disease due to its high incidence, malignant behavior and lack of major advancements in treatment strategies. The ability to invade tissues and establish colonies at remote sites is a defining characteristic of malignant neoplasms. Matrix metalloproteinases (MMPs) are zinc proteinases that degrade compounds of extracellular matrix (ECM). These enzymes have been implicated in tumour invasion and metastasis through degrading many extracellular matrix proteins especially MMP-2 and MMP-9, which are regarded as markers of tumour invasion and metastasis.1 The purpose of this study is to examine the role of MMP-9, MMP-2, tissue inhibitor of metalloproteinase-1 (TIMP-1) and MMP-9/TIMP-1 in tumour invasion and metastasis as well as the relationships between the mRNA expression of MMP-9 in white blood cells and MMP-9 levels in the plasma.

  5. Matrix metalloproteinase-2 and tissue inhibitor of metallo-proteinase-2 in colorectal carcinoma invasion and metastasis

    OpenAIRE

    Li, Bing-hui; zhao,Peng; Liu, Shi-Zheng; Yu, Yue-Ming; Han, Mei; Wen, Jin-kun

    2005-01-01

    AIM: To explore the relationship between matrix metallopr-oteinase-2 (MMP-2) and tissue inhibitor of metallopr-oteinase-2 (TIMP-2) in the development of colorectal carcinoma and to provide a valuable marker for clinical diagnosis.

  6. Effect of adding Matrix Metallo proteinase inhibitors on the degree of conversion of monomers to polymer an experimental bonding agent

    Directory of Open Access Journals (Sweden)

    Ghavam M.

    2009-11-01

    Full Text Available "nBackground and Aim: In spite of the achievements in the field of dental adhesives, we are facing challenges with dentine bonding resistance, strength and stability. According to recent studies the role of MMP inhibitors in association with bonding,s persistence and leakage reduction and restoration,s persistence is important. The aim of this study was to investigate the effect of doxycycline as a MMP inhibitor on the degree of conversion (DC of an experimental dental adhesive. "nMaterials and Methods: In this experimental study, a new dental adhesive blend was prepared by mixing doxycycline monohydrate (in concentrations of 0.0, 0.25, 0.5, and 1 wt.% with monomers. The monomers were composed of 12% Bis-GMA and 10% TMPTMA, 28% HEMA, and 50% Ethanol by weight for all groups. Comphorquinone and amines were chosen as photo initiator system. Degree of conversion of all adhesives was measured using FTIR spectroscopy. The results were analyzed using one-way ANOVA and Tukey post hoc tests. "nResults: The results showed that addition of 0.25, 0.5, and 1 weight percent doxycycline did not significantly reduce the DC of the adhesives compared to 0.0% control group (p>0.05%. "nConclusion: According to the results of this study, adding doxycycline to the adhesives did not adversely affect the DC.

  7. 肺间质纤维化大鼠肺组织基质金属蛋白酶及其组织抑制因子含量变化%Changes of lung tissue matrix metallo proteinase and its tissue inhibitor in pulmonary fibrosis rats

    Institute of Scientific and Technical Information of China (English)

    黄日红; 吴泰华; 张中和

    2001-01-01

    观察肺纤维化形成过程中基质金属蛋白酶(Matrix Metallo proteinas 简称MMPs)及其组织抑制因子(Tissue inhibitors of Metallo proteinases 简称TIMPs)含量的变化,探讨其在肺纤维化发病中的作用.将W istar大鼠60只,随机均分为对照组及模型组,气管内注入博莱霉素A5 5mg/kg,制备肺间质纤维化动物模型,观察注药后1、3、7、14及28d肺脏病理变化,利用酶谱法及免疫印记法分析肺组织MMP-2、MMP-9,TIMP-1的含量变化.结果显示各模型组pro-MMP-2、MMP -2、TIMP-1蛋白含量均较对照组增加,尤其7、14及28d组MMP-2较前明显增多.而MMP- 9变化不很明显.提示在肺纤维化形成过程中, pro-MMP-2、MMP-2 及TIMP-1都有所增高,MMP/TIMP比例失衡是最终导致肺间质纤维化形成的重要因素.

  8. Expression of matrix metallo-proteinase-28 in human nor-mal cytotrophoblast cells and a choriocarcinoma cell line,JEG-3

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    There are striking similiarities present between the behavior of invasive placental cells and that of invasive cancer cells. Matrix metalioproteinases (MMPs) are one of the most important mediators. MMP-28, the new member ofMMPs, was sequenced and identified recently. Expression of MMP-28 mRNA and protein in the cytotrophoblast cells anda choriocarcinoma cell line, JEG-3 cell, was conducted by zymography, RT-PCR and Northern blot. There is MMP-28mRNA expression in both the cytotrophoblast cells and JEG-3 cells by RT-PCR. The activity of MMP-28 in cytotrophoblast cells was significantly weaker than that in JEG-3 (P < 0.01) by zymography. Furthermore, mRNA expression of MMP-28 was significantly stronger (P < 0.001) in JEG-3than in human cytotrophoblast cells in a time-dependent way by Northern Blot. Our results suggest that MMP-28 may play a role in some of the tissue-remodeling events associated with normal pregnancy and tumor progression.``

  9. Expression of adhesion molecules, chemokines and matrix metallo- proteinases (MMPs) in viable and degenerating stage of Taenia solium metacestode in swine neurocysticercosis.

    Science.gov (United States)

    Singh, Satyendra K; Singh, Aloukick K; Prasad, Kashi N; Singh, Amrita; Singh, Avinash; Rai, Ravi P; Tripathi, Mukesh; Gupta, Rakesh K; Husain, Nuzhat

    2015-11-30

    Neurocysticercosis (NCC) is a parasitic infection of central nervous system (CNS). Expression of adhesion molecules, chemokines and matrix metalloproteinases (MMPs) were investigated on brain tissues surrounding viable (n=15) and degenerating cysticerci (n=15) of Taenia solium in swine by real-time RT-PCR and ELISA. Gelatin gel zymography was performed for MMPs activity. ICAM-1 (intercellular adhesion molecule-1), E-selectin, MIP-1α (macrophage inflammatory protein-1α), Eotaxin-1 and RANTES (regulated on activation, normal T cell expressed and secreted) were associated with degenerating cysticerci (cysts). However, VCAM-1 (vascular cell adhesion molecule-1), MCP-1 (monocyte chemotactic protein-1), MMP-2 and MMP-9 were associated with both viable and degenerating cysts. In conclusion, viable and degenerating cysticerci have different immune molecule profiles and role of these molecules in disease pathogenesis needs to be investigated.

  10. Phenotypic overlap between MMP-13 and the plasminogen activation system during wound healing in mice

    DEFF Research Database (Denmark)

    Juncker-Jensen, Anna; Lund, Leif R

    2011-01-01

    combined completely prevent wound healing. Both urokinase-type plasminogen activator and several matrix metallo proteinases (MMPs), such as MMP-3, -9 and -13, are expressed in the leading-edge keratinocytes of skin wounds, which may account for this phenotypic overlap between these classes of proteases....

  11. 血清白细胞介素-6、白细胞介素-17和基质金属蛋白酶9水平与一氧化碳中毒迟发性脑病的关系%The relationship between the level of interleukin-6, interleukin-17 matrix metallo proteinase 9 and delayed encephalopathy after acute carbon monoxide poisoning

    Institute of Scientific and Technical Information of China (English)

    陈艳荣; 李晓; 徐化利; 潘双双; 李丽

    2016-01-01

    Objective To explore the relationship between the level of IL-6,IL-17 and matrix metallo proteinase 9 (MMP9) with deleyed encephelopathy after aucte carbon monoxide poisoning metal protease (DEACMP).Methods Rarcdomly selected 58 healthy people and 71 (delayed encephalopathy after acute carbon monoxide poisoning,DEACMP) patients were taken as control and experimental group,respectively.Enzyme-linked immunosorbent assay(ELISA) was used to detect the expression of IL-6,IL-17 and MMP9.Results Compared with control,the expression of IL-6 and MMP9 were significantly increased in mild and severe DEACMP patients,while both IL-6 and MMP9 showed largest increase in severe DEACMP patients with statistical significanc (F =22.47,F =36.51,P < 0.05).There was no significant expression changes of IL-17 (F =19.62,P > 0.05).Pearson analysis showed that the expressions of IL-6 and MMP9 were positively correlated with DEACMP (r =0.58,P < 0.05).Conclusions The expression of IL-6 and MMP9 in serum of DEACMP patients were associated with the degree of carbon monoxide poisoning.%目的 探讨白细胞介素(IL)-6)、IL-17和基质金属蛋白酶9(MMP9)与一氧化碳中毒迟发性脑病(DEACMP)的关系.方法 随机抽取58例体检健康人群为对照组,抽取71例DEACMP患者为实验组,采用酶联免疫吸附试验(ELISA)检测DEACMP患者血清中IL-6、IL-17和MMP9的表达水平.结果 和对照组相比,轻重度DEACMP患者血清中IL-6和MMP9表达水平升高(F=22.47、F=36.51,P<0.05),其中在重度DEACMP患者血清中升高最显著;IL-17表达水平未有显著变化,差异无统计学意义(F=19.62,P>0.05).Pearson相关分析显示,DEACMP患者血清IL-6和MMP9浓度呈正相关(r=0.58,P<0.05).结论 DEACMP患者血清中IL-6和MMP9的表达水平与一氧化碳中毒的程度和预后密切相关.

  12. Association of genetic polymorphisms of angiotensin converting enzyme and matrix metallo proteinase-1 with idiopathic pulmonary fibrosis%血管紧张素转化酶及基质金属蛋白酶-1基因多态性与特发性肺纤维化的相关性

    Institute of Scientific and Technical Information of China (English)

    袁雅冬; 张颖; 孙长虹

    2013-01-01

    Objective To investigate the correlation between angiotensin converting enzyme(ACE)and matrix metallo proteinase (MMP)-1 gene polymorphisms and the risk of idiopathic pulmonary fibrosis (IPF) in a Han Chinese population from Hebei Province.Methods Eighty-four IPF patients and 100 controls were enrolled from the Second Hospital of Hebei Medical University.Polymerase chain reaction(PCR)and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) were used to detect ACE gene insertion/deletion (I/D) polymorphism and MMP-1 polymorphism respectively.The MMP-1 polymorphism was genotyped by DNA sequence analysis.Radioimmunoassay and ELISA were used to analyzed AngⅡ,MMP-1 and TIMP-1 levels in IPF patients and heahhy controls.Results There was a significant difference between the 2 groups in allele and genotype frequency distribution of ACE Insertion/Deletion polymorphism; frequency distribution of DD genotype and D allele of IPF patients were higher than those of the healthy control group (x2 =11.227,4.318,P < 0.05).There was no difference from different genders and ages on allele and genotype frequency distribution of ACE Insertion/Deletion polymorphism.(x2 =0.03-1.069,P > 0.05).There was no significant difference between the 2 groups in genotype and allele frequency distribution of MMP-1 1G/2G polymorphism(x2 =0.94 and 0.001,P > 0.05).The AngⅡ levels from DD genotype of both IPF patients and healthy controls were the highest,followed by the DI genotype and the Ⅱ genotype.The AngⅡ level of any genotype for ACE Insertion/Deletion polymorphism in the IPF group was higher than that in the healthy control group (all P < 0.05).The serum level of AngⅡ,MMP-1 and TIMP-1,as well as MMP-1/TIMP-1 ratio in the IPF group were higher than those in the healthy control group (all P <0.05).Conclusions The ACE polymorphism might be associated with IPF,and the serum level of AngⅡ was affected not only by the genetic background of ACE insertion

  13. Effect of Azadirachta indica (Neem) and Aloe vera as compared to subantimicrobial dose doxycycline on matrix metalloproteinases (MMP)-2 and MMP-9: An in-vitro study.

    Science.gov (United States)

    Kudalkar, Mithun D; Nayak, Aarati; Bhat, Kishore S; Nayak, Ranganath N

    2014-01-01

    A critical outcome of periodontal diseases is degradation of collagen in the periodontal tissues, by enzymes such as Matrix Metallo-Proteinases (MMPs). Doxycycline is known to down-regulate the activity of MMPs. Azadirachta indica (Neem) and Aloe vera are herbs known to have an anti-inflammatory effect. The present study was designed to evaluate the anti-inflammatory effect of Neem and Aloe vera by way of its inhibitory effect on MMP-2 and MMP-9 activity in cases of chronic periodontitis and compare it with doxcycline. A total of 30 subjects were enrolled in this study. Gingival tissue samples were obtained from patients diagnosed with the chronic periodontitis. The tissue extracts were treated with the said drug solutions and inhibition of MMP-2 and MMP-9 was analyzed. Enzymatic activity was detected by electrophoresis. The data was subjected to Student's paired t-test. The results showed that the activity of MMP-2 and MMP-9 was significantly decreased by the use of doxycycline, Neem and Aloe vera. A 53.5% reduction in the MMP-2 and 52.5% reduction in the MMP-9 activity was seen when samples were subjected to Neem treatment at the concentration of 1500 μg/ml. Tissues treated with Aloe vera in the concentration of 2000 μg/ml showed a 20.09% reduction in the MMP-2 and 20.4% reduction in the MMP-9 activity. Doxycycline in the concentration of 300 μg/ml, showed an 82.1% reduction in the MMP-2 and 82.6% reduction in the MMP-9 activity. The present study demonstrated an inhibitory effect of Neem and Aloe vera on MMP-2 and MMP-9, which are involved in the extracellular matrix degradation during periodontitis.

  14. Relationship of matrix metallo proteinase-9 dynamic expression at different time points with brain edema after cerebral hemorrhage%脑出血后不同时间点基质金属蛋白酶-9的动态变化及其与脑水肿的关系

    Institute of Scientific and Technical Information of China (English)

    崔巍; 谈颂; 许晓辉; 宋波; 许予明

    2010-01-01

    目的 探讨血清基质金属蛋白酶-9(MMP-9)水平在脑出血后不同时间点的动态变化在脑水肿形成过程中的作用.方法 采用放射免疫法检测90例符合纳入标准的急性脑出血患者及80例正常对照者24 h、72 h、7 d 和14 d时血清MMP-9水平,分析其在脑水肿形成过程中的作用.结果 脑水肿在入院72 h时达高峰,之后缓慢下降.在发病24 h 内患者血清MMP-9含量已明显升高,发病后72 h达高峰,与脑水肿高峰一致;7 d下降明显,与对照组比较均明显升高(P<0.01),第14天接近正常水平.血清MMP-9水平在脑出血后24 h、72 h与水肿体积、水肿比值呈正相关(P<0.01).结论 脑出血后血清MMP-9随着时间动态变化,MMP-9与脑水肿体积相关,与相对水肿体积相关性更强.%Objective To study the effect of dynamic expression at different time points of metallo proteinase-9 on brain edema after cerebral hemorrhage. Methods Serum levels of MMP-9 in 90 patients with brain hemorrhage and 80 normal patients were detected by radioimmunoassay on 24 h、72 h、7 d and 14 d.The relationship between levels of MMP-9 and brain edema after cerebral hemorrhage.Results Brain edema went up to the peak at 72 h and slowly decline after it. At 24 h, the serum level of MMP-9 were significantly higher, and reached the peak at 72 h according to the peak of cerebral edema, and then decreased significantly after 7 d. Cerebral hemorrhage group compared with the control group were significantly higher(P<0.01). The level of MMP-9 close to normal levels on 14 d. MMP-9 was positive correlated with edema volume and edema ratioat 24 h and 72 h (P=0.01).Conclusions Serum MMP-9 dynamic changes over time after cerebral hemorrhage. The level of MMP-9 was correlated with the brain edema volume and had more stronger correlation with relative size of brain edema. MMP-9 was correlated with inflammation after cerebral hemorrhage.

  15. Current Research into Interrelationship of Matrix Metallo Proteinases-9 and Kidney Diseases%基质金属蛋白酶-9与肾脏疾病关系的研究进展

    Institute of Scientific and Technical Information of China (English)

    朱清义; 蒋玉红

    2006-01-01

    基础研究表明,非肿瘤性肾脏疾病的病理改变实质均是细胞外基质(extra cellular matrix,ECM)在肾小球和肾间质中异常堆积的结果。ECM沉积发生机制复杂,其中基质金属蛋白酶(matrix metallo proteinases,MMPs)及其组织抑制因子(tissue inhibitors ofmetalloproteinases,TIMPs)的功能紊乱在ECM转换过程中起关健作用。MMP-9是基质金属蛋白酶家族中的重要成员之一,因其作用底物广泛,表达细胞众多,在国内外研究中备受重视。现将其与肾脏疾病关系的研究进展综述如下。

  16. Advances in Pharmacological Actions and Mechanisms of Doxycycline in the Inhibition of Matrix Metallo-proteinase%多西环素抑制金属蛋白酶药理作用及机制研究进展

    Institute of Scientific and Technical Information of China (English)

    陈薇; 付翔; 黄鹂莺姿; 毛柳珺; 戴莉琳

    2015-01-01

    Doxycycline is a broad-spectrum antimicrobial agent used in clinical for a long time. In recent years, many studies showed that its pharmacological activities of anti-tumor, heart function improvement, nerve protection and antiasthma depended on its inhibition of matrix metalloproteinase ( MMPs) . In the present paper,the new pharmacological actions of doxycycline were reviewed to promote the applications in clinics.%多西环素作为广谱抗菌药物长期应用于临床,近年来很多研究表明多西环素通过抑制金属蛋白酶( MMPs)而具有抗肿瘤、改善心功能,保护神经,平喘等多种药理活性。本文对多西环素新的药理作用进行综述,从而促进其在临床的广泛应用。

  17. 166 Assessment of Chronic Spontaneous Urticaria by Serum-Induced TNF & ALPHA; and MMP-9 Release

    OpenAIRE

    Falkencronec, Sidsel; Poulsen, Lars; Maurer, Marcus; Bindslev-Jensen, Carsten; Skov, Per Stahl

    2012-01-01

    Background Previous studies from our group have demonstrated that IgE-mediated basophil activation leads to release of TNFα that in turn can induce matrix metallo-proteinase-9 (MMP-9) release from monocytes. We wished to investigate if serum from chronic spontaneous urticaria-patients with auto-antibodies against IgE/IgE-receptor could induce TNFα and MMP-9 release from donor PBMCs, and if release levels could be used to assess severity and activity of chronic spontaneous urticaria (CSU). Met...

  18. Matrix metalloproteinase-12 (MMP-12) in osteoclasts

    DEFF Research Database (Denmark)

    Hou, Peng; Troen, Tine; Ovejero, Maria C

    2004-01-01

    Osteoclasts require matrix metalloproteinase (MMP) activity and cathepsin K to resorb bone, but the critical MMP has not been identified. Osteoclasts express MMP-9 and MMP-14, which do not appear limiting for resorption, and the expression of additional MMPs is not clear. MMP-12, also called...

  19. Matrix metalloproteinase-12 (MMP-12) in osteoclasts

    DEFF Research Database (Denmark)

    Hou, Peng; Troen, Tine; Ovejero, Maria C

    2004-01-01

    and resorption. Furthermore, we investigated the role of MMP-12 in bone resorption and osteoclast recruitment by comparing MMP-12 knockout and wild-type mice in specialized culture models known to depend on MMP activity, as well as in the ovariectomy model, and we did not find any indication for a limiting role......Osteoclasts require matrix metalloproteinase (MMP) activity and cathepsin K to resorb bone, but the critical MMP has not been identified. Osteoclasts express MMP-9 and MMP-14, which do not appear limiting for resorption, and the expression of additional MMPs is not clear. MMP-12, also called....... Northern blots show that highly purified rabbit osteoclasts in culture express MMP-12 at the same level as macrophages, whereas in situ hybridizations performed on rabbit bone do not show any MMP-12 expression in osteoclasts whatever the bone type. In contrast, in situ hybridizations performed on mouse...

  20. MMP-2、MMP-3、MMP-9和 TIMP-1评价膝关节骨性关节炎的临床研究%The value of MMP-2,MMP-3,MMP-9 and TIMP-1 levels in the evaluation of knee joint osteoarthritis

    Institute of Scientific and Technical Information of China (English)

    贺占坤; 沈杰威

    2013-01-01

    目的:研究膝关节骨性关节炎(OA)患者关节液中基质金属蛋白酶(MMP)-2、MMP-3、MMP-9和基质金属蛋白酶抑制剂-1(T IM P-1)4种蛋白水平,并探讨其与关节损伤程度及患者预后的关系。方法52例患者均给予关节镜下清理术联合玻璃酸钠、双醋瑞因的治疗方法,于治疗前后采用 ELISA 检测52例膝关节 OA 患者及10例症状轻、X线等影像阴性者关节液中MMP-2、MMP-3、MMP-9和TIMP-1的含量并进行相关分析,并在关节镜下对膝关节软骨损伤程度进行评价。结果膝关节OA患者关节液中MMP-2、MMP-3、MMP-9和TIMP-1的含量均明显高于对照组(P<0.01)。关节液中MMP-2、MMP-3、MMP-9和TIMP-1的含量与关节软骨损伤程度呈正相关,随着病情的好转,关节液中MMP-2、MMP-3、MMP-9和TIMP-1的含量也随之降低。结论检测关节液中MMP-2、MMP-3、MMP-9和TIMP-1水平对膝关节OA的早期诊断、病情程度判断、预后评价有一定的意义。%Objective To explore the relationship between the level of matrix metallo-proteinase(MMP)-2 ,MMP-3 ,MMP-9 and matrix metallo-proteinase inhibitor-1(TIMP-1) in the synovial fluid of the patients with knee joint osteoarthritis (OA) and the de-gree of articular cartilage injury and prognosis .Methods 52 patients(knee OA group) were given arthroscopic debridement com-bined with sodium hyaluronate ,diacerein .The levels of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in synovial fluid were detected in 52 patients with knee OA and 10 normal controls(control group) by enzyme-linked immunosorbent assay .The degree of cartilage inju-ry was assessed with arthroscopy .Results The level of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in knee OA group were significantly higher than those of the control group(all P<0 .01) .The levels of MMP-2 ,MMP-3 ,MMP-9 and TIMP-1 in the synovial fluid were positively correlated with the degree of articular cartilage injury ,and the levels of MMP-2 ,MMP-3 ,MMP-9

  1. Assessment of chronic spontaneous urticaria by serum-induced tumor necrosis factor alpha and matrix metalloproteinase-9 release

    DEFF Research Database (Denmark)

    Falkencrone, Sidsel; Bindslev-Jensen, Carsten; Skov, Per Stahl

    BACKGROUND Previous studies from our group have demonstrated that IgE-mediated basophil activation leads to release of TNFα that in turn can induce matrix metallo-proteinase-9 (MMP-9) release from monocytes. We wished to investigate if serum from chronic spontaneous urticaria-patients with auto......-antibodies against IgE/IgE-receptor could induce TNFα and MMP-9 release from donor PBMCs, and if release levels could be used to assess severity and activity of chronic spontaneous urticaria (CSU). METHODS Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood from healthy donors and basophils...... observed to induce highly significant MMP-9 and TNFα release from donor PBMCs when compared to sera from healthy controls (pUrticaria assessment score (UAS) did not appear to correlate with release levels for histamine, TNFa or MMP-9 in either group but in the ASST+ group, the ASST score appeared...

  2. Dimerization of matrix metalloproteinase-2 (MMP-2): functional implication in MMP-2 activation.

    Science.gov (United States)

    Koo, Bon-Hun; Kim, Yeon Hyang; Han, Jung Ho; Kim, Doo-Sik

    2012-06-29

    Matrix metalloproteinase-2 (MMP-2) functions in diverse biological processes through the degradation of extracellular and non-extracellular matrix molecules. Because of its potential for tissue damage, there are several ways to regulate MMP-2 activity, including gene expression, compartmentalization, zymogen activation, and enzyme inactivation by extracellular inhibitors. Enzyme regulation through zymogen activation is important for the regulation of MMP-2 activity. In our previous studies, we showed that thrombin directly cleaved the propeptide of MMP-2 at specific sites for enzyme activation. We also demonstrated that heparan sulfate was required for thrombin-mediated activation of pro-MMP-2 by binding to thrombin, presumably through conformational changes at the active site of the enzyme. This suggests a regulatory mechanism for thrombin-mediated activation of pro-MMP-2. In this study, we found that MMP-2 formed a reduction-sensitive homodimer in a controlled manner and that Ca(2+) ion was essential for homodimerization of MMP-2. Homodimerization was not associated with protein kinase C-mediated phosphorylation of MMP-2. MMP-2 formed a homodimer through an intermolecular disulfide bond between Cys(102) and the neighboring Cys(102). Homodimerization of MMP-2 enhanced thrombin-mediated activation of pro-MMP-2. Moreover, the MMP-2 homodimer could cleave a small peptide substrate without removal of the propeptide. Taken together, our experimental data suggest a novel regulatory mechanism for pro-MMP-2 activation that is modulated through homodimerization of MMP-2.

  3. Amsacrine suppresses matrix metalloproteinase-2 (MMP-2)/MMP-9 expression in human leukemia cells.

    Science.gov (United States)

    Liu, Wen-Hsin; Chen, Ying-Jung; Chien, Jen-Hung; Chang, Long-Sen

    2014-05-01

    This study explores the suppression mechanism of amsacrine (4-(9-Acridinylamino)-N-(methanesulfonyl)-m-anisidine hydrochloride) on matrix metalloproteinase-2 (MMP-2) and MMP-9 expression in human leukemia cells. Amsacrine attenuated cell invasion with decreased MMP-2/MMP-9 protein expression and mRNA levels in U937, Jurkat, HL-60, K562, KU812, and MEG-01 cells. Moreover, amsacrine reduced both MMP-2/MMP-9 promoter luciferase activity and MMP-2/MMP-9 mRNA stability in leukemia cells. Studies on amsacrine-treated U937 cells revealed that amsacrine-elicited ROS generation induced JNK and p38 MAPK activation but reduced the phospho-ERK level. Amsacrine-induced ERK inactivation and p38 MAPK/JNK activation were demonstrated to suppress MMP-2/MMP-9 promoter luciferase activity and promote MMP-2/MMP-9 mRNA decay, respectively. p38 MAPK/JNK activation led to up-regulation of protein phosphatase 2A catalytic subunit α (PP2Acα) in amsacrine-treated U937 cells. Okadaic acid (PP2A inhibitor) treatment increased MMP-2/MMP-9 mRNA stability in amsacrine-treated cells, whereas PP2Acα over-expression increased MMP-2/MMP-9 mRNA decay. Amsacrine-induced MMP-2/MMP-9 down-regulation was also related to PP2Acα up-regulation on Jurkat, HL-60, K562, KU812, and MEG-01 cells. Collectively, our data indicate that amsacrine induces MMP-2/MMP-9 down-regulation via simultaneous suppression of genetic transcription and mRNA stability in human leukemia cells.

  4. Activation of matrix metalloproteinase-2 (MMP-2) by membrane type 1 matrix metalloproteinase through an artificial receptor for proMMP-2 generates active MMP-2.

    Science.gov (United States)

    Nishida, Yuki; Miyamori, Hisashi; Thompson, Erik W; Takino, Takahisa; Endo, Yoshio; Sato, Hiroshi

    2008-11-01

    The suggested model for pro-matrix metalloproteinase-2 (proMMP-2) activation by membrane type 1 MMP (MT1-MMP) implicates the complex between MT1-MMP and tissue inhibitor of MMP-2 (TIMP-2) as a receptor for proMMP-2. To dissect this model and assess the pathologic significance of MMP-2 activation, an artificial receptor for proMMP-2 was created by replacing the signal sequence of TIMP-2 with cytoplasmic/transmembrane domain of type II transmembrane mosaic serine protease (MSP-T2). Unlike TIMP-2, MSP-T2 served as a receptor for proMMP-2 without inhibiting MT1-MMP, and generated TIMP-2-free active MMP-2 even at a low level of MT1-MMP. Thus, MSP-T2 did not affect direct cleavage of the substrate testican-1 by MT1-MMP, whereas TIMP-2 inhibited it even at the level that stimulates proMMP-2 processing. Expression of MSP-T2 in HT1080 cells enhanced MMP-2 activation by endogenous MT1-MMP and caused intensive hydrolysis of collagen gel. Expression of MSP-T2 in U87 glioma cells, which express a trace level of endogenous MT1-MMP, induced MMP-2 activation and enhanced cell-associated protease activity, activation of extracellular signal-regulated kinase, and metastatic ability into chick embryonic liver and lung. MT1-MMP can exert both maximum MMP-2 activation and direct cleavage of substrates with MSP-T2, which cannot be achieved with TIMP-2. These results suggest that MMP-2 activation by MT1-MMP potentially amplifies protease activity, and combination with direct cleavage of substrate causes effective tissue degradation and enhances tumor invasion and metastasis, which highlights the complex role of TIMP-2. MSP-T2 is a unique tool to analyze physiologic and pathologic roles of MMP-2 and MT1-MMP in comparison with TIMP-2.

  5. Metallo-proteinase from the seedlings of kale (Brassica oleracea L. var. sabellica): : Preparation, partial characterization and substrate specificity.

    Science.gov (United States)

    Wilimowska-Pelc, A; Dryjański, M; Zal, T; Wilusz, T

    1991-10-01

    Metallo-proteinase from 8-d-old seedlings of kale was isolated. The enzyme was extracted with 1% NaCl, concentrated by ammonium sulfate and finally purified by high-performance liquid chromatography. The isolated enzyme had a molecular weight of 22.4 kDa and showed a maximum activity at pH 9.0 using casein as a substrate. Proteolytic activity of proteinase was inhibited by chelators. The inhibition by ethylenediaminetetraacetate (EDTA) was abolished by some divalent metals ions, especially by Zn(2+). The enzyme showed activity against the synthetic peptides Suc-Ala-Ala-Pro-Leu-pNA and Suc-Ala-Ala-Pro-Phe-pNA, and hydrolized the following peptide bonds in the oxidized insulin B-chain: Leu6-Cya7, Leu15-Tyr16, Leu17-Val18 and Phe25-Tyr26.

  6. Expressions of MMP-2 and TIMP-2 in lipopolysaccharide-induced acute lung injury of neonatal rats%MMP-2和TIMP-2在脂多糖诱导的新生大鼠急性肺损伤中表达的变化

    Institute of Scientific and Technical Information of China (English)

    殷静; 杨莉

    2009-01-01

    目的 探讨基质金属蛋白酶-2(matrix metallo proteinase-2,MMP-2)及其组织抑制剂(tissue inhibitors of metallo proteinase-2,TIMP-2)在脂多糖(LPS)诱导的新生大鼠急性肺损伤中的作用.方法 选择30只新生7日龄SD大鼠,随机分为生理盐水对照组(n=6)和急性肺损伤组(n=24),后者进一步分为1、2、4及6h亚组,每组6只.以LPS 腹腔注射建立急性肺损伤模型,观察注射LPS后不同时间点大鼠肺组织的病理改变,用免疫组织化学法和逆转录-聚合酶链反应(RT-PCR)法检测大鼠肺组织中MMP-2和TIMP-2的蛋白及mRNA表达.结果 病理学检查证实新生大鼠急性肺损伤模型复制成功.与生理盐水对照组比较,急性肺损伤组注射LPS后MMP-2蛋白及mRNA均表达增加,4~6h达高峰,差异有统计学意义(均P0.05).结论 LPS致新生大鼠的急性肺损伤存在MMP-2和TIMP-2的表达失衡,进而可能通过破坏基底膜参与急性肺损伤的病理过程.

  7. Expression and inhibition of matrix metalloproteinase (MMP)-8, MMP-9 and MMP-12 in early colonic anastomotic repair

    DEFF Research Database (Denmark)

    Krarup, Peter-Martin; Eld, Mikkel; Heinemeier, Katja Maria;

    2013-01-01

    Submucosal collagen is paramount for colonic anastomotic integrity. Matrix metalloproteinases (MMPs) mediate collagen degradation that increases the risk of wound dehiscence. Although broad-spectrum MMP inhibitors are beneficial for anastomotic strength, they can cause adverse reactions. Knowledg...

  8. Matrix metalloproteinase-2 (MMP-2) and MMP-9 expression in invasive ductal carcinoma of the breast.

    Science.gov (United States)

    Sullu, Yurdanur; Demirag, Guzin G; Yildirim, Arzu; Karagoz, Filiz; Kandemir, Bedri

    2011-12-15

    Matrix metalloproteinase-2 (MMP-2) and MMP-9 are gelatinases that play a role in the invasion and metastasis of cancer through the destruction of the basal membrane and extracellular matrix. In this study, we investigated the immunohistochemical expression of MMP-2 and MMP-9 and the correlation between the expression levels and prognostic clinicopathological parameters in 140 patients with invasive ductal carcinoma (IDC). The staining scores for MMP-9 were negative in 21 cases (15%), mild in 27 cases (19%), and strong in 92 cases (66%). MMP-9 expression was increased in high-grade (p=0.001), triple-negative (ER, PR, HER2 negative) (p=0.006), and ER-negative tumors (p=0.004) and tumors with distant metastases (p=0.028). MMP-9 expression was increased in cases with HER2 over-expression/amplification, but no statistically significant difference was found (p=0.215). No correlation was found between lymph node metastasis or tumor size and MMP-9 expression (p=0.492 and p=0.448, respectively). The staining scores for MMP-2 in 140 cases were negative in 10 cases (7%), mild in 25 cases (18%), and strong in 105 cases (75%). MMP-2 expression was increased in ER-negative and high-grade tumors in the lymph node-negative group (p=0.025 and 0.026, respectively). High MMP-9 expression was associated with a shorter disease-free survival and overall survival times (p=0.042 and p=0.046, respectively). In conclusion, increased MMP-9 expression is related to poor prognostic clinicopathological factors in IDC, and hence, it can be utilized as a supplementary prognostic marker. The role of MMP-2 expression in the prognosis of IDC is rather limited.

  9. 哈萨克族食管癌中MMP-1、MMP-2、MMP-7、TIMP-1和MTA1的表达及其临床病理意义%The expression and significance of MMP.1, MMP.2, MMP.7,TIMP-1 and MTA1 in esophageal cancer of Kazaks

    Institute of Scientific and Technical Information of China (English)

    李秀梅; 陈艳; 王洪江; 庞作良; 李卉; 姜孝芳; 古丽努尔·木哈依; 谌宏鸣; 李惠武

    2011-01-01

    目的:探讨在新疆哈萨克族食管癌组织中基质金属蛋白酶1、2、7 (MMP-1、MMP-2、MMP-7)、基质金属蛋白酶抑制因子1(TIMP-1)和肿瘤转移相关基因1(MTA1)mRNA的表达及其临床意义.方法:采用RT-PCR方法检测75例哈萨克族食管癌标本中MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA的表达水平,分析其表达与临床病理特征的关系.结果:MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA在哈萨克族食管癌组织中的表达较正常组织增高,差异均具有统计学意义(P均<0.05);且MMP-2、MMP-7、MTA1 mRNA的表达与淋巴结转移有关(P<0.05),MMP-1、MMP-7 mRNA的表达与临床分期有关(P<0.05);TIMP-1与MMP-1、MMP-7、MTA1的表达呈正相关(r=0.446、0.458、0.333,P均<0.01).结论:MMP-1、MMP-2、MMP-7、TIMP-1和MTA1 mRNA表达上调共同参与了哈萨克族食管癌的发生发展过程;MMP-2、MMP-7和MTA1可能是哈萨克族食管癌发生侵袭、转移的主导因素.%OBJECTIVE:To study expressions of matrix metalloproteinasel, 2, 7 ( MMP-1, MMP-2, MMP-7), tissue inhibitor of matrix metallo proteinase 1(TIMP-1) and metastasis associated gene 1 (MTA1)in esophageal cancer tissue and normal tissue among Kazaks and to investigate the relationship between their expressions and clinical pathological features. METHODS: Using RT-PCR to detect the expressions of MMP-1, MMP-2, MMP-7, TIMP-1 and MTA1 in 75 esophageal cancer specimens. RESULTS: The mRNA expressions of MMP-1, MMP-2, MMP-7 , TIMP-1 and MTA1 in cancer tissue were higher than those of normal tissues(P<0.05). While the expression levels of MMP-2, MMP-7 and MTA1 mRNA were significantly related to the lymph node metastasis. The expression levels of MMP-1, MMP-7 mRNA were significantly related to the TNM stages(P < 0.05). The expression of TIMP-1 in the Kazak's esophageal cancer was positively related to the expressions of MMP-1, MMP-7, MTA1 (r=0.446、 0.458、 0.333, respectively, P<0.01). CONCLUSION: The overexpressions of MMP

  10. Melanoma cell therapy: Endothelial progenitor cells as shuttle of the MMP12 uPAR-degrading enzyme

    Science.gov (United States)

    Laurenzana, Anna; Biagioni, Alessio; D'Alessio, Silvia; Bianchini, Francesca; Chillà, Anastasia; Margheri, Francesca; Luciani, Cristina; Mazzanti, Benedetta; Pimpinelli, Nicola; Torre, Eugenio; Danese, Silvio; Calorini, Lido; Rosso, Mario Del; Fibbi, Gabriella

    2014-01-01

    The receptor for the urokinase-type plasminogen activator (uPAR) accounts for many features of cancer progression, and is therefore considered a target for anti-tumoral therapy. Only full length uPAR mediates tumor progression. Matrix-metallo-proteinase-12 (MMP12)-dependent uPAR cleavage results into the loss of invasion properties and angiogenesis. MMP12 can be employed in the field of “targeted therapies” as a biological drug to be delivered directly in patient's tumor mass. Endothelial Progenitor Cells (EPCs) are selectively recruited within the tumor and could be used as cellular vehicles for delivering anti-cancer molecules. The aim of our study is to inhibit cancer progression by engeneering ECFCs, a subset of EPC, with a lentivirus encoding the anti-tumor uPAR-degrading enzyme MMP12. Ex vivo manipulated ECFCs lost the capacity to perform capillary morphogenesis and acquired the anti-tumor and anti-angiogenetic activity. In vivo MMP12-engineered ECFCs cleaved uPAR within the tumor mass and strongly inhibited tumor growth, tumor angiogenesis and development of lung metastasis. The possibility to exploit tumor homing and activity of autologous MMP12-engineered ECFCs represents a novel way to combat melanoma by a “personalized therapy”, without rejection risk. The i.v. injection of radiolabelled MMP12-ECFCs can thus provide a new theranostic approach to control melanoma progression and metastasis. PMID:25003596

  11. Melanoma cell therapy: Endothelial progenitor cells as shuttle of the MMP12 uPAR-degrading enzyme.

    Science.gov (United States)

    Laurenzana, Anna; Biagioni, Alessio; D'Alessio, Silvia; Bianchini, Francesca; Chillà, Anastasia; Margheri, Francesca; Luciani, Cristina; Mazzanti, Benedetta; Pimpinelli, Nicola; Torre, Eugenio; Danese, Silvio; Calorini, Lido; Del Rosso, Mario; Fibbi, Gabriella

    2014-06-15

    The receptor for the urokinase-type plasminogen activator (uPAR) accounts for many features of cancer progression, and is therefore considered a target for anti-tumoral therapy. Only full length uPAR mediates tumor progression. Matrix-metallo-proteinase-12 (MMP12)-dependent uPAR cleavage results into the loss of invasion properties and angiogenesis. MMP12 can be employed in the field of "targeted therapies" as a biological drug to be delivered directly in patient's tumor mass. Endothelial Progenitor Cells (EPCs) are selectively recruited within the tumor and could be used as cellular vehicles for delivering anti-cancer molecules. The aim of our study is to inhibit cancer progression by engeneering ECFCs, a subset of EPC, with a lentivirus encoding the anti-tumor uPAR-degrading enzyme MMP12. Ex vivo manipulated ECFCs lost the capacity to perform capillary morphogenesis and acquired the anti-tumor and anti-angiogenetic activity. In vivo MMP12-engineered ECFCs cleaved uPAR within the tumor mass and strongly inhibited tumor growth, tumor angiogenesis and development of lung metastasis. The possibility to exploit tumor homing and activity of autologous MMP12-engineered ECFCs represents a novel way to combat melanoma by a "personalized therapy", without rejection risk. The i.v. injection of radiolabelled MMP12-ECFCs can thus provide a new theranostic approach to control melanoma progression and metastasis.

  12. MMP-8及TIMP-1在慢性牙周炎患者龈沟液中的表达和意义%Expression of MMP-8 and TIMP-1 in gingival crevicular fluid of chronic periodontitis patients

    Institute of Scientific and Technical Information of China (English)

    王丽琴; 冯坤; 孙利

    2014-01-01

    目的 比较慢性牙周炎患者和健康人群龈沟液中基质金属蛋白酶-8(matrix metallo proteinase-8,MMP-8)和其抑制剂基质金属蛋白酶组织抑制剂-1(tissue inhibitor of matrix metalloproteinase-1,TIMP-1)的水平.方法 临床采集30例慢性牙周炎患者和30例正常人群的龈沟液,采用双抗体夹心ABC-ELISA法检MMP-8、TIMP-1含量.结果 慢性牙周炎组MMP-8水平高于正常对照组,差异有统计学意义.TIMP-1水平两组间差异无统计学意义.结论 慢性牙周炎可提高龈沟液MMP-8水平,而MMP-8可能加重牙周组织的损伤.

  13. MMP-2和MMP-9在食管癌中表达的研究%Expressions of MMP-2 and MMP-9 in esophageal cancer

    Institute of Scientific and Technical Information of China (English)

    武志; 潘晓玲; 仲洁

    2013-01-01

    目的 分析基质金属蛋白酶(MMP)-2和MMP-9在食管癌患者血清中的表达与食管癌临床病理因素之间的关系.方法 应用酶联免疫吸附法检测60例食管癌患者血清中MMP-2、MMP-9的含量与患者年龄、性别、组织学类型、临床分期及区域淋巴结转移等临床病理因素之间的关系.结果 对照组血清中MMP-2和MMP-9的浓度均值分别为61.31、87.67 ng/ml,而食管癌患者血清中MMP-2和MMP-9的浓度均值分别121.66、169.73 ng/ml,均高于正常者血清中含量,差异有统计学意义(t=3.015,P=0.007;t=2.037,P=0.04).MMP-2的含量与有无淋巴结转移(t=2.150,P=0.04)及临床分期(t=2.186,P=0.03)有关,MMP-9的含量与有无淋巴结转移(t=2.390,P=0.02)及临床分期(t=2.149,P=0.03)有关.MMP-2和MMP-9的含量均与食管癌患者的年龄、性别、组织学类型无关(均P>0.05).结论 MMP-2和MMP-9可能与食管癌的侵袭转移有关.%Objective To analyze the expression levels of matrix metallo proteinase-2 (MMP-2) and matrix metallo proteinase-9 (MMP-9) in serum of patients with esophageal cancer,and to analyze the interrelations among MMP-2,MMP-9 and clinicopathologic characteristics of esophageal cancer.Methods The levels of serum MMP-2,MMP-9 from 60 patients with esophageal cancer were detected by enzyme linked immunosorbent assay.The interrelations among MMP-2,MMP-9 and clinicopathologic characteristics of esophageal cancer including age,gender,histological type,whether or not lymph nodes metastasis and TNM staging were analyzed.Results The levels of MMP-2 and MMP-9 in control group were 61.31 ng/ml and 87.67 ng/ml,the levels of MMP-2 and MMP-9 in patients with esophageal cancer were 121.66 ng/ml and 169.73 ng/ml.The levels of MMP-2 and MMP-9 were significantly higher in patients with esophageal cancer than that of general population (t =3.015,P =0.007; t =2.037,P =0.04).The level of MMP-2 was significantly associated with whether or not lymph node metastasis (t =2

  14. Expression of gelatinases and tissue inhibitors of metallo- proteinases in the rhesus monkey (Macaca mulatta) corpus luteum

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are believed to play important roles in the formation and regression of corpus luteum (CL). This study is to investigate the expression of gelatinases (MMP-2, -9) and TIMPs in the rhesus monkey CL in both early and late luteal phases and during the early stages of pregnancy. Ovaries were collected from regularly cycling rhesus monkey at D5 and D15 following ovulation and at D12, D18 and D26 of pregnancy. In situ hybridization revealed that in the CL MMP-2 mRNA was expressed during both formation and regression, while MMP-9 mRNA was mainly localized in the late luteal phase. Reduction of MMP-2, -9 transcripts in the CL was observed during pregnancy. MMP-2 mRNA in the CL reduced to an undetectable level at D26 of pregnancy. TIMP-1 mRNA was highly expressed in the CL in both early and late luteal phases and persisted throughout the early stages of pregnancy. Strong signal for TIMP-2 mRNA was also detected in both luteal phases, and the level of TIMP-2 mRNA gradually increased with the progresses of pregnancy. No TIMP-3 mRNA was detected in the macaque CL in this study. In conclusion, these results suggest that MMP-2, -9 and TIMP-1, -2 may have functional roles in rhesus monkey CL. Coordinated expression of MMP-2, -9 and TIMP-2 may play a role in the maintaining of luteal function during early pregnancy. The unchanged expression pattern of TIMP-1 indicates that it may have other functions in the primate CL than inhibition of MMPs.

  15. Angiogenesis in vestibular schwannomas: expression of extracellular matrix factors MMP-2, MMP-9, and TIMP-1

    DEFF Research Database (Denmark)

    Møller, Martin Nue; Werther, Kim; Nalla, Amarnadh;

    2010-01-01

    Vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs) are potent mediators of tumor angiogenesis. It has been demonstrated that vestibular schwannoma VEGF expression correlates with tumor growth pattern, whereas knowledge on the expression of MMPs is lacking. This study t...... targets the angiogenic process by investigation of tumor expression of MMP-2, MMP-9, and tissue inhibitors of metalloproteinase (TIMP)-1. A possible correlation with gender, patient age, symptom duration, tumor size, and the absolute and relative growth rate is explored....

  16. Expression and inhibition of matrix metalloproteinase (MMP)-8, MMP-9 and MMP-12 in early colonic anastomotic repair

    DEFF Research Database (Denmark)

    Krarup, Peter-Martin; Eld, Mikkel; Heinemeier, Katja

    2013-01-01

    Submucosal collagen is paramount for colonic anastomotic integrity. Matrix metalloproteinases (MMPs) mediate collagen degradation that increases the risk of wound dehiscence. Although broad-spectrum MMP inhibitors are beneficial for anastomotic strength, they can cause adverse reactions. Knowledge...... of specific MMPs responsible for the weakening of anastomoses can be used to optimise MMP inhibition therapy. We aimed to quantify transcript and protein levels of multiple MMPs in colonic anastomoses and evaluate the effect of inhibiting the MMPs that displayed the highest expression levels on anastomotic...

  17. Overexpression of matrix metalloproteinase-12 (MMP-12) correlates with radiation-induced lung fibrosis

    Energy Technology Data Exchange (ETDEWEB)

    Jung, Myung Gu; Jeong, Ye Ji; Lee, Haejune [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Lee, Sujae [Hanyang Univ., Seoul (Korea, Republic of)

    2014-05-15

    MMPs are classified into five subgroups: collagenases (MMP-1, MMP-8, MMP-13), gelatinases (MMP-2, MMP-9), stromelysins (MMP-3, MMP-10, MMP-11), as well as metalloelastase (MMP-12), the membrane-type MMPs (MMP14, MMP15), and other MMPS (e. g., MMP-19, and MMP20). MMP-12 (matrix metalloproteinase12), also known as macrophage metalloelastase, was first identified as an elastolytic metalloproteinase secreted by inflammatory macrophages 30 years ago. MMP-12 degrades extracellular matrix (ECM) components to facilitate tissue remodeling. It can degrade elastin and other substrates, such as type IV collagen, fibronectin, laminin, gelatin, vitronectin, entactin, heparin, and chondroitin sulfates. In the lung, MMP-12 is identified in alveolar macrophages of cigarette smokers as an elastolytic MMP. Inactivation of the MMP-12 gene in knockout mice demonstrates a critical role of MMP-12 in smoking-induced chronic obstructive pulmonary disease (COPD). The aim of the present study was to investigate the effects of MMP-12 by radiation in lung, so we evaluate that MMP-12 expression pattern in normal lung tissue and cancer cell following radiation. Radiation induced lung injury most commonly occurs as a result of radiation therapy administered to treat cancer. The present study demonstrates that MMP-12 was highly increased in the lung damaged by radiation Thus, MMP-12 might be of potential relevance as a clinically diagnostic tool and sensitive biomarker for radiation induced lung injury and fibrosis.

  18. Matrix metalloproteinase (MMP)-2 gene polymorphisms affect circulating MMP-2 levels in patients with migraine with aura.

    Science.gov (United States)

    Gonçalves, Flavia M; Martins-Oliveira, Alisson; Lacchini, Riccardo; Belo, Vanessa A; Speciali, Jose G; Dach, Fabíola; Tanus-Santos, Jose E

    2013-01-01

    Matrix metalloproteinases (MMP) are involved in the disruption of blood-brain barrier (BBB) during migraine attacks. In the present study, we hypothesized that two functional polymorphisms (C(-1306)T and C(-735)T) in MMP-2 gene and MMP-2 haplotypes are associated with migraine and modify MMP-2 and tissue inhibitor of MMP (TIMP)-2 levels in migraine. Genotypes for MMP-2 polymorphisms were determined by real time-PCR using Taqman allele discrimination assays. Haplotypes were inferred using the PHASE program. Plasma MMP-2 and TIMP-2 concentrations were measured by gelatin zymography and ELISA, respectively, in 148 healthy women without history of migraine and in 204 women with migraine (153 without aura; MWA, and 51 with aura; MA). Patients with MA had higher plasma MMP-2 concentrations and MMP-2/TIMP-2 ratios than patients with MWA and controls (P0.05), we found that the CC genotype for C(-735)T polymorphism and the CC haplotype were associated with higher plasma MMP-2 concentrations in MA group (P<0.05). Our findings may help to understand the role of MMP-2 and its genetic variants in the pathophysiology of migraine and to identify a particular group of migraine patients with increased MMP-2 levels that would benefit from the use of MMP inhibitors.

  19. Matrix metalloproteinase imbalance in muscle disuse atrophy.

    Science.gov (United States)

    Giannelli, G; De Marzo, A; Marinosci, F; Antonaci, S

    2005-01-01

    Muscle atrophy commonly occurs as a consequence of prolonged muscle inactivity, as observed after cast immobilization, bed rest or space flights. The molecular mechanisms responsible for muscle atrophy are still unknown, but a role has been proposed for altered permeability of the sarcolemma and of the surrounding connective tissue. Matrix metallo-proteinases (MMPs) are a family of enzymes with proteolytic activity toward a number of extracellular matrix (ECM) components; they are inhibited by tissue inhibitors of MMPs (TIMPs). In a rat tail-suspension experimental model, we show that after fourteen days of non-weight bearing there is increased expression of MMP-2 in the atrophic soleus and gastrocnemius and decreased expression of TIMP-2. In the same experimental model the expression of Collagen I and Collagen IV, two main ECM components present in the muscles, was reduced and unevenly distributed in unloaded animals. The difference was more evident in the soleus than in the gastrocnemius muscle. This suggests that muscle disuse induces a proteolytic imbalance, which could be responsible for the breakdown of basal lamina structures such as Collagen I and Collagen IV, and that this leads to an altered permeability with consequent atrophy. In conclusion, an MMP-2/TIMP-2 imbalance could have a role in the mechanism underlying muscle disuse atrophy; more studies are needed to expand our molecular knowledge on this issue and to explore the possibility of targeting the proteolytic imbalance with MMP inhibitors.

  20. Prognostic value of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase N/CD13 in breast cancer patients.

    Science.gov (United States)

    Ranogajec, Irena; Jakić-Razumović, Jasminka; Puzović, Velibor; Gabrilovac, Jelka

    2012-06-01

    The aim of this study was to analyse the expression of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase APN/CD13 in breast carcinoma samples, and their possible prognostic value in breast cancer patients. The expression of MMP-2, MMP-9 and APN/CD13 in tumor cells was analysed in 138 breast carcinomas by immunohistochemical staining of tumor cells using the semiquantitative method for the detection of cytoplasmic and membrane reaction in tumor cells as well as stromal cells positivity. MMP-2 was positive in tumor cells of 52.9% patients and in stromal cells of 74.6% patients, while MMP-9 positive tumor and stromal cells were found in 84.8 and 63.8% patients, respectively. Tumor cell APN/CD13 expression was found in 36.2% patients. Stromal cell MMP-2 expression correlated significantly with tumor size and neoangiogenesis. A positive correlation was also observed between tumor cell MMP-9 expression and hormone receptor status. Stromal cell coexpression of MMP-2/MMP-9 correlated significantly with tumor size. APN/CD13 expression in tumor cells significantly correlated with tumor type and neoangiogenesis. Overall survival was significantly shorter in patients with MMP-2, MMP-2/MMP-9 positive tumor cells, and tended to be shorter in patients with APN/CD13 positive tumor cells. Coexpression of MMP-2/MMP-9 in tumor cells was an independent risk factor for patient survival (OD = 13.9). Our results suggest that MMP-2, MMP-9, APN/CD13 expression and MMP-2/MMP-9 coexpression in combination with other standard prognostic factors can serve as a poor prognostic factor in the evaluation of breast cancer prognosis.

  1. CIL-102 induces matrix metalloproteinase-2 (MMP-2)/MMP-9 down-regulation via simultaneous suppression of genetic transcription and mRNA stability.

    Science.gov (United States)

    Liu, Wen-Hsin; Chen, Yeh-Long; Chang, Long-Sen

    2012-12-01

    This study explores the CIL-102 suppression mechanism on matrix metalloproteinase-2 (MMP-2) and MMP-9 expression in human leukemia K562 cells. CIL-102 attenuated K562 cell invasion with decreased MMP-2/MMP-9 protein expression and mRNA levels. Moreover, CIL-102 reduced luciferase activity of MMP-2/MMP-9 promoter constructs and MMP-2/MMP-9 mRNA stability. CIL-102 treatment induced JNK and p38 MAPK activation but reduced the phospho-ERK level. Transfection of constitutively active MEK1 restored MMP-2 and MMP-9 promoter activity in CIL-102-treated cells, while suppression of p38 MAPK/JNK activation abolished CIL-102-induced MMP-2/MMP-9 mRNA decay. CIL-102-induced p38 MAPK/JNK activation led to protein phosphatase 2A-mediated tristetraprolin (TTP) down-regulation. The reduction in TTP-KH-type splicing regulatory protein (KSRP) complexes formation promoted KSRP-mediated MMP-2/MMP-9 mRNA decay in CIL-102-treated K562 cells. Moreover, CIL-102 reduced invasion and MMP-2/MMP-9 expression in breast and liver cancer cells. Taken together, our data indicate that CIL-102 induces MMP-2/MMP-2 down-regulation via simultaneous suppression of genetic transcription and mRNA stability, and suggest a potential utility for CIL-102 in reducing MMP-2/MMP-9-mediated cancer progression.

  2. The effect of a single nucleotide polymorphism in the matrix metalloproteinase-1 (MMP-1) promoter on force-induced MMP-1 expression in human periodontal ligament cells

    NARCIS (Netherlands)

    Huang, Sheng-Fu; Li, Yu-Hong; Ren, Yi-Jin; Cao, Zheng-Guo; Long, Xing

    2008-01-01

    Matrix metalloproteinase-1 (MMP-1) 1G/2G (-1,607) polymorphisms have been identified and shown to influence the transcription of the MMP-1 gene. In order to compare the expression of MMP-1 with different MMP-1 gene promoter alleles after force loading, human periodontal ligament (PDL) cells were cul

  3. Role of immunohistochemical overexpression of matrix metalloproteinases MMP-2 and MMP-11 in the prognosis of death by ovarian cancer.

    Science.gov (United States)

    Périgny, Martine; Bairati, Isabelle; Harvey, Isabelle; Beauchemin, Michel; Harel, François; Plante, Marie; Têtu, Bernard

    2008-02-01

    Matrix metalloproteinases (MMPs) are enzymes thought to be involved in tumor invasion. We hypothesized that MMP-2 and MMP-11 overexpression was associated with the aggressiveness of ovarian carcinoma. This study was performed on samples from 100 patients with stage III ovarian carcinomas treated surgically between 1990 and 2000. Immunohistochemical staining was performed on ovarian tumors and peritoneal implants using monoclonal antibodies. Overexpression was defined as more than 10% of cells expressing the marker. Multivariate analyses showed that only MMP-2 overexpression by cancer cells in peritoneal implants was associated with a significant risk of death by disease (hazard ratio, 2.65; 95% confidence interval, 1.41-4.97; P =.003). MMP-11 overexpression was not predictive of survival. These results suggest that MMP-2 overexpression by cancer cells in peritoneal implants and not in the primary ovarian cancer is predictive of ovarian cancer prognosis and more likely reflects the presence of particularly aggressive clones of cancer cells.

  4. Matrix metalloproteinase (MMP)-2 and MMP-9 as inflammation markers of Trichinella spiralis and Trichinella pseudospiralis infections in mice.

    Science.gov (United States)

    Bruschi, F; Bianchi, C; Fornaro, M; Naccarato, G; Menicagli, M; Gomez-Morales, M A; Pozio, E; Pinto, B

    2014-10-01

    Trichinella spiralis and Trichinella pseudospiralis exhibit differences in the host-parasite relationship such as the inflammatory response in parasitized muscles. Several studies indicate that matrix metalloproteinases (MMPs) represent a marker of inflammation since they regulate inflammation and immunity. The aim of this study was to evaluate the serum levels of gelatinases (MMP-9 and MMP-2) in mice experimentally infected with T. spiralis or T. pseudospiralis, to elucidate the involvement of these molecules during the inflammatory response to these parasites. Gelatin zymography on SDS polyacrilamide gels was used to assess the serum levels and in situ zymography on muscle histological sections to show the gelatinase-positive cells. In T. spiralis infected mice, the total MMP-9 serum level increased 6 days post-infection whereas, the total MMP-2 serum level increased onward. A similar trend was observed in T. pseudospiralis infected mice but the MMP-9 level was lower than that detected in T. spiralis infected mice. Significant differences were also observed in MMP-2 levels between the two experimental groups. The number of gelatinase positive cells was higher in T. spiralis than in T. pseudospiralis infected muscles. We conclude that MMP-9 and MMP-2 are markers of the inflammatory response for both T. spiralis and T. pseudospiralis infections.

  5. Expression and regulation of metalloproteinases-2, -9 and tissue inhibitors of metallo- proteinases in rat corpus luteum

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The expression and regulation of metalloproteinases-2, -9 (MMP-2, -9) and their tissue inhibitors TIMP-1, -2, -3 mRNA were studied in this experiment. In the PMSG- hCG primed pseudopregnant rat, MMP-2, -9 mRNA levels were the highest at Day 1, decreased from Day 4, and reached the minimal level at Day 8, then increased at Day 14; no significant changes were observed in TIMP-2 mRNA expression from Day 1 to Day 14; TIMP-3 mRNA expression was the lowest at Day 1, increased from Day 4, reached the maximal level at Day 8, and persisted to Day 14. TNF-αcould significantly increase the expression of MMP-2, -9 and TIMP-1 mRNA in the in vitro perfused pseudopregnant CL, and decrease the expression of TIMP-3 mRNA, but had no effect on TIMP-2 mRNA expression. The results indicate that MMP-2, -9 and TIMP-1, -2, -3 might be involved in the regulation of CL function and maintenance of CL structure via their coordinated gene expression. TNF-α could inhibit luteal regression via increasing MMP-2, -9 and TIMP-1 mRNA in the in vitro perfused pseudopregnant ovary.

  6. Matrix metalloproteinases 2 and 9 and MMP9/NGAL complex activity in women with PCOS.

    Science.gov (United States)

    Ranjbaran, Javad; Farimani, Marzieh; Tavilani, Heidar; Ghorbani, Marzieh; Karimi, Jamshid; Poormonsefi, Faranak; Khodadadi, Iraj

    2016-04-01

    It is believed that matrix metalloproteinases (MMPs) play important roles in follicular development and pathogenesis of polycystic ovary syndrome (PCOS). However, conflicting results are available about the alteration of MMP2 and MMP9 concentrations or activities in PCOS. In fact, there is no study entirely investigating both concentration and activity of these MMPs and serum levels of their tissue inhibitors TIMP2 and TIMP1, as well as lipocalin-bound form of MMP9 (MMP9/NGAL). Therefore, the thoroughness of previous studies is questionable. This study was conducted to determine circulatory concentration of MMP2, MMP9, MMP9/NGAL complex, TIMP1 and TIMP2 as well as gelatinase activities of MMP2, MMP9 and MMP9/NGAL complex in women with PCOS and controls. Mean age and BMI as well as serum levels of total cholesterol, triacylglycerol, HDL-C, LDL-C, fasting blood sugar (FBS), insulin, estradiol and sex hormone-binding globulin did not differ between groups, whereas a marked decrease in FSH and significant increases in LH, LH/FSH ratio, testosterone and free androgen index were observed. Women with PCOS and controls showed closed concentrations of MMP2, MMP9, MMP9/NGAL, TIMP1 and TIMP2. Gelatinase activity of MMP9 was found significantly higher in PCOS than in controls (64.53±15.32 vs 44.61±18.95 respectively) while patients and healthy subjects showed similar activities of MMP2 and MMP9/NGAL complex. Additionally, PCOS patients showed a higher MMP9/TIMP1 ratio compared with control women. Direct correlations were also observed between circulatory MMP9 level and the concentration and activity of MMP9/NGAL complex. In conclusion, based on the results of present study, we believe that MMP9 may be involved in the pathogenesis of PCOS.

  7. Expression and prognostic impact of matrix metalloproteinase-2 (MMP-2) in astrocytomas

    DEFF Research Database (Denmark)

    Ramachandran, Rahimsan K.; Sørensen, Mia D.; Aaberg-Jessen, Charlotte

    2017-01-01

    of this tumor. Matrix metalloproteinase-2 (MMP-2) is an extracellular matrix degrading enzyme which has been shown to play important roles in different cancers. The aim of this study was to investigate the expression and prognostic potential of MMP-2 in astrocytomas. Tissue samples from 89 patients diagnosed.......033). We found a positive correlation between MMP-2 and tissue inhibitor of metalloproteinases-1 (TIMP-1), and combined MMP-2 and TIMP-1 had stronger prognostic value than MMP-2 alone also when adjusting for age and gender (HR 2.78; 95% CI 1.30-5.92; p = 0.008). These findings were validated...

  8. Expression of matrix metalloproteinases (MMP-2 、MMP-9) in the pancreatic carcinoma%胰腺癌组织中MMP-2、MMP-9的表达

    Institute of Scientific and Technical Information of China (English)

    吴俊本; 张洁; 成丕光; 王树静; 巩本刚; 徐怀勇

    2012-01-01

    目的 检测胰腺癌组织基质金属蛋白酶( MMP-2,MMP-9)的表达,分析其与临床病理特征的关系.方法 应用免疫组化SP法检测30例胰腺癌及配对癌旁胰腺组织以及11例慢性胰腺炎、6例正常胰腺组织中MMP-2、MMP-9的表达,运用统计软件SPSS 12.0分析其与临床病理特征的相关性.结果 正常胰腺组织均无MMP-2及MMP-9的表达.慢性胰腺炎组织MMP-2、MMP-9的阳性表达率均为18.2% (2/11).胰腺癌组织MMP-2、MMP-9的阳性表达率分别为63.3% (19/30)和56.7% (17/30);配对癌旁组织的阳性表达率分别为23.3% (7/30)和40.0%( 12/30).胰腺癌组织的MMP-2及MMP-9阳性表达率均显著高于配对癌旁组织(P<0.05);胰腺癌组织及癌旁组织中MMP-2及MMP-9的表达均显著高于慢性胰腺炎组织和正常胰腺组织(P值均<0.05).MMP-2、MMP-9的表达与胰腺癌临床分期、肿瘤大小、分化程度及淋巴结转移相关.结论 胰腺癌组织中MMP-2、MMP-9呈高表达,其高表达与肿瘤的恶性程度相关.%Objective To study the expression of matrix metallopro-teinase-2 (MMP-2),matrix metalloproteinase-9 (MMP-9) in the pancreatic carcinomas. Methods MMP-2,MMP-9 expression were detected by immunohistochemistry in surgically resected specimens ( cancer tissues,cancer-adjacent tissues and normal tissues) from 30 PC patients,11 pancreatitis patients and 6 normal patients.the results were analyzed combined with clinical pathologic characteristics.The data was analyzed by Chi-Square test.Results There was no expression of MMP-2,MMP-9 in normal pancreatic tissues.Both of MMP-2 and MMP-9 expressions were 18.2% in chronic pancreatitis titssues.Expression of MMP-2,MMP-9 were higher in cancer tissues than in cancer-adjacent tissues(63.3% vs 23.3%,56.7% vs 40.0%,P <0.05).There were positive correlation between MMP-2,MMP-9 expression and lymph nodal metastasis,tumor sizes,clinical stage and differentiation of tumor(P <0.05).Conclusions

  9. Targeting a single function of the multifunctional matrix metalloprotease MT1-MMP

    DEFF Research Database (Denmark)

    Ingvarsen, Signe; Porse, Astrid; Erpicum, Charlotte

    2013-01-01

    the enzyme ability to activate proMMP-2 without interfering with the collagenolytic function or the general proteolytic activity of MT1-MMP. Using this antibody, we have shown that the MT1-MMP-catalyzed activation of proMMP-2 is involved in the outgrowth of cultured lymphatic endothelial cells in a collagen......-documented importance in matrix degradation but which takes part in more than one pathway in this regard. In this report, we describe the selective targeting of a single function of this enzyme by means of a specific monoclonal antibody against MT1-MMP, raised in an MT1-MMP knock-out mouse. The antibody blocks...... matrix in vitro, as well as in lymphatic vessel sprouting assayed ex vivo. This is the first example of the complete inactivation of a single function of a multifunctional MMP and the use of this strategy to pursue its role....

  10. Serum levels of matrix metalloproteinases MMP-2 and MMP-9 and their tissue natural inhibitors in breast tumors.

    Science.gov (United States)

    Jinga, D; Stefanescu, Maria; Blidaru, A; Condrea, Ileana; Pistol, Gina; Matache, Cristiana

    2004-01-01

    In this study, the levels of matrix metalloproteinases MMP-2 and MMP-9 were simultaneously analyzed with the levels of their tissue natural inhibitors TIMP-1 and TIMP-2 in sera of patients with breast tumors. At the same time, the activity of these two matrix metalloproteinases was evaluated. The decrease of TIMP-2 level in sera from patients with breast cancer as well as an imbalance between MMP-2 and TIMP-2 in neoplasic processes were found. The serum levels of MMP-2, MMP-9 and TIMP-1 were comparable between the patients with breast cancer and benign tumors. These experimental studied parameters were found to correlate with some of clinicopathological disease variables (TNM or pTNM staging system, tumor size and node invasion) suggesting their potential value for diagnosis and prognosis of breast cancer. Matrix metalloproteinases or their natural inhibitors and tumor markers (CA15.3 and CEA) not correlated between but, each of them correlated with another clinicopathological disease variable, suggesting their usefulness in the evaluation.

  11. MT1-MMP-dependent remodeling of cardiac extracellular matrix structure and function following myocardial infarction.

    Science.gov (United States)

    Koenig, Gerald C; Rowe, R Grant; Day, Sharlene M; Sabeh, Farideh; Atkinson, Jeffrey J; Cooke, Kenneth R; Weiss, Stephen J

    2012-05-01

    The myocardial extracellular matrix (ECM), an interwoven meshwork of proteins, glycoproteins, proteoglycans, and glycosaminoglycans that is dominated by polymeric fibrils of type I collagen, serves as the mechanical scaffold on which myocytes are arrayed for coordinated and synergistic force transduction. Following ischemic injury, cardiac ECM remodeling is initiated via localized proteolysis, the bulk of which has been assigned to matrix metalloproteinase (MMP) family members. Nevertheless, the key effector(s) of myocardial type I collagenolysis both in vitro and in vivo have remained unidentified. In this study, using cardiac explants from mice deficient in each of the major type I collagenolytic MMPs, including MMP-13, MMP-8, MMP-2, MMP-9, or MT1-MMP, we identify the membrane-anchored MMP, MT1-MMP, as the dominant collagenase that is operative within myocardial tissues in vitro. Extending these observations to an in vivo setting, mice heterozygous for an MT1-MMP-null allele display a distinct survival advantage and retain myocardial function relative to wild-type littermates in an experimental model of myocardial infarction, effects associated with preservation of the myocardial type I collagen network as a consequence of the decreased collagenolytic potential of cardiac fibroblasts. This study identifies MT1-MMP as a key MMP responsible for effecting postinfarction cardiac ECM remodeling and cardiac dysfunction.

  12. Captopril and lisinopril only inhibit matrix metalloproteinase-2 (MMP-2) activity at millimolar concentrations.

    Science.gov (United States)

    Kuntze, Luciana B; Antonio, Raquel C; Izidoro-Toledo, Tatiane C; Meschiari, Cesar A; Tanus-Santos, Jose E; Gerlach, Raquel F

    2014-03-01

    Matrix metalloproteinase-2 (MMP-2) shares structural similarities with the angiotensin-converting enzyme (ACE). ACE inhibitors have been described to inhibit MMP-2, but this inhibitory potential was not shown using a highly purified MMP-2. This study aimed to investigate the inhibitory potential of captopril and lisinopril regarding MMP-2 activity. The first objective was to test the potential of captopril to change the pH of the buffer solution. The second objective was to test the direct inhibitory effect of captopril and lisinopril on plasma MMP-2 and on recombinant human MMP-2 (rhMMP-2). The in vitro activity assays included gelatin zymography and a fluorimetric assay. Captopril solubilization significantly decreased the pH of the 50 mM Tris buffer solution at the following concentrations: 2 mM (p MMP-2 and rhMMP-2 showed that inhibition only happened at captopril concentrations ≥ 4 and 1 mM, respectively (p MMP-2 (p MMP-2 are 3 orders of magnitude higher than those present in vivo after drug administration. We also discuss possible pitfalls for gelatinase inhibitory assays (besides the obvious pH problem already cited). In conclusion, this study's data show that captopril and lisinopril did not inhibit MMP-2 directly at the concentrations reached in vivo.

  13. Immunoexpression of matrix metalloproteinase-2 (MMP-2) in epithelial ovarian cancers (EOCs)

    Institute of Scientific and Technical Information of China (English)

    Ibrahim A Abdelazim; Mohannad Lutfi Abu faza; Mohammed Al-Kadi

    2013-01-01

    Objective: To evaluate the relation between matrix metalloproteinase-2 (MMP-2) expression and the clinical and/or pathological parameters of the epithelial ovarian cancers (EOCs). Methods: Forty-two (42) patients with EOCs diagnosed after histopathological examination of the specimens were included in this study. The pathological specimens were additionally stained by immunoperoxidase technique for MMP-2 using a monoclonal antibody against activated MMP 2. The staining intensity of MMP-2 was correlated with the clinical and pathological parameters of the studied cases, including patient's age, surgical stage, histological grade, omental, and lymph node metastasis. Results: The studied cases of EOCs were classified according to the intensity or the degree of MMP-2 expression, as; seven cases (16.7%) negative, eighteen cases (42.8%) weak, seven cases (16.7%) moderate and ten cases (23.8%) intense for MMP-2 staining. There was a significant positive correlation between MMP-2 expression and the histological grades and the surgical stages of the studied EOC (r <1, P<0.05), while, there was no significant relation between MMP-2 expression and the histopathological types of the studied EOCs. MMP-2 expression was significantly high in EOCs with ascites, omental, distant and uterine metastasis, while, there was no significant relation between MMP-2 expressions and lymph node metastasis or bilaterality of the EOCs. Conclusions: MMP-2 expression was associated with advanced, aggressive EOCs and there was direct relation between expression of MMP-2 and degree of invasiveness and metastasis of EOCs.

  14. Matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) in preoperative serum as independent prognostic markers in patients with colorectal cancer.

    Science.gov (United States)

    Dragutinović, Vesna V; Radonjić, Nevena V; Petronijević, Nataša D; Tatić, Svetislav B; Dimitrijević, Ivan B; Radovanović, Nebojša S; Krivokapić, Zoran V

    2011-09-01

    Colorectal cancer is one of the leading causes of cancer related death in developed countries. One of the reasons is the absence of tumor specific diagnostic and prognostic markers. The aim of this study was to examine the correlation of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) expressions in serum and clinicopathological features of the colorectal adenocarcinoma. Another aim was to examine expression of MMP-9 in the tissue of the colorectal carcinoma in MMP-9 serum positive patients. In addition, we tried to establish the correlation between preoperative levels of serum markers (CEA and CA 19-9) and presence of MMP-2 or MMP-9. The study was performed on 32 patients with colorectal adenocarcinoma who underwent surgery and 11 patients in a control group who were operated for benign diseases. The samples were analyzed by SDS-PAGE to determine the molecular mass and SDS-PAGE zymography to determine levels of MMP-2 and MMP-9. Expression of MMP-9 was determined immunohistochemically in the tissue of the colorectal carcinoma of MMP-9 serum positive patients. MMP-2 and MMP-9 levels were increased in the serum of the patients with colorectal cancer compared to the control group. There was significant correlation in MMPs levels among the patients with tumor stage I and II and the patients with tumor stage III and IV. Obtained results did not demonstrate correlation between levels of CEA, CA 19-9 and presence of MMP-2 or MMP-9. MMP-9 expression was positive in 85% of MMP-9 serum positive patients with colorectal carcinoma. The overexpression of MMP-2 and MMP-9 strongly suggests its association with colorectal adenocarcinoma. Detection of MMP-2 and MMP-9 in serum might be useful for identification of patients with higher risk for colorectal cancer recurrence.

  15. Neutrophil Gelatinase-Associated Lipocalin (NGAL), Pro-Matrix Metalloproteinase-9 (pro-MMP-9) and Their Complex Pro-MMP-9/NGAL in Leukaemias

    Energy Technology Data Exchange (ETDEWEB)

    Bouchet, Sandrine; Bauvois, Brigitte, E-mail: brigitte.bauvois@crc.jussieu.fr [INSERM U1138, Université Pierre et Marie Curie, Université Paris-Descartes, Centre de Recherche des Cordeliers, Paris 75006 (France)

    2014-04-04

    Matrix metalloproteinase (MMP)-9 and neutrophil gelatinase-associated lipocalin (NGAL) have gained attention as cancer biomarkers. The inactive zymogen form of MMP-9 (pro-MMP-9) also exists as a disulphide-linked heterodimer bound to NGAL in humans. Leukaemias represent a heterogeneous group of neoplasms, which vary in their clinical behavior and pathophysiology. In this review, we summarize the current literature on the expression profiles of pro-MMP-9 and NGAL as prognostic factors in leukaemias. We also report the expression of the pro-MMP-9/NGAL complex in these diseases. We discuss the roles of (pro)-MMP-9 (active and latent forms) and NGAL in tumour development, and evaluate the mechanisms by which pro-MMP-9/NGAL may influence the actions of (pro)-MMP-9 and NGAL in cancer. Emerging knowledge about the coexpression and the biology of (pro)-MMP-9, NGAL and their complex in cancer including leukaemia may improve treatment outcomes.

  16. Matrix Metalloproteinase-3 (MMP-3) Is an Endogenous Activator of the MMP-9 Secreted by Placental Leukocytes: Implication in Human Labor.

    Science.gov (United States)

    Flores-Pliego, Arturo; Espejel-Nuñez, Aurora; Castillo-Castrejon, Marisol; Meraz-Cruz, Noemi; Beltran-Montoya, Jorge; Zaga-Clavellina, Veronica; Nava-Salazar, Sonia; Sanchez-Martinez, Maribel; Vadillo-Ortega, Felipe; Estrada-Gutierrez, Guadalupe

    2015-01-01

    The activity of matrix degrading enzymes plays a leading role in the rupture of the fetal membranes under normal and pathological human labor, and matrix metalloproteinase-9 (MMP-9) it is considered a biomarker of this event. To gain further insight into local MMP-9 origin and activation, in this study we analyzed the contribution of human placental leukocytes to MMP-9 secretion and explored the local mechanisms of the pro-enzyme activation. Placental blood leukocytes were obtained from women at term gestation without labor and maintained in culture up to 72 h. MMP-9 activity in the culture supernatants was determined by zymography and using a specific substrate. The presence of a potential pro-MMP-9 activator in the culture supernatants was monitored using a recombinant biotin-labeled human pro-MMP-9. To characterize the endogenous pro-MMP-9 activator, MMP-1, -3, -7 and -9 were measured by multiplex assay in the supernatants, and an inhibition assay of MMP-9 activation was performed using an anti-human MMP-3 and a specific MMP-3 inhibitor. Finally, production of MMP-9 and MMP-3 in placental leukocytes obtained from term pregnancies with and without labor was assessed by immunofluorescence. Placental leukocytes spontaneously secreted pro-MMP-9 after 24 h of culture, increasing significantly at 48 h (P≤0.05), when the active form of MMP-9 was detected. Culture supernatants activated the recombinant pro-MMP-9 showing that placental leukocytes secrete the activator. A significant increase in MMP-3 secretion by placental leukocytes was observed since 48 h in culture (P≤0.05) and up to 72 h (P≤0.001), when concentration reached its maximum value. Specific activity of MMP-9 decreased significantly (P≤0.005) when an anti-MMP-3 antibody or a specific MMP-3 inhibitor were added to the culture media. Placental leukocytes from term labor produced more MMP-9 and MMP-3 compared to term non-labor cells. In this work we confirm that placental leukocytes from human term

  17. Matrix Metalloproteinase-3 (MMP-3 Is an Endogenous Activator of the MMP-9 Secreted by Placental Leukocytes: Implication in Human Labor.

    Directory of Open Access Journals (Sweden)

    Arturo Flores-Pliego

    Full Text Available The activity of matrix degrading enzymes plays a leading role in the rupture of the fetal membranes under normal and pathological human labor, and matrix metalloproteinase-9 (MMP-9 it is considered a biomarker of this event. To gain further insight into local MMP-9 origin and activation, in this study we analyzed the contribution of human placental leukocytes to MMP-9 secretion and explored the local mechanisms of the pro-enzyme activation.Placental blood leukocytes were obtained from women at term gestation without labor and maintained in culture up to 72 h. MMP-9 activity in the culture supernatants was determined by zymography and using a specific substrate. The presence of a potential pro-MMP-9 activator in the culture supernatants was monitored using a recombinant biotin-labeled human pro-MMP-9. To characterize the endogenous pro-MMP-9 activator, MMP-1, -3, -7 and -9 were measured by multiplex assay in the supernatants, and an inhibition assay of MMP-9 activation was performed using an anti-human MMP-3 and a specific MMP-3 inhibitor. Finally, production of MMP-9 and MMP-3 in placental leukocytes obtained from term pregnancies with and without labor was assessed by immunofluorescence.Placental leukocytes spontaneously secreted pro-MMP-9 after 24 h of culture, increasing significantly at 48 h (P≤0.05, when the active form of MMP-9 was detected. Culture supernatants activated the recombinant pro-MMP-9 showing that placental leukocytes secrete the activator. A significant increase in MMP-3 secretion by placental leukocytes was observed since 48 h in culture (P≤0.05 and up to 72 h (P≤0.001, when concentration reached its maximum value. Specific activity of MMP-9 decreased significantly (P≤0.005 when an anti-MMP-3 antibody or a specific MMP-3 inhibitor were added to the culture media. Placental leukocytes from term labor produced more MMP-9 and MMP-3 compared to term non-labor cells.In this work we confirm that placental leukocytes from

  18. Neurokinin-1 receptor directly mediates glioma cell migration by up-regulation of matrix metalloproteinase-2 (MMP-2) and membrane type 1-matrix metalloproteinase (MT1-MMP).

    Science.gov (United States)

    Mou, Lingyun; Kang, Yawei; Zhou, Ying; Zeng, Qian; Song, Hongjing; Wang, Rui

    2013-01-04

    Neurokinin-1 receptor (NK1R) occurs naturally on human glioblastomas. Its activation mediates glioma cell proliferation. However, it is unknown whether NK1R is directly involved in tumor cell migration. In this study, we found human hemokinin-1 (hHK-1), via NK1R, dose-dependently promoted the migration of U-251 and U-87 cells. In addition, we showed that hHK-1 enhanced the activity of MMP-2 and the expression of MMP-2 and MT1-matrix metalloproteinase (MMP), which were responsible for cell migration, because neutralizing the MMPs with antibodies decreased cell migration. The involved mechanisms were then investigated. In U-251, hHK-1 induced significant calcium efflux; phospholipase C inhibitor U-73122 reduced the calcium mobilization, the up-regulation of MMP-2 and MT1-MMP, and the cell migration induced by hHK-1, which meant the migration effect of NK1R was mainly mediated through the G(q)-PLC pathway. We further demonstrated that hHK-1 boosted rapid phosphorylation of ERK, JNK, and Akt; inhibition of ERK and Akt effectively reduced MMP-2 induction by hHK-1. Meanwhile, inhibition of ERK, JNK, and Akt reduced the MT1-MMP induction. hHK-1 stimulated significant phosphorylation of p65 and c-JUN in U-251. Reporter gene assays indicated hHK-1 enhanced both AP-1 and NF-κB activity; inhibition of ERK, JNK, and Akt dose-dependently suppressed the NF-κB activity; only the inhibition of ERK significantly suppressed the AP-1 activity. Treatment with specific inhibitors for AP-1 or NF-κB strongly blocked the MMP up-regulation by hHK-1. Taken together, our data suggested NK1R was a potential regulator of human glioma cell migration by the up-regulation of MMP-2 and MT1-MMP.

  19. 前列腺癌中MMP-9和VEGF的表达及相关性研究%An Association Study of MMP-9 and VEGF Protein EXpression in Prostate Cancer

    Institute of Scientific and Technical Information of China (English)

    牛军

    2014-01-01

    目的:探讨基质金属蛋白酶( MMP-9)和血管内皮生长因子( VEGF)在前列腺癌组织中的表达以及两者的关系。方法:应用免疫组织化学技术检测120例前列腺增生( BPH)和120例前列腺癌( PCa)组织中MMP-9和VEG的表达水平,并分析两者表达与前列腺癌临床病理特征的关系。结果:120例PCa中MMP-9和VEGF阳性表达率分别为85.01%、83.32%,与BPH组织进行比较,差异均有统计学意义( P <0.05)。MMP-9和VEGF与PCa的病理分级和临床分期有关,且二者呈正相关。结论:MMP-9和VEGF蛋白在PCa中高表达,两者与前列腺癌的发生、发展相关,可以作为预后的判断指标。%ObjectiVe:To eXplore the eXpression of matriX metallo proteinases -9( MMP -9 )and Vascular endothelial growth factor( VEGF)in prostatic cancer( PCA)and the relations between them. Methods:The eXpression of VEGF and MMP-9 in 120 cases of prostate cancer and 120 case s of BPH were detected by immunohistochemistry,with the relationship of their eXpression with clinical characteristics of prostatic cancer further analyzed. ResuIts:The positiVe rate of MMP-9 and VEGF eX-pression in 120 cases PCA is 85. 01 % and 83. 32 %,respectiVely,significantly different from that in BPH( P <0. 0 5). The eXpression of MMP-9 and VEGF in prostate cancer was positiVely correlated with pathological grading and clinical staging of PCA. ConcIusion:The oVer eXpression of MMP-9 and VEGF in PCA is correlated to the generation and the deVelopment of the prostatic carcinoma,which can be taken as an indeX of the prognosis of PCA.

  20. Ovarian Membrane-Type Matrix Metalloproteinases: Induction of MMP14 and MMP16 During the Periovulatory Period in the Rat, Macaque, and Human1

    Science.gov (United States)

    Puttabyatappa, Muraly; Jacot, Terry A.; Al-Alem, Linah F.; Rosewell, Katherine L.; Duffy, Diane M.; Brännström, Mats; Curry, Thomas E.

    2014-01-01

    ABSTRACT An intrafollicular increase in proteolytic activity drives ovulatory events. Surprisingly, the periovulatory expression profile of the membrane-type matrix metalloproteinases (MT-MMPs), unique proteases anchored to the cell surface, has not been extensively examined. Expression profiles of the MT-MMPs were investigated in ovarian tissue from well-characterized rat and macaque periovulatory models and naturally cycling women across the periovulatory period. Among the six known MT-MMPs, mRNA expression of Mmp14, Mmp16, and Mmp25 was increased after human chorionic gonadotropin (hCG) administration in rats. In human granulosa cells, mRNA expression of MMP14 and MMP16 increased following hCG treatment. In contrast, mRNA levels of MMP16 and MMP25 in human theca cells were unchanged before ovulation but declined by the postovulatory stage. In macaque granulosa cells, hCG increased mRNA for MMP16 but not MMP14. Immunoblotting showed that protein levels of MMP14 and MMP16 in rats increased, similar to their mRNA expression. In macaque granulosa cells, only the active form of the MMP14 protein increased after hCG, unlike its mRNA or the proprotein. By immunohistochemistry, both MMP14 and MMP16 localized to the different ovarian cell types in rats and humans. Treatment with hCG resulted in intense immunoreactivity of MMP14 and MMP16 proteins in the granulosa and theca cells. The present study shows that MMP14 and MMP16 are increased by hCG administration in the ovulating follicle, demonstrating that these MMPs are conserved among rats, macaques, and humans. These findings suggest that MT-MMPs could have an important role in promoting ovulation and remodeling of the ovulated follicle into the corpus luteum. PMID:24920038

  1. Signatures of positive selection at hemopexin (PEX) domain of matrix metalloproteinase-9 (MMP-9) gene

    Indian Academy of Sciences (India)

    Yang Liu; Yang Zhao; Chunlei Lu; Maobin Fu; Tonghai Dou; Xiaoming Tan

    2015-12-01

    Matrix metalloproteinases-9 (MMP-9) is an important cancer-associated, zinc-dependent endopeptidase. To investigate the natural selection hypothesis of MMP-9, the orthologous sequences from 12 vertebrates were compared and a molecular evolution analysis was performed. Results suggest that amino acid residues present in the middle region of the protein are more selectively constrained, whereas amino acid residues in the C-terminal region of the MM~P-9 protein including exon 13 showed lowest conservation level in non-primate species, suggesting that it is an exon with fast evolving rate compared to the others analyzed. InterProScan analysis shows that exon 13 was located in hemopexin (PEX) domain of MM~P-9. Positive selection was detected in PEX domain of MMP-9 protein between human and other species, which indicates that selective pressure may play a role in shaping the function of MM~P-9 in the course of evolution.

  2. Plasma Levels of Matrix Metalloproteinase (MMP)-2, MMP-9 and Tumor Necrosis Factor-α in Chronic Hepatitis C Virus Patients

    Science.gov (United States)

    Abdel-Latif, Mohamed S

    2015-01-01

    Background: In chronic HCV infection, pathological accumulation of the extracellular matrix is the main feature of liver fibrosis; that indicates the imbalanced rate of increased matrix synthesis to decreased breakdown of connective tissue proteins. Matrix metalloproteinases (MMPs) play a crucial role in remodeling of extracellular matrix. It is known that expression of MMPs is regulated by Tumor necrosis factor (TNF)-α. Also, levels of TNF-α in liver and serum are increased in chronic HCV patient. Accordingly, this study aimed to correlate the plasma levels of MMP-2, MMP-9 and TNF-α in chronic HCV patients with the pathogenesis of the liver. Methods: The current study was conducted on 15 fibrotic liver cases with detectable HCV RNA, 10 HCV cirrhotic liver cases, and 15 control subjects of matched age and sex. Plasma MMP-2, MMP-9 and TNF-α were measured by ELISA. Results: Data revealed that the MMP2, MMP9 and TNF-α levels showed a significant elevation in chronic HCV patients compared to control group (p= 0.001). But, no significant correlation was observed in levels of MMP-2, MMP-9, and TNF-α between fibrotic and cirrhotic cases. Conclusions: MMP-2, MMP-9 and TNF-α showed high reproducibility to differentiate chronic HCV patients from control group. On the contrary, MMP-2, MMP-9 and TNF-α were not able to differentiate fibrotic from cirrhotic liver cases. Thus, MMP-2, MMP-9 and TNF-α could not be correlated with the progression of liver disease. Rather they could be used as prognostic markers of liver fibrosis. PMID:26464613

  3. Extra-cellular matrix proteins induce matrix metalloproteinase-1 (MMP-1 activity and increase airway smooth muscle contraction in asthma.

    Directory of Open Access Journals (Sweden)

    Natasha K Rogers

    Full Text Available Airway remodelling describes the histopathological changes leading to fixed airway obstruction in patients with asthma and includes extra-cellular matrix (ECM deposition. Matrix metalloproteinase-1 (MMP-1 is present in remodelled airways but its relationship with ECM proteins and the resulting functional consequences are unknown. We used airway smooth muscle cells (ASM and bronchial biopsies from control donors and patients with asthma to examine the regulation of MMP-1 by ECM in ASM cells and the effect of MMP-1 on ASM contraction. Collagen-I and tenascin-C induced MMP-1 protein expression, which for tenascin-C, was greater in asthma derived ASM cells. Tenascin-C induced MMP-1 expression was dependent on ERK1/2, JNK and p38 MAPK activation and attenuated by function blocking antibodies against the β1 and β3 integrin subunits. Tenascin-C and MMP-1 were not expressed in normal airways but co-localised in the ASM bundles and reticular basement membrane of patients with asthma. Further, ECM from asthma derived ASM cells stimulated MMP-1 expression to a greater degree than ECM from normal ASM. Bradykinin induced contraction of ASM cells seeded in 3D collagen gels was reduced by the MMP inhibitor ilomastat and by siRNA knockdown of MMP-1. In summary, the induction of MMP-1 in ASM cells by tenascin-C occurs in part via integrin mediated MAPK signalling. MMP-1 and tenascin-C are co-localised in the smooth muscle bundles of patients with asthma where this interaction may contribute to enhanced airway contraction. Our findings suggest that ECM changes in airway remodelling via MMP-1 could contribute to an environment promoting greater airway narrowing in response to broncho-constrictor stimuli and worsening asthma symptoms.

  4. Neutrophil-Derived MMP-8 Drives AMPK-Dependent Matrix Destruction in Human Pulmonary Tuberculosis.

    Directory of Open Access Journals (Sweden)

    Catherine W M Ong

    2015-05-01

    Full Text Available Pulmonary cavities, the hallmark of tuberculosis (TB, are characterized by high mycobacterial load and perpetuate the spread of M. tuberculosis. The mechanism of matrix destruction resulting in cavitation is not well defined. Neutrophils are emerging as key mediators of TB immunopathology and their influx are associated with poor outcomes. We investigated neutrophil-dependent mechanisms involved in TB-associated matrix destruction using a cellular model, a cohort of 108 patients, and in separate patient lung biopsies. Neutrophil-derived NF-kB-dependent matrix metalloproteinase-8 (MMP-8 secretion was up-regulated in TB and caused matrix destruction both in vitro and in respiratory samples of TB patients. Collagen destruction induced by TB infection was abolished by doxycycline, a licensed MMP inhibitor. Neutrophil extracellular traps (NETs contain MMP-8 and are increased in samples from TB patients. Neutrophils lined the circumference of human pulmonary TB cavities and sputum MMP-8 concentrations reflected TB radiological and clinical disease severity. AMPK, a central regulator of catabolism, drove neutrophil MMP-8 secretion and neutrophils from AMPK-deficient patients secrete lower MMP-8 concentrations. AMPK-expressing neutrophils are present in human TB lung biopsies with phospho-AMPK detected in nuclei. These data demonstrate that neutrophil-derived MMP-8 has a key role in the immunopathology of TB and is a potential target for host-directed therapy in this infectious disease.

  5. MMP20, KLK4, and MMP20/KLK4 double null mice define roles for matrix proteases during dental enamel formation.

    Science.gov (United States)

    Hu, Yuanyuan; Smith, Charles E; Richardson, Amelia S; Bartlett, John D; Hu, Jan C C; Simmer, James P

    2016-03-01

    Matrix metalloproteinase 20 (MMP20) and kallikrein-related peptidase 4 (KLK4) are secreted proteinases that are essential for proper dental enamel formation. We characterized and compared enamel formed in wild-type, Mmp20 (-/-), Klk4 (-/-), Mmp20 (+/-) Klk4 (+/-), and Mmp20 (-/-) Klk4 (-/-) mice using dissecting and light microscopy, backscattered scanning electron microscopy (bSEM), SEM, microcomputed tomography (μCT), and energy-dispersive X-ray analysis (EDX). Following eruption, fractures were observed on Mmp20 (-/-), Klk4 (-/-), Mmp20 (+/-) Klk4 (+/-), and Mmp20 (-/-) Klk4 (-/-) molars. Failure of the enamel in the Mmp20 (+/-) Klk4 (+/-) molars was unexpected and suggested that digenic effects could contribute to the etiology of amelogenesis imperfecta in humans. Micro-CT analyses of hemimandibles demonstrated significantly reduced high-density enamel volume in the Mmp20 (-/-) and Klk4 (-/-) mice relative to the wild-type, which was further reduced in Mmp20 (-/-) Klk4 (-/-) mice. bSEM images of 7-week Mmp20 (-/-) and Mmp20 (-/-) Klk4 (-/-) mandibular incisors showed rough, pitted enamel surfaces with numerous indentations and protruding nodules. The Mmp20 (+/-) and Mmp20 (+/-) Klk4 (+/-) incisors showed prominent, evenly spaced, horizontal ridges that were more distinct in Mmp20 (+/-) Klk4 (+/-) incisors relative to Mmp20 (+/-) incisors due to the darkening of the valleys between the ridges. In cross sections, the Mmp20 (-/-) and Mmp20 (-/-) Klk4 (-/-) exhibited three distinct layers. The outer layer exhibited a disturbed elemental composition and an irregular enamel surface covered with nodules. The Mmp20 null enamel was apparently unable to withstand the sheer forces associated with eruption and separated from dentin during development. Cells invaded the cracks and interposed between the dentin and enamel layers. MMP20 and KLK4 serve overlapping and complementary functions to harden enamel by removing protein, but MMP20 potentially serves multiple

  6. Matrix metalloproteinase-2 (MMP-2) generates soluble HLA-G1 by cell surface proteolytic shedding.

    Science.gov (United States)

    Rizzo, Roberta; Trentini, Alessandro; Bortolotti, Daria; Manfrinato, Maria C; Rotola, Antonella; Castellazzi, Massimiliano; Melchiorri, Loredana; Di Luca, Dario; Dallocchio, Franco; Fainardi, Enrico; Bellini, Tiziana

    2013-09-01

    Human leukocyte antigen-G (HLA-G) molecules are non-classical HLA class I antigens with an important role in pregnancy immune regulation and inflammation control. Soluble HLA-G proteins can be generated through two mechanisms: alternative splicing and proteolytic release, which is known to be metalloprotease mediated. Among this class of enzymes, matrix metalloproteinases (MMPs) might be involved in the HLA-G1 membrane cleavage. Of particular interest are MMP-2 and MMP-9, which regulate the inflammatory process by cytokine and chemokine modulation. We evaluated the effect of MMP-9 and MMP-2 on HLA-G1 membrane shedding. In particular, we analyzed the in vitro effect of these two gelatinases on the secretion of HLA-G1 via proteolytic cleavage in 221-G1-transfected cell line, in JEG3 cell line, and in peripheral blood mononuclear cells. The results obtained by both cell lines showed the role of MMP-2 in HLA-G1 shedding. On the contrary, MMP-9 was not involved in this process. In addition, we identified three possible highly specific cleavage sites for MMP-2, whereas none were detected for MMP-9. This study suggests an effective link between MMP-2 and HLA-G1 shedding, increasing our knowledge on the regulatory machinery beyond HLA-G regulation in physiological and pathological conditions.

  7. Molecular design of a highly selective and strong protein inhibitor against matrix metalloproteinase-2 (MMP-2).

    Science.gov (United States)

    Higashi, Shouichi; Hirose, Tomokazu; Takeuchi, Tomoka; Miyazaki, Kaoru

    2013-03-29

    Synthetic inhibitors of matrix metalloproteinases (MMPs), designed previously, as well as tissue inhibitors of metalloproteinases (TIMPs) lack enzyme selectivity, which has been a major obstacle for developing inhibitors into safe and effective MMP-targeted drugs. Here we designed a fusion protein named APP-IP-TIMP-2, in which the ten amino acid residue sequence of APP-derived MMP-2 selective inhibitory peptide (APP-IP) is added to the N terminus of TIMP-2. The APP-IP and TIMP-2 regions of the fusion protein are designed to interact with the active site and the hemopexin-like domain of MMP-2, respectively. The reactive site of the TIMP-2 region, which has broad specificity against MMPs, is blocked by the APP-IP adduct. The recombinant APP-IP-TIMP-2 showed strong inhibitory activity toward MMP-2 (Ki(app) = 0.68 pm), whereas its inhibitory activity toward MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, or MT1-MMP was six orders of magnitude or more weaker (IC50 > 1 μm). The fusion protein inhibited the activation of pro-MMP-2 in the concanavalin A-stimulated HT1080 cells, degradation of type IV collagen by the cells, and the migration of stimulated cells. Compared with the decapeptide APP-IP (t½ = 30 min), APP-IP-TIMP-2 (t½ ≫ 96 h) showed a much longer half-life in cultured tumor cells. Therefore, the fusion protein may be a useful tool to evaluate contributions of proteolytic activity of MMP-2 in various pathophysiological processes. It may also be developed as an effective anti-tumor drug with restricted side effects.

  8. Matrix Metalloproteinase-2 (MMP-2) Gene Deletion Enhances MMP-9 Activity, Impairs PARP-1 Degradation, and Exacerbates Hepatic Ischemia and Reperfusion Injury in Mice.

    Science.gov (United States)

    Kato, Hiroyuki; Duarte, Sergio; Liu, Daniel; Busuttil, Ronald W; Coito, Ana J

    2015-01-01

    Hepatic ischemia and reperfusion injury (IRI) is an inflammatory condition and a significant cause of morbidity and mortality after surgery. Matrix metalloproteinases (MMPs) have been widely implicated in the pathogenesis of inflammatory diseases. Among the different MMPs, gelatinases (MMP-2 and MMP-9) are within the most prominent MMPs detected during liver IRI. While the role of MMP-9 in liver damage has been fairly documented, direct evidence of the role for MMP-2 activity in hepatic IRI remains to be established. Due to the lack of suitable inhibitors to target individual MMPs in vivo, gene manipulation is as an essential tool to assess MMP direct contribution to liver injury. Hence, we used MMP-2-/- deficient mice and MMP-2+/+ wild-type littermates to examine the function of MMP-2 activity in hepatic IRI. MMP-2 expression was detected along the sinusoids of wild-type livers before and after surgery and in a small population of leukocytes post-IRI. Compared to MMP-2+/+ mice, MMP-2 null (MMP-2-/-) mice showed exacerbated liver damage at 6, 24, and 48 hours post-reperfusion, which was fatal in some cases. MMP-2 deficiency resulted in upregulation of MMP-9 activity, spontaneous leukocyte infiltration in naïve livers, and amplified MMP-9-dependent transmigration of leukocytes in vitro and after hepatic IRI. Moreover, complete loss of MMP-2 activity impaired the degradation of poly (ADP-ribose) polymerase (PARP-1) in extensively damaged livers post-reperfusion. However, the administration of a PARP-1 inhibitor to MMP-2 null mice restored liver preservation to almost comparable levels of MMP-2+/+ mice post-IRI. Deficient PARP-1 degradation in MMP-2-null sinusoidal endothelial cells correlated with their increased cytotoxicity, evaluated by the measurement of LDH efflux in the medium. In conclusion, our results show for the first time that MMP-2 gene deletion exacerbates liver IRI. Moreover, they offer new insights into the MMP-2 modulation of inflammatory responses

  9. MMP-9 directed shRNAs as relevant inhibitors of matrix metalloproteinase 9 activity and signaling

    Directory of Open Access Journals (Sweden)

    Ewa Nowak

    2013-08-01

    Full Text Available Introduction: The main function of matrix metalloproteinases is the degradation of extracellular matrix components, which is related to changes in the proliferation of cells, their differentiation, motility, and death. MMPs play an important role in physiological processes such as embryogenesis, angiogenesis and tissue remodeling. The increase of MMPs activity is also observed in pathological conditions including tumorigenesis where MMP-2 (gelatinase A and MMP-9 (gelatinase B show the ability to degrade the basement membrane of vessels and they are involved in metastasis. The aim of our study was to verify the changes of MMP-9 enzymatic activity and the mobility of cells after inhibition of MMP-9 gene expression.Material and Methods: The oligonucleotide shRNA insert had been designed to silence MMP-9 gene expression and was cloned into the pSUPER.neo expression vector. The construct was introduced into the HeLa (CCL-2 cervical cancer cells by lipotransfection. Simultaneously in control cells MMP-9 were inhibited by doxycycline. Changes in activity of MMP-9 were analyzed by gelatin zymography and wound-healing assay.Results/Conclusions: Gelatin zymography allowed us to confirm that activity of MMP-9 in cells transfected by shRNA-MMP-9 and treated by doxycycline were similar and significantly lower in comparison with control cells. Phenotypic tests of migration in vitro confirm statistically significant (P<0.05 changes in cell migration – control cells healed 3 to 5 times faster in comparison with transfected or doxycycline treated cells. Our studies show the significant role of MMP-9 in mobility and invasiveness of tumor cells, thus indicating a potential target point of interest for gene therapy.

  10. Prenatal nicotine increases matrix metalloproteinase 2 (MMP-2) expression in fetal guinea pig hearts.

    Science.gov (United States)

    Thompson, Loren P; Liu, Hongshan; Evans, LaShauna; Mong, Jessica A

    2011-11-01

    This study tested the hypothesis that maternal nicotine ingestion increases matrix metalloproteinase (MMP) expression in fetal hearts, which is mediated by the generation of reactive oxygen species. Timed pregnant guinea pigs were administered either water alone, nicotine (200 μg/mL), N-acetylcysteine (NAC), or nicotine plus NAC in their drinking water for 10 days at 52-day gestation (term = 65 days). Near-term (62 days), anesthetized fetuses were extracted, hearts were excised, and left cardiac ventricles snap frozen for analysis of MMP-2/-9/-13 protein and activity levels. Interstitial collagens were identified by Picrosirius red stain to assess changes in the extracellular matrix. Prenatal nicotine increased active MMP-2 forms and interstitial collagen but had no effect on either pro- or active MMP-9 or MMP-13 forms. In the presence of nicotine, NAC decreased active MMP-2 protein levels and reversed the nicotine-induced increase in collagen staining. We conclude that prenatal nicotine alters MMP-2 expression in fetal hearts that may be mediated by reactive oxygen species generation.

  11. Tissue levels of matrix metalloproteinases MMP-2 and MMP-9 are related to the overall survival of patients with gastric carcinoma

    NARCIS (Netherlands)

    Sier, C.F.M.; Kubben, F.J.G.M.; Ganesh, S.; Heerding, M.M.; Griffioen, G.; Hanemaaijer, R.; Krieken, J.H.J.M. van; Lamers, C.B.H.W.; Verspaget, H.W.

    1996-01-01

    Proteinases are involved in tumour invasion and metastasis. Several matrix metalloproteinases (MMPs) have been shown to be increased in various human carcinomas. We assessed the levels of MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in 50 gastric carcinomas and corresponding mucosa using

  12. Membrane-type-3 matrix metalloproteinase (MT3-MMP functions as a matrix composition-dependent effector of melanoma cell invasion.

    Directory of Open Access Journals (Sweden)

    Olga Tatti

    Full Text Available In primary human melanoma, the membrane-type matrix metalloproteinase, MT3-MMP, is overexpressed in the most aggressive nodular-type tumors. Unlike MT1-MMP and MT2-MMP, which promote cell invasion through basement membranes and collagen type I-rich tissues, the function of MT3-MMP in tumor progression remains unclear. Here, we demonstrate that MT3-MMP inhibits MT1-MMP-driven melanoma cell invasion in three-dimensional collagen, while yielding an altered, yet MT1-MMP-dependent, form of expansive growth behavior that phenocopies the formation of nodular cell colonies. In melanoma cell lines originating from advanced primary or metastatic lesions, endogenous MT3-MMP expression was associated with limited collagen-invasive potential. In the cell lines with highest MT3-MMP expression relative to MT1-MMP, collagen-invasive activity was increased following stable MT3-MMP gene silencing. Consistently, MT3-MMP overexpression in cells derived from less advanced superficially spreading melanoma lesions, or in the MT3-MMP knockdown cells, reduced MT1-MMP-dependent collagen invasion. Rather than altering MT1-MMP transcription, MT3-MMP interacted with MT1-MMP in membrane complexes and reduced its cell surface expression. By contrast, as a potent fibrinolytic enzyme, MT3-MMP induced efficient invasion of the cells in fibrin, a provisional matrix component frequently found at tumor-host tissue interfaces and perivascular spaces of melanoma. Since MT3-MMP was significantly upregulated in biopsies of human melanoma metastases, these results identify MT3-MMP as a matrix-dependent modifier of the invasive tumor cell functions during melanoma progression.

  13. Detection of the matrix metalloproteinases MMP-2 and MMP-9 and tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 in llama (Lama glama) oviduct.

    Science.gov (United States)

    Zampini, R; Argañaraz, M E; Miceli, D C; Apichela, S A

    2014-06-01

    Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are involved in several reproductive events like oocyte-spermatozoa interaction and semen liquefaction. In order to study their role in the llama oviductal reproductive process, MMP activity in oviductal fluid (OF) was assayed. Considering that llama genome sequences are partially known, a strategy to procure cDNA sequences of MMP-2, MMP-9, TIMP-1 and TIMP-2 was designed. Afterwards, their expression patterns in the different llama oviductal segments were assayed. Gelatine zymograms detected 62 and 94 kDa protease activities that matched MMP-2 and pro-MMP-9, respectively. Expression pattern analysis showed that MMP and TIMP mRNAs were present in ampulla, isthmus, utero-tubal junction (UTJ) and papilla. Altogether, these findings support the argument that MMPs/TIMPs are produced in the oviduct and secreted into the oviductal lumen. Our results encourage further studies to elucidate the role of these proteins in reproductive oviductal events.

  14. Identification of collagen binding domain residues that govern catalytic activities of matrix metalloproteinase-2 (MMP-2).

    Science.gov (United States)

    Mikhailova, Margarita; Xu, Xiaoping; Robichaud, Trista K; Pal, Sanjay; Fields, Gregg B; Steffensen, Bjorn

    2012-01-01

    An innovative approach to enhance the selectivity of matrix metalloproteinase (MMP) inhibitors comprises targeting these inhibitors to catalytically required substrate binding sites (exosites) that are located outside the catalytic cleft. In MMP-2, positioning of collagen substrate molecules occurs via a unique fibronectin-like domain (CBD) that contains three distinct modular collagen binding sites. To characterize the contributions of these exosites to gelatinolysis by MMP-2, seven MMP-2 variants were generated with single, or concurrent double and triple alanine substitutions in the three fibronectin type II modules of the CBD. Circular dichroism spectroscopy verified that recombinant MMP-2 wild-type (WT) and variants had the same fold. Moreover, the MMP-2 WT and variants had the same activity on a short FRET peptide substrate that is hydrolyzed independently of CBD binding. Among single-point variants, substitution in the module 3 binding site had greatest impact on the affinity of MMP-2 for gelatin. Simultaneous substitutions in two or three CBD modules further reduced gelatin binding. The rates of gelatinolysis of MMP-2 variants were reduced by 20-40% following single-point substitutions, by 60-75% after double-point modifications, and by >90% for triple-point variants. Intriguingly, the three CBD modules contributed differentially to cleavage of dissociated α-1(I) and α-2(I) collagen chains. Importantly, kinetic analyses (k(cat)/K(m)) revealed that catalysis of a triple-helical FRET peptide substrate by MMP-2 relied primarily on the module 3 binding site. Thus, we have identified three collagen binding site residues that are essential for gelatinolysis and constitute promising targets for selective inhibition of MMP-2.

  15. Pentosan polysulfate decreases prostate smooth muscle proliferation and extracellular matrix turnover.

    Science.gov (United States)

    Elliot, S J; Zorn, B H; McLeod, D G; Moul, J W; Nyberg, L; Striker, L J; Striker, G E

    2003-01-01

    Benign prostatic hyperplasia (BPH) involves proliferation of smooth muscle cells and increased deposition of extracellular matrix (ECM). We recently found that pentosan polysulfate (PPS) has marked effects on growth and ECM of smooth muscle cells derived from vascular tissues. We examined smooth muscle cells cultured from human prostates and the effects of PPS on their growth and ECM production. Fragments of surgical prostatectomy specimens were diced, digested with collagenase (0.01%), and placed in culture medium supplemented with 20% fetal bovine serum. Outgrowths of elongated cells were characterized by light microscopic examination and immunohistochemical techniques by the presence of F-actin, alpha-smooth muscle actin, and myosin, which is a characteristic of smooth muscle cells. Two independent isolates were propagated, and growth curves and ECM production were assessed in the presence and absence of PPS (10 or 100 microg/ml). PPS decreased cell number beginning at day 1 and throughout the incubation period, up to 4 days. The amount of the ECM degradative enzymes, metallo-proteinases MMP-9 and MMP-2, was examined by zymography. PPS did not alter the amount of MMP-2 in the supernatants but MMP-9 was increased 234.4 +/- 17.23-fold over control cells. Tissue inhibitor of MMP (TIMPS), examined by reverse zymography, increased 200% over control. The amount of alpha I type (IV) and alpha I type (I) collagen released in the supernatant, measured by ELISA, significantly decreased in PPS-treated cultures. In conclusion, we found that the administration of PPS decreased proliferation as well as ECM production in prostate smooth muscle. Since smooth muscle proliferation and ECM are involved in the pathophysiology of BPH, PPS may have therapeutic potential.

  16. The relationship between the first episode of wheezing and matrix metalloproteinases-9 and MMP-2 and tissue inhibitors of MMP-1 levels in preterm infants

    Directory of Open Access Journals (Sweden)

    Rabia Gonul Sezer

    2013-01-01

    Full Text Available Aims: Matrix metalloproteinases (MMP have been associated with neonatal lung morbidity and MMP dysregulation contributes to the pathology of chronic and acute lung disorders. Most of the previous studies were performed in the 1 st weeks of life of the preterm newborns. There are no data on the serum levels of MMP-2, MMP-9 or tissue inhibitors of matrix metalloproteinases (TIMP-1 from preterm infants recovering from lung morbidities. We aimed to compare MMP-2, MMP-9 and TIMP-1 levels in preterm and term infants hospitalized with their first episode of wheezing. Methods: We prospectively evaluated 18 preterm infants with a history of chronic lung disease, respiratory distress syndrome or oxygen therapy and 14 age- and sex-matched term infants who were admitted for a first episode of wheezing. We quantified total serum concentrations of MMP-2, MMP-9 and TIMP-1 to assess whether these serum markers levels were associated with the first episode of wheezing in infants with a history of oxygen therapy during the neonatal period. Results: Upon hospitalization, MMP-2 and TIMP-1 levels were higher in preterm infants than in term infants. In contrast, there was no significant relationship between MMP-9 levels or the MMP-9/TIMP-1 ratio between preterm and term infants. The area under the receiver operating characteristic curve for MMP-2 was 0.70 (95% confidence interval [CI] 0.51-0.89. The area under the curve for TIMP-1 was 0.78 (95% CI 0.61-0.94. MMP-9, MMP-2 and TIMP-1 levels did not correlate with gestational age, gender or severity of wheezing. Conclusion: The negative proportion of MMP-9 to TIMP-1 that we detected in term infants was not present in preterm infants. The balance of MMP-9 to TIMP-1 may have been disrupted by lung damage in the premature infants. Overproduction of MMP-2 and TIMP-1 in the serum may be associated with the pathogenesis of wheezing in preterm infants.

  17. Histochemical evidence of osteoclastic degradation of extracellular matrix in osteolytic metastasis originating from human lung small carcinoma (SBC-5) cells.

    Science.gov (United States)

    Li, Minqi; Amizuka, Norio; Takeuchi, Kiichi; Freitas, Paulo H L; Kawano, Yoshiro; Hoshino, Masaaki; Oda, Kimimitsu; Nozawa-Inoue, Kayoko; Maeda, Takeyasu

    2006-02-01

    The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)-positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular

  18. Induction of tissue inhibitor of matrix metalloproteinase-2 by cholesterol depletion leads to the conversion of proMMP-2 into active MMP-2 in human dermal fibroblasts.

    Science.gov (United States)

    Kim, Sangmin; Oh, Jang-Hee; Lee, Youngae; Lee, Jeongyoon; Cho, Kwang Hyun; Chung, Jin Ho

    2010-01-31

    Cholesterol is one of major components of cell membrane and plays a role in vesicular trafficking and cellular signaling. We investigated the effects of cholesterol on matrix metalloproteinase-2 (MMP-2) activation in human dermal fibroblasts. We found that tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) expression and active form MMP-2 (64 kD) were dose-dependently increased by methyl-beta-cyclodextrin (MbetaCD), a cholesterol depletion agent. In contrast, cholesterol depletion-induced TIMP-2 expression and MMP-2 activation were suppressed by cholesterol repletion. Then we investigated the regulatory mechanism of TIMP-2 expression by cholesterol depletion. We found that the phosphorylation of JNK as well as ERK was significantly increased by cholesterol depletion. Moreover, cholesterol depletion-induced TIMP-2 expression and MMP-2 activation was significantly decreased by MEK inhibitor U0126, and JNK inhibitor SP600125, respectively. While a low dose of recombinant TIMP-2 (100 ng/ml) increased the level of active MMP-2 (64 kD), the high dose of TIMP-2 (>or=200 ng/ml) decreased the level of active MMP-2 (64 kD). Taken together, we suggest that the induction of TIMP-2 by cholesterol depletion leads to the conversion of proMMP-2 (72 kD) into active MMP-2 (64 kD) in human dermal fibroblasts.

  19. Expression of matrix metalloproteinase-1 and tumor necrosis factor-α in ulcerative colitis

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To examine the expression of matrix metallo-proteinase-1 (MMP-1) and tumor necrosis factor-α (TNF-α) in the colon mucosa of patients with ulcerative colitis (UC).METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to examine the expression of MMP-1 and TNF-α at both mRNA and protein levels in the colon mucosa of patients with UC. Correlation between MMP-1 and TNF-α and their correlation with the severity of the disease were also analyzed statistically.RESULTS: The expression of MMP-1 and TNF-α in the ulcerated and inflamed colon mucosa of patients with UC was significantly higher than that in the non-inflamed mucosa of normal controls at both mRNA and protein levels. Furthermore, the expression of MMP-1 and TNF-α in the ulcerated area was significantly higher than that in the inflamed area of patients with UC (0.9797 ± 0.1433 vs 0.6746 ± 0.0373, 0.8669 ± 0.0746 vs 0.5227 ± 0.0435, P < 0.05). There was no statistically significant difference in the non-inflamed area of normal controls. There was a significant correlation between MMP-1 and TNF-α expression (0.9797 ± 0.1433 vs 0.8669 ± 0.0746, P < 0.05), the correlating factor was 0.877. MMP-1 and TNF-α showed a significant correlation with the severity of the disease (0.0915 ± 0.0044 vs 0.0749 ± 0.0032 , 0.0932 ± 0.0019 vs 0.0724 ± 0.0043, P < 0.05), their correlating factors were 0.942 and 0.890, respectively.CONCLUSION: Excessively expressed MMP-1 directly damages the colon mucosa by degrading extracellular matrix (ECM) in patients with UC. While damaging colon mucosa, excessively expressed TNF-α stimulates MMPs secreting cells to produce more MMP-1 and aggravates the mucosa damage. MMP-1 promotes secretion of TNF-a in a positive feedback manner to cause further injury in the colon mucosa. MMP-1 and TNF-α correlate well with the severity of the disease, and therefore, can be used clinically as biological markers to judge the severity of UC.

  20. Circulating levels of matrix metalloproteinase-9 (MMP-9, neutrophil gelatinase-associated lipocalin (NGAL and their complex MMP-9/NGAL in breast cancer disease

    Directory of Open Access Journals (Sweden)

    Nonni Afroditi

    2009-11-01

    Full Text Available Abstract Background Recent evidence suggests that neutrophil gelatinase-associated lipocalin (NGAL expression is induced in many types of human cancer, while detection of its complex with matrix metalloproteinase-9 (MMP-9 is correlated with cancer disease status. We aim to evaluate the serum expression of MMP-9, NGAL and their complex (MMP-9/NGAL during the diagnostic work-up of women with breast abnormalities and investigate their correlation with disease severity. Methods The study included 113 women with non-palpable breast lesions undergoing vacuum-assisted breast biopsy for histological diagnosis, and 30 healthy women, which served as controls. Expression levels of MMP-9, NGAL and their complex MMP-9/NGAL were determined in peripheral blood samples with immunoenzymatic assays. Results Women with invasive ductal carcinoma exhibited significantly increased levels of MMP-9, NGAL and MMP-9/NGAL compared to healthy controls (MMP-9: p Conclusion These findings suggest that the serum measurement of MMP-9 and NGAL may be useful in non-invasively monitoring breast cancer progression, while supporting their potential role as early biomarkers of breast disease status.

  1. Neutrophil Gelatinase-Associated Lipocalin (NGAL, Pro-Matrix Metalloproteinase-9 (pro-MMP-9 and Their Complex Pro-MMP-9/NGAL in Leukaemias

    Directory of Open Access Journals (Sweden)

    Sandrine Bouchet

    2014-04-01

    Full Text Available Matrix metalloproteinase (MMP-9 and neutrophil gelatinase-associated lipocalin (NGAL have gained attention as cancer biomarkers. The inactive zymogen form of MMP-9 (pro-MMP-9 also exists as a disulphide-linked heterodimer bound to NGAL in humans. Leukaemias represent a heterogeneous group of neoplasms, which vary in their clinical behavior and pathophysiology. In this review, we summarize the current literature on the expression profiles of pro-MMP-9 and NGAL as prognostic factors in leukaemias. We also report the expression of the pro-MMP-9/NGAL complex in these diseases. We discuss the roles of (pro-MMP-9 (active and latent forms and NGAL in tumour development, and evaluate the mechanisms by which pro-MMP-9/NGAL may influence the actions of (pro-MMP-9 and NGAL in cancer. Emerging knowledge about the coexpression and the biology of (pro-MMP-9, NGAL and their complex in cancer including leukaemia may improve treatment outcomes.

  2. Role of salivary matrix metalloproteinase-8 (MMP-8) in chronic periodontitis diagnosis.

    Science.gov (United States)

    Gupta, Namita; Gupta, N D; Gupta, Akash; Khan, Saif; Bansal, Neha

    2015-03-01

    Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases (MMPs) secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tissue, leading to chronic periodontitis. This study aimed to correlate salivary levels of MMP-8 and periodontal parameters of chronic periodontitis to establish MMP-8 as a noninvasive marker for the early diagnosis of chronic periodontitis. The study involved 40 subjects visiting the periodontic OPD of Dr. Ziauddin Ahmad Dental College and Hospital, located in Aligarh, U.P., India, from 2011 to 2012. The subjects were divided into two groups: group I consisted of 20 periodontally healthy subjects (controls) while group II consisted of 20 patients with chronic periodontitis. Chronic periodontitis was assessed on the basis of several periodontal parameters, including pocket probing depth (PPD), clinical attachment level (CAL), gingival index (GI), and plaque index (PI). Around 3ml of unstimulated and whole expectorated saliva was collected for MMP-8 estimation by ELISA using Quantikine human total MMP-8 immunoassay kits. Data were analyzed using STATISTICA (Windows version 6) software. Salivary MMP-8 levels of groups I and II were 190.91 ± 143.89 ng/ml and 348.26 ± 202.1 ng/ml, respectively. The MMP-8 levels and periodontal status (PPD, CAL, GI, and PI) of groups I and II showed positive and significant correlations (for PPD, r = 0.63, P chronic periodontitis.

  3. Serum matrix metalloproteinase 9 (MMP9) as a biochemical marker for wasting marmoset syndrome.

    Science.gov (United States)

    Yoshimoto, Takuro; Niimi, Kimie; Takahashi, Eiki

    2016-06-01

    Use of the common marmoset (Callithrix jacchus) as a non-human primate experimental animal has increased in recent years. Although wasting marmoset syndrome (WMS) is one of the biggest problems in captive marmoset colonies, the molecular mechanisms, biochemical markers for accurate diagnosis and a reliable treatment remain unknown. In this study, as a first step to finding biochemical marker(s) for the accurate diagnosis of WMS, we conducted blood cell counts, including hematocrit, hemoglobin and platelets, and examined serum chemistry values, including albumin, calcium and levels of serum matrix metalloproteinase 9 (MMP9), using a colony of marmosets with and without weight loss. MMP9 is thought to be an enzyme responsible for the degradation of extracellular matrix components and participates in the pathogenesis of inflammatory conditions, such as human and murine inflammatory bowel disease, which, like WMS, are characterized histologically by inflammatory cell infiltrations in the intestines. The values of hematocrit and hemoglobin and levels of serum albumin and calcium in the WMS group were significantly decreased versus the control group. The platelet values and serum MMP9 concentrations were increased significantly in the WMS group compared with the control group. MMP9 could be a new and useful marker for the diagnosis of WMS in addition to hematocrit, hemoglobin, serum albumin and calcium. Our results also indicate that MMP9 could be a useful molecular candidate for treatment.

  4. Expressions of Matrix Metalloproteinases (MMP-2, MMP-7, and MMP-9 and Their Inhibitors (TIMP-1, TIMP-2 in Inflammatory Bowel Diseases

    Directory of Open Access Journals (Sweden)

    Katarzyna Jakubowska

    2016-01-01

    Full Text Available Crohn’s disease (CD and ulcerative colitis (UC belong to a group of inflammatory bowel diseases (IBD. The aim of our study was to evaluate the expression of MMP-2, MMP-7, MMP-9, TIMP-1, and TIMP-2 in ulcerative colitis and Crohn’s disease. The study group comprised 34 patients with UC and 10 patients with CD. Evaluation of MMP-2, MMP-7, MMP-9, TIMP-1, and TIMP-2 expression in tissue samples was performed using immunohistochemistry. The overexpression of MMP-9 and TIMP-1 was dominant in both the glandular epithelium and inflammatory infiltration in UC patients. In contrast, in CD subjects the positive expression of MMP-2 and TIMP-1 was in glandular tubes while mainly MMP-7 and TIMP-2 expression was in inflammatory infiltration. Metalloproteinases’ expression was associated with the presence of erosions, architectural tissue changes, and inflammatory infiltration in the lamina propria of UC patients. The expression of metalloproteinase inhibitors correlated with the presence of eosinophils and neutrophils in UC and granulomas in CD patients. Our studies indicate that the overexpression of metalloproteinases and weaker expression of their inhibitors may determine the development of IBD. It appears that MMP-2, MMP-7, and MMP-9 may be a potential therapeutic target and the use of their inhibitors may significantly reduce UC progression.

  5. 乳腺癌组织中 VEGF-D、MMP-2及 MMP-9的表达及其临床意义%The Expressions and Clinical Significance of Matrix Metalloproteinases 2(MMP-2),Ma-trix Metalloproteinases 9 (MMP-9) and Vascular Endothelial Growth Factor-D (VEGF-D) in Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    张阿娜

    2016-01-01

    目的:探讨乳腺癌组织中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)和血管内皮生长因子D( VEGF-D)表达的临床意义。方法采用免疫组化法,对60例乳腺癌组织及60例正常组织中 MMP-2、MMP-9及VEGF-D表达情况进行检测。结果60例乳腺癌组织中,MMP-2、MMP-9阳性表达率分别为61.67%、56.67%,VEGF-D阳性表达率为61.67%。 MMP-2、MMP-9及VEGF-D阳性表达率Ⅲ~Ⅳ期者显著高于Ⅰ~Ⅱ期者;低分化者显著高于高、中分化者;有淋巴结转移者显著高于无淋巴结转移者;且MMP-2与VEGF-D表达,MMP-9与VEGF-D表达均呈正相关性。结论低分化、Ⅲ~Ⅳ期乳腺癌患者高表达MMP-2、MMP-9与VEGF-D,MMPs与VEGF-D表达呈正相关性,可通过阻断VEGF-D及MMPs活性控制、治疗乳腺癌。%Objective To explore the expressions of matrix metalloproteinases 2 ( MMP-2 ) ,matrix metalloproteinases 9 (MMP-9) and vascular endothelial growth factor-D(VEGF-D)and their significance in breast cancer tissues.Methods The ex-pressions of MMP-2,MMP-9 and VEGF-D in 60 cases of breast cancer tissues and 60 cases of normal esophageal tissues were de-tected by immunohistochemistry.Results The expression rates of MMP-2,MMP-9 and VEGF-D in breast cancer tissues were 61.67%,56.67%and 61.67%.The expression rates of MMP-2,MMP-9 and VEGF-D in grade Ⅲ-Ⅳ were significantly higher than that of gradeⅠ-Ⅱ.The expression rates of MMP-2,MMP-9 and VEGF-D in patients with poor differentiation were signifi-cantly higher than that with well-mediate differentiation.The expression rates of MMP-2,MMP-9 and VEGF-D in patients with lymph node metastasis were significantly higher than that without lymph node metastasis.MMP-2 and VEGF-D expression were positively correlated.MMP-9 and VEGF-D expression were also positively correlated.Conclusion The expression rates of MMP-2,MMP-9 and VEGF-D are closely related to poor differentiation

  6. Intracellular matrix metalloproteinase-2 (MMP-2) regulates human platelet activation via hydrolysis of talin.

    Science.gov (United States)

    Soslau, Gerald; Mason, Christopher; Lynch, Stephen; Benjamin, James; Ashak, Dani; Prakash, Jamunabai M; Moore, Andrew; Bagsiyao, Pamela; Albert, Trevine; Mathew, Lynn M; Jost, Monika

    2014-01-01

    Matrix metalloproteinase (MMP) activity is generally associated with normal or pathological extracellular processes such as tissue remodelling in growth and development or in tumor metastasis and angiogenesis. Platelets contain at least three MMPs, 1, 2 and 9 that have been reported to stimulate or inhibit agonist-induced platelet aggregation via extracellular signals. The non-selective Zn+2 chelating MMP inhibitor, 1,10-phenanthroline, and the serine protease inhibitor, AEBSF, were found to inhibit all tested agonist-induced platelet aggregation reactions. In vitro analysis demonstrated that 1,10-phenanthroline completely inhibited MMP-1,2,and 9 but had little to no effect on calpain activity while the converse was true with AEBSF. We now demonstrate that MMP-2 functions intracellularly to regulate agonist-induced platelet aggregations via the hydrolytic activation of talin, the presumed final activating factor of glycoprotein (GP)IIb/IIIa integrin (the inside-out signal). Once activated GPIIb/IIIa binds the dimeric fibrinogen molecule required for platelet aggregation. The active intracellular MMP-2 molecule is complexed with JAK 2/STAT 3, as demonstrated by the fact that all three proteins are co-immunoprecipitated with either anti-JAK 2, or anti-STAT 3 antibodies and by immunofluorescence studies. The MMP-2 platelet activation pathway can be synergistically inhibited with the non-selective MMP inhibitor, 1,10-phenanthroline, plus a JAK 2 inhibitor. This activation pathway is distinct from the previously reported calpain-talin activating pathway. The identification of a new central pathway for platelet aggregation presents new potential targets for drug regulation and furthers our understanding of the complexity of platelet activation mechanisms.

  7. A self-propagating matrix metalloprotease-9 (MMP-9 dependent cycle of chronic neutrophilic inflammation.

    Directory of Open Access Journals (Sweden)

    Xin Xu

    Full Text Available BACKGROUND: Chronic neutrophilic inflammation is a poorly understood feature in a variety of diseases with notable worldwide morbidity and mortality. We have recently characterized N-acetyl Pro-Gly-Pro (Ac-PGP as an important neutrophil (PMN chemoattractant in chronic inflammation generated from the breakdown of collagen by the actions of MMP-9. MMP-9 is present in the granules of PMNs and is differentially released during inflammation but whether Ac-PGP contributes to this ongoing proteolytic activity in chronic neutrophilic inflammation is currently unknown. METHODOLOGY/PRINCIPAL FINDINGS: Utilizing isolated primary blood PMNs from human donors, we found that Ac-PGP induces significant release of MMP-9 and concurrently activates the ERK1/2 MAPK pathway. This MMP-9 release is attenuated by an inhibitor of ERK1/2 MAPK and upstream blockade of CXCR1 and CXCR2 receptors with repertaxin leads to decreased MMP-9 release and ERK 1/2 MAPK activation. Supernatants obtained from PMNs stimulated by Ac-PGP generate more Ac-PGP when incubated with intact collagen ex vivo; this effect is inhibited by an ERK1/2 pathway inhibitor. Finally, clinical samples from individuals with CF demonstrate a notable correlation between Ac-PGP (as measured by liquid chromatography-tandem mass spectrometry and MMP-9 levels even when accounting for total PMN burden. CONCLUSIONS/SIGNIFICANCE: These data indicate that ECM-derived Ac-PGP could result in a feed-forward cycle by releasing MMP-9 from activated PMNs through the ligation of CXCR1 and CXCR2 and subsequent activation of the ERK1/2 MAPK, highlighting for the first time a matrix-derived chemokine (matrikine augmenting its generation through a discrete receptor/intracellular signaling pathway. These findings have notable implications to the development unrelenting chronic PMN inflammation in human disease.

  8. Domain structure and function of matrix metalloprotease 23 (MMP23): role in potassium channel trafficking.

    Science.gov (United States)

    Galea, Charles A; Nguyen, Hai M; George Chandy, K; Smith, Brian J; Norton, Raymond S

    2014-04-01

    MMP23 is a member of the matrix metalloprotease family of zinc- and calcium-dependent endopeptidases, which are involved in a wide variety of cellular functions. Its catalytic domain displays a high degree of structural homology with those of other metalloproteases, but its atypical domain architecture suggests that it may possess unique functional properties. The N-terminal MMP23 pro-domain contains a type-II transmembrane domain that anchors the protein to the plasma membrane and lacks the cysteine-switch motif that is required to maintain other MMPs in a latent state during passage to the cell surface. Instead of the C-terminal hemopexin domain common to other MMPs, MMP23 contains a small toxin-like domain (TxD) and an immunoglobulin-like cell adhesion molecule (IgCAM) domain. The MMP23 pro-domain can trap Kv1.3 but not closely-related Kv1.2 channels in the endoplasmic reticulum, preventing their passage to the cell surface, while the TxD can bind to the channel pore and block the passage of potassium ions. The MMP23 C-terminal IgCAM domain displays some similarity to Ig-like C2-type domains found in IgCAMs of the immunoglobulin superfamily, which are known to mediate protein-protein and protein-lipid interactions. MMP23 and Kv1.3 are co-expressed in a variety of tissues and together are implicated in diseases including cancer and inflammatory disorders. Further studies are required to elucidate the mechanism of action of this unique member of the MMP family.

  9. Matrix metalloproteinase 1 (MMP1) is associated with early-onset lung cancer.

    Science.gov (United States)

    Sauter, Wiebke; Rosenberger, Albert; Beckmann, Lars; Kropp, Silke; Mittelstrass, Kirstin; Timofeeva, Maria; Wölke, Gabi; Steinwachs, Angelika; Scheiner, Daniela; Meese, Eckart; Sybrecht, Gerhard; Kronenberg, Florian; Dienemann, Hendrik; Chang-Claude, Jenny; Illig, Thomas; Wichmann, Heinz-Erich; Bickeböller, Heike; Risch, Angela

    2008-05-01

    Matrix metalloproteinases (MMP) play a key role in the breakdown of extracellular matrix and in inflammatory processes. MMP1 is the most highly expressed interstitial collagenase degrading fibrillar collagens. Overexpression of MMP1 has been shown in tumor tissues and has been suggested to be associated with tumor invasion and metastasis. Nine haplotype tagging and additional two intronic single nucleotide polymorphisms (SNP) of MMP1 were genotyped in a case control sample, consisting of 635 lung cancer cases with onset of disease below 51 years of age and 1,300 age- and sex-matched cancer-free controls. Two regions of linkage disequilibrium (LD) of MMP1 could be observed: a region of low LD comprising the 5' region including the promoter and a region of high LD starting from exon 1 to the end of the gene and including the 3' flanking region. Several SNPs were identified to be individually significantly associated with risk of early-onset lung cancer. The most significant effect was seen for rs1938901 (P = 0.0089), rs193008 (P = 0.0108), and rs996999 (P = 0.0459). For rs996999, significance vanished after correction for multiple testing. For each of these SNPs, the major allele was associated with an increase in risk with an odds ratio between 1.2 and 1.3 (95% confidence interval, 1.0-1.5). The haplotype analysis supported these findings, especially for subgroups with high smoking intensity. In summary, we identified MMP1 to be associated with an increased risk for lung cancer, which was modified by smoking.

  10. Site controlled transgenic mice validating increased expression from human matrix metalloproteinase (MMP-1) promoter due to a naturally occurring SNP.

    Science.gov (United States)

    Coon, Charles I; Fiering, Steven; Gaudet, Justin; Wyatt, Colby A; Brinckerhoff, Constance E

    2009-09-01

    Matrix metalloproteinases (MMPs) comprise a family of more than 20 members, each with the ability to degrade components of the extracellular matrix. The interstitial collagenases have the unique capacity to degrade the stromal collagens, types I, II and III, the body's most abundant proteins. These collagenases include MMP-1, MMP-8, MMP-13 and MMP-14. MMP-1, with a very broad expression pattern, has major roles in mediating matrix destruction in many diseases. We have described a single nucleotide polymorphism (SNP) in the MMP-1 promoter that augments transcription. This SNP is the presence or absence of an extra guanine (G) at -1607 bp, which creates the sequence 5'-GGAA-3'(2G allele), and which is an ETS binding site. Compared to the 1G allele (5'-GAA-3'), the 2G SNP is associated with enhanced transcription of MMP-1 and increased enzymatic activity. Although murine systems are often used to model human diseases, mice have only distant homologues of human MMP-1. Therefore, we used a technique for the targeted insertion of a single copy of a gene at the HPRT locus to compare expression of the 1G and 2G alleles. We generated transgenic mice with -4372 bp of the human MMP-1 promoter containing either the 1G or 2G SNP in front of the lac Z (E.coli ss-galactosidase) gene. We measured the relative expression of the transgenes in vitro in embryonic stem (ES) cells and in fibroblasts derived from embryonic mice. Our data show modest constitutive expression of ss-galactosidase mRNA and protein from these alleles, with the 2G allele more transcriptionally active than the 1G allele. We conclude that these mice represent a model for integration of a single copy of the human MMP-1 promoter into the murine genome, and could be used to study MMP-1 gene expression in a murine system.

  11. Selective gene transfer to endometrial cancer cells by a polymer against matrix metalloproteinase 2 (MMP-2).

    Science.gov (United States)

    Han, Joo Youn; Choi, Dong Soon; Kim, Changhoon; Joo, Hyun; Min, Churl K

    2008-04-01

    A novel cancer-cell-specific gene delivery vector with high transfection efficiency was designed and tested with an in vitro coculture consisting of the human endometrial adenocarcinoma cell line, HEC-1A cells, and normal endometrial stromal cells. For the cancer-cell targeting, polyethylenimine (PEI), a cationic polymer that can be easily combined with anionic DNA to form a particulate complex, polyplex, being capable of transferring a gene into a variety of cells, was covalently conjugated with antibodies against matrix metalloproteinase 2 (MMP-2), a typical surface-marker protein on cancer cells known for its close correlation with angiogenesis and invasion in many types of cancer, using the heterofunctional cross-linker, n-succinimidyl 3-(2-pyridyldithio)-propionamide. Biophysical properties and transfection efficiencies of anti-MMP-2-conjugated PEI were analyzed by means of dynamic light scattering, laser Doppler anemometry, and flow cytometry. Our results reveal that (1) the PEI-anti-MMP-2 antibody conjugate maintains physical parameters, including sizes and surface charges, which appear to be favorable for gene transfer and (2) when the pEGFP-N3 plasmid complexes of the PEI-anti-MMP-2 antibody conjugate are applied to the coculture consisting of HEC-1A cells and human stromal cells, a high level of green fluorescent protein expression occurs in HEC-1A cells over stromal cells, suggesting a specific gene transfer targeting cancer cells. Therefore, targeting invading cancer cells with the PEI-anti-MMP-2 antibody conjugate could be promising in endometrial cancer treatment, and this gene delivery system deserves further optimization in the context of targeted therapeutic gene delivery.

  12. Tumor necrosis factor-α-accelerated degradation of type I collagen in human skin is associated with elevated matrix metalloproteinase (MMP)-1 and MMP-3 ex vivo

    DEFF Research Database (Denmark)

    Ågren, Magnus S; Schnabel, Reinhild; Christensen, Lise H

    2015-01-01

    /ml) in the absence or presence of the nonselective MMP inhibitor GM6001 for 8 days. The basal culture conditions promoted type I collagen catabolism that was accelerated by TNF-α (p...Tumor necrosis factor (TNF)-α induces matrix metalloproteinases (MMPs) that may disrupt skin integrity. We have investigated the effects and mechanisms of exogenous TNF-α on collagen degradation by incubating human skin explants in defined serum-free media with or without TNF-α (10ng...... were associated with increased collagen degradation. TNF-α increased secretion of MMP-1 (p

  13. Altered distribution of extracellular matrix proteins in the periodontal ligament of periostin-deficient mice.

    Science.gov (United States)

    Tabata, Chihiro; Hongo, Hiromi; Sasaki, Muneteru; Hasegawa, Tomoka; de Freitas, Paulo Henrique Luiz; Yamada, Tamaki; Yamamoto, Tomomaya; Suzuki, Reiko; Yamamoto, Tsuneyuki; Oda, Kimimitsu; Li, Minqi; Kudo, Akira; Iida, Junichiro; Amizuka, Norio

    2014-06-01

    Verifying whether periostin affects the distribution of type I collagen, fibronectin and tenascin C in the periodontal ligament (PDL) is important to contribute to a more thorough understanding of that protein's functions. In this study, we have histologically examined incisor PDL of mandibles in 20 week-old male wild-type and periostin-deficient (periostin-/-) mice, by means of type I collagen, fibronectin, tenascin C, proliferating cell nuclear antigen, matrix metallo-proteinase (MMP)-1 and F4/80-positive monocyte/macrophage immunostaining, transmission electron microscopy and quantitative analysis of cell proliferation. Wild-type PDL featured well-arranged layers of collagen bundles intertwined with PDL cells, whose longitudinal axis ran parallel to the collagen fibers. However, cells in the periostin-/- PDL were irregularly distributed among collagen fibrils, which were also haphazardly arranged. Type I collagen and fibronectin reactivity was seen throughout the wild-type PDL, while in the periostin-/- PDL, only focal, uneven staining for these proteins could be seen. Similarly, tenascin C staining was evenly distributed in the wild-type PDL, but hardly seen in the periostin-/- PDL. MMP-1 immunoreactivity was uniformly distributed in the wild-type PDL, but only dotted staining could be discerned in the periostin-/- PDL. F4/80-positive monocyte/macrophages were found midway between tooth- and bone-related regions in the wild-type PDL, a pattern that could not be observed in the periostin-/- PDL. In summary, periostin deficiency may not only cause PDL collagen fibril disorganization, but could also affect the distribution of other major extracellular matrix proteins such as fibronectin and tenascin C.

  14. Localization and possible role of membrane type metallo-proteinase and tissue inhibitors of metalloproteinase-1 in early stages of placentation

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Human placental tissues from the first and second trimesters of gestation have been investigated using riboprobe in situ hybridisation of mRNA sequences coding for membrane type metalloproteinase (MT-1-MMP) and tissue inhibitors of metalloproteinase-1 (TIMP-1). Results show that (i) both mRNAs express at a relatively high level in the chorion laeve trophoblast cells and the adjacent decidual cells of fetal membrane; (ii) the most abundant expression of the two mRNAs was found in the extravillous trophoblast between Rohrs and Nitabuch striae of basal plate, trophoblast shell and gland cells of the decidua; (iii) isolated or small groups of cytotrophoblast cells in the chorionic villi and in the cells lining arterioles in decidua and stem villi also expressed both MT-1-MMP and TIMP-1 at defferent extents. The data suggest that the coordinated expression of the MT-MMP and its inhibitor TIMP in defferent cells of the placental tissue may play an essential role in trophoblast invasion and angiogenesis related to placentation in the first two trimesters of gestation. They may also have an ability to effect separation of fetal from material tissue at a favorable junctional site during parturition.

  15. Matrix metalloproteinase 9 (MMP-9) in osteosarcoma: review and meta-analysis.

    Science.gov (United States)

    Wang, Jing; Shi, Qiong; Yuan, Tai-Xian; Song, Qi-Lin; Zhang, Yan; Wei, Qiang; Zhou, Lan; Luo, Jinyong; Zuo, Guowei; Tang, Min; He, Tong-Chuan; Weng, Yaguang

    2014-06-10

    The aim of this study is to determine the value of matrix metalloproteinase 9 (MMP-9) in diagnosis of osteosarcoma (OS). A systematic review and meta-analysis was conducted using MEDLINE, Embase, ISI Web of Knowledge, the Cochrane Library, Scopus, BioMed Central, ScienceDirect, China Biomedical literature Database (CBM) and China National Knowledge Internet (CNKI) from inception through Aug 29, 2013. Articles written in English or Chinese that investigated the accuracy of MMP-9 for the diagnosis of OS were included. Pooled sensitivity, specificity and the area under the receiver operating characteristic curve (AUC) were determined. I(2) was used to test heterogeneity and source of heterogeneity was investigated by meta-regression (tested with Meta-DiSc and STATA 12.0 statistical softwares). A total of 3729 articles were retrieved, of which 18 were included, accounting for 892 patients. Overall, the pooled sensitivity, specificity and AUC were 0.78 (95% CI 0.730-0.83), 0.90 (95% CI 0.79-0.95), and 0.87 (95% CI 0.83-0.89), respectively. The studies had substantial heterogeneity (I(2)=84%, 95% CI 65-100) (96%, 95% CI 94-99). Assay kit subgroup was the main source of the heterogeneity. Although MMP-9 was identified as a potential biomarker for OS, more studies were clearly needed to establish its diagnostic value.

  16. Epilysin (MMP-28) is deposited to the basolateral extracellular matrix of epithelial cells.

    Science.gov (United States)

    Heiskanen, Tuomas J; Illman, Sara A; Lohi, Jouko; Keski-Oja, Jorma

    2009-03-01

    Epilysin (MMP-28) is a conserved member of the matrix metalloproteinase (MMP) family. It is expressed in various normal tissues, and induced in wounds and in developing and regenerating nerves. Epilysin induces TGF-beta mediated epithelial to mesenchymal transition, but its other functions are largely unknown. We have characterized the localization of both catalytically active and mutated inactive, overexpressed epilysin in established epithelial cell lines. We found that epilysin was localized abundantly to the basolateral side of the cells and associated with the extracellular matrix (ECM) as verified by immunoblotting and confocal microscopy. Overexpression of epilysin in MDCK cells resulted in a drastic reduction of basolateral ECM, as observed by the disappearance of collagen type IV, laminin and fibronectin. Cultivation of epilysin expressing MDCK cells in defined serum free medium resulted in the restoration of these proteins to the ECM. The levels of fibronectin and collagen IV were, however, reduced in epilysin expressing cells under the serum free conditions, and degradation fragments of collagen IV were detected supporting the activation of proteolysis by epilysin. Epilysin was observed in its unprocessed 50 kDa active form in the ECM of MDCK cells under serum free conditions whereas in cells cultured in serum containing it was processed to the 48 kDa form. Current results indicate that epilysin associates with the basolateral ECM of cultured epithelial cells, where it plausibly plays a role in the regulation of matrix composition and turnover.

  17. IL-21 R、MMP-2在非小细胞肺癌中的表达及相关性研究%Correlation study of expression of IL-21 R and MMP-2 in patients with non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    戚胜波; 于奇

    2014-01-01

    目的:检测白介素-21受体(interleukin-21R,IL-21R)、基质金属蛋白酶-2(matrix metalloprotein-ases-2,MMP-2)在非小细胞肺癌( Non-small-cell lung cancer,NSCLC)癌组织中的表达,并探讨IL-21R、MMP-2的表达在NSCLC发生、发展及浸润转移的影响。方法采用免疫组化法检测50例NSCLC组织及20例正常肺组织对照组中IL-21R、MMP-2的表达,分析其阳性表达率与NSCLC患者临床病理特征的关系,并探讨其相关性。结果 NSCLC癌组织中IL-21R、MMP-2的阳性表达率均显著高于正常肺组织,且两者表达呈负相关(P<0.05);IL-21R和MMP-2的表达与淋巴结转移、临床分期及组织分化程度相关。结论 NSCLC中MMP-2的表达与其浸润转移正相关,其表达增加表示NSCLC有较高恶性生物学行为;IL-21R的表达与NSCLC浸润转移的呈负相关,且IL-21R、MMP-2在NSCLC发生及浸润过程中具有相关性。%Objective To investigate the expression of interleukin-21 receptor (IL-2R) and matrix metallo-proteinase-2 (MMP-2) in patients with non-small cell lung cancer (NSCLC), and to discuss the influence of the ex-pression of IL-21R and MMP-2 to the disease. Methods The expression of IL-21R and MMP-2 was detected by im-munohistochemical method in 50 patients with non-small cell lung cancer and 20 people with normal lung tissue. The correlation of the positive expression rate of IL-21R and MMP-2 to pathological characteristics was analyzed. Results The positive rate of IL-2R and MMP-2 in NSCLC patients was significantly higher than in the control group, and they were negatively correlated (P<0. 05). The expression of IL-21R and MMP-2 was related to clinical stages, lymph node metastasis status and tissue differentiation degree. Conclusion The expression of MMP-2 in NSCLC pa-tients is positively related with metastasis, which the increased expression shows higher malignant biological behavior in NSCLC patients. The expression of IL-21 R shows a negative correlation

  18. Increasing extracellular matrix collagen level and MMP activity induces cyst development in polycystic kidney disease

    Directory of Open Access Journals (Sweden)

    Liu Bin

    2012-09-01

    Full Text Available Abstract Background Polycystic Kidney Disease (PKD kidneys exhibit increased extracellular matrix (ECM collagen expression and metalloproteinases (MMPs activity. We investigated the role of these increases on cystic disease progression in PKD kidneys. Methods We examined the role of type I collagen (collagen I and membrane bound type 1 MMP (MT1-MMP on cyst development using both in vitro 3 dimensional (3D collagen gel culture and in vivo PCK rat model of PKD. Results We found that collagen concentration is critical in controlling the morphogenesis of MDCK cells cultured in 3D gels. MDCK cells did not form 3D structures at collagen I concentrations lower than 1 mg/ml but began forming tubules when the concentration reaches 1 mg/ml. Significantly, these cells began to form cyst when collagen I concentration reached to 1.2 mg/ml, and the ratios of cyst to tubule structures increased as the collagen I concentration increased. These cells exclusively formed cyst structures at a collagen I concentration of 1.8 mg/ml or higher. Overexpression of MT1-MMP in MDCK cells significantly induced cyst growth in 3D collagen gel culture. Conversely, inhibition of MMPs activity with doxycycline, a FDA approved pan-MMPs inhibitor, dramatically slowed cyst growth. More importantly, the treatment of PCK rats with doxycycline significantly decreased renal tubule cell proliferation and markedly inhibited the cystic disease progression. Conclusions Our data suggest that increased collagen expression and MMP activity in PKD kidneys may induce cyst formation and expansion. Our findings also suggest that MMPs may serve as a therapeutic target for the treatment of human PKD.

  19. Peptide from the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) inhibits membrane activation of matrix metalloproteinase-2 (MMP-2).

    Science.gov (United States)

    Xu, Xiaoping; Mikhailova, Margarita; Chen, Zhihua; Pal, Sanjay; Robichaud, Trista K; Lafer, Eileen M; Baber, Sam; Steffensen, Bjorn

    2011-09-01

    Cellular activation of latent matrix metalloproteinase-2 (proMMP-2) requires formation of a cell membrane-associated activation complex that involves specific binding between the hemopexin domain of proMMP-2 (PEX) and the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (C-TIMP-2). In this study, we tested the feasibility of inhibiting activation of proMMP-2 by exogenous inhibitors, which block the binding between PEX and TIMP-2. The recombinant C-TIMP-2 and synthetic peptides from C-TIMP-2 were used as inhibitors for proMMP-2 activation. Recombinant C-TIMP-2 bound specifically to both the catalytically inactive MMP-2(E404A) and the C-terminal domain of MMP-2 (PEX) in a concentration dependent manner with apparent K(d) of 3.9×10(-7)M and 1.7×10(-7)M, respectively. Moreover, C-TIMP-2 competed the binding between MMP-2(E404A) and full-length TIMP-2. Finally, activity assays showed that addition of C-TIMP-2 to HT-1080 fibrosarcoma cells inhibited proMMP-2 activation in a concentration-dependent manner. We then designed a synthetic peptide, P175L, consisting of 20 residues from the PEX-binding tail region of C-TIMP-2. P175L bound PEX and inhibited cell membrane-mediated activation of proMMP-2 in a concentration dependent manner. Deletion of the last 9 tail residues of C-TIMP-2 in P175L abrogated the inhibitory activities of the peptide showing that these residues were essential for function. Overall, these experiments have demonstrated that proMMP-2 activation can be inhibited by exogenous inhibitors which points to a potential strategy for MMP-2 specific inhibition.

  20. Serum haptoglobin-matrix metalloproteinase 9 (Hp-MMP 9) complex as a biomarker of systemic inflammation in cattle.

    Science.gov (United States)

    Bannikov, G A; Hinds, C A; Rajala-Schultz, P J; Premanandan, C; Rings, D M; Lakritz, J

    2011-01-01

    A reliable and specific test that discriminates between acute neutrophil activation and chronic inflammatory disease may be useful in clinical decision making in a variety of conditions encountered in veterinary medical practice. An ELISA specific for neutrophil-derived haptoglobin-matrix metalloproteinase 9 (Hp-MMP 9) complexes was used to determine serum concentrations of Hp-MMP 9 and was compared to ELISA assays for Haptoglobin (Hp) and matrix metalloproteinase 9 (MMP 9) in 15 animals with acute sepsis, 10 animals with chronic inflammatory or metabolic disease and 10 healthy cows. Animal disease classifications were completed prior to the determination of serum concentrations of the 3 proteins. Duration of illness, disease process and lesions observed at necropsy were used to place animals into a specific classification. The serum MMP 9 concentrations in healthy cows differed significantly from those measured in sera of acutely septic and chronically ill animals. Serum haptoglobin concentrations in healthy cows were negligible when compared to animals with acute septic or chronic diseases. There was substantial overlap in MMP 9 and Hp concentrations between acute and chronic disease animals. In contrast, serum concentrations of Hp-MMP 9 complexes found almost exclusively in sera from acutely septic animals but not in chronically ill and normal cattle. The Hp-MMP 9 ELISA may be the serological test of choice in the determination of systemic inflammation associated with bacterial sepsis.

  1. Matrix metalloproteinase 2 fused to GFP, expressed in E. coli, successfully tracked MMP-2 distribution in vivo.

    Science.gov (United States)

    Azevedo, A; Prado, A F; Issa, J P M; Gerlach, R F

    2016-08-01

    Matrix Metalloproteinases (MMPs) participate in many physiological and pathological processes. One major limitation to a better understanding of the role MMPs play in these processes is the lack of well-characterized chimeric proteins and characterization of their fluorescence. The specialized literature has reported on few constructs bearing MMPs fused to the sequence of the green fluorescent protein (GFP), but none of the described constructs have been intended for expression in bacteria or for purification and use in vivo. This work has tested a recombinant reporter protein containing the MMP-2 catalytic domain fused to GFP in terms of purification efficiency, degradation of substrates in solution and in zymograms, kinetic activity, GFP fluorescence, and GFP fluorescence in whole animals after injection of the purified and lyophilized fluorescent protein. This work has also characterized rhMMP-2 (recombinant human MMP-2) and inactive clones and used them as negative controls in experiments employing catMMP-2/GFP and rhMMP-2. To our knowledge, this is the first study that has fully characterized a chimeric protein with the MMP-2 catalytic domain fused to GFP, that has efficiently purified such protein from bacteria in a single-step, and that has obtained an adequate chimeric protein for injection in animals and tracking of MMP-2 fate and activity in vivo.

  2. Imaging with radiolabelled anti-membrane type 1 matrix metalloproteinase (MT1-MMP) antibody: potentials for characterizing atherosclerotic plaques

    Energy Technology Data Exchange (ETDEWEB)

    Kuge, Yuji [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); Hokkaido University, Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Sapporo (Japan); Hokkaido University, Central Institute of Isotope Science, Sapporo (Japan); Takai, Nozomi; Ogawa, Yuki; Temma, Takashi; Nishigori, Kantaro; Ishino, Seigo; Kamihashi, Junko; Saji, Hideo [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); Zhao, Yan [Hokkaido University, Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Sapporo (Japan); Kiyono, Yasushi [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); University of Fukui, Biomedical Imaging Research Center, Fukui (Japan); Shiomi, Masashi [Kobe University Graduate School of Medicine, Institute for Experimental Animals, Kobe (Japan)

    2010-11-15

    Membrane type 1 matrix metalloproteinase (MT1-MMP) activates pro-MMP-2 and pro-MMP-13 to their active forms and plays important roles in the destabilization of atherosclerotic plaques. This study sought to determine the usefulness of {sup 99m}Tc-labelled monoclonal antibody (mAb), recognizing MT1-MMP, for imaging atherosclerosis in a rabbit model (WHHLMI rabbits). Anti-MT1-MMP monoclonal IgG{sub 3} and negative control IgG{sub 3} were radiolabelled with {sup 99m}Tc after derivatization with 6-hydrazinonicotinic acid (HYNIC) to yield {sup 99m}Tc-MT1-MMP mAb and {sup 99m}Tc-IgG{sub 3}, respectively. WHHLMI and control rabbits were injected with these radio-probes. The aorta was removed and radioactivity was measured at 24 h after the injection. Autoradiography and histological studies were performed. {sup 99m}Tc-MT1-MMP mAb accumulation in WHHLMI rabbit aortas was 5.4-fold higher than that of control rabbits. Regional {sup 99m}Tc-MT1-MMP mAb accumulation was positively correlated with MT1-MMP expression (r = 0.59, p < 0.0001), while {sup 99m}Tc-IgG{sub 3} accumulation was independent of MT1-MMP expression (r = 0.03, p = NS). The highest {sup 99m}Tc-MT1-MMP mAb accumulation was found in atheromatous lesions (4.8 {+-} 1.9, %ID x BW/mm{sup 2} x 10{sup 2}), followed in decreasing order by fibroatheromatous (1.8 {+-} 1.3), collagen-rich (1.6 {+-} 1.0) and neointimal lesions (1.5 {+-} 1.5). In contrast, {sup 99m}Tc-IgG{sub 3} accumulation was almost independent of the histological grade of lesions. Higher {sup 99m}Tc-MT1-MMP mAb accumulation in grade IV atheroma was shown in comparison with neointimal lesions or other more stable lesions. Nuclear imaging with {sup 99m}Tc-MT1-MMP mAb, in combination with CT and MRI, could provide new diagnostic imaging capabilities for detecting vulnerable plaques, although further investigations to improve target to blood ratios are strongly required. (orig.)

  3. Simvastatin induces NFκB/p65 down-regulation and JNK1/c-Jun/ATF-2 activation, leading to matrix metalloproteinase-9 (MMP-9) but not MMP-2 down-regulation in human leukemia cells.

    Science.gov (United States)

    Chen, Ying-Jung; Chang, Long-Sen

    2014-12-15

    The aim of the present study was to explore the signaling pathways associated with the effect of simvastatin on matrix metalloproteinase-2 (MMP-2)/MMP-9 expression in human leukemia K562 cells. In sharp contrast to its insignificant effect on MMP-2, simvastatin down-regulated MMP-9 protein expression and mRNA levels in K562 cells. Simvastatin-induced Pin1 down-regulation evoked NFκB/p65 degradation. Meanwhile, simvastatin induced JNK-mediated c-Jun and ATF-2 activation. Over-expression of Pin1 suppressed simvastatin-induced MMP-9 down-regulation. Treatment with SP600125 (a JNK inhibitor) or knock-down of JNK1 reduced MMP-2 expression in simvastatin-treated cells. Simvastatin enhanced the binding of c-Jun/ATF-2 with the MMP-2 promoter. Down-regulation of c-Jun or ATF-2 by siRNA revealed that c-Jun/ATF-2 activation was crucial for MMP-2 expression. Suppression of p65 activation or knock-down of Pin1 by shRNA reduced MMP-2 and MMP-9 expression in K562 cells. Over-expression of constitutively active JNK1 rescued MMP-2 expression in Pin1 shRNA-transfected cells. Simvastatin treatment also suppressed MMP-9 but not MMP-2 expression in human leukemia U937 and KU812 cells. Taken together, our data indicate that simvastatin-induced p65 instability leads to MMP-9 down-regulation in leukemia cells, while simvastatin-induced JNK1/c-Jun/ATF-2 activation maintains the MMP-2 expression underlying p65 down-regulation. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Effect of genistein on expression of MMP-9/TIMP-1 in rats with endometriosis%染料木黄酮对大鼠异位子宫内膜组织中MMP-9、TIMP-1表达的影响

    Institute of Scientific and Technical Information of China (English)

    郑兰; 朴松哲; 金延泽

    2012-01-01

    目的:探讨染料木黄酮(GEN)对患子宫内膜异位症(EMs)大鼠的基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶抑制剂-1(TIMP-1)表达水平的影响.方法:将68只Wistar大鼠分为正常组(A组,n=8)和EMs组(n=60),EMs组通过手术方法建立大鼠EMs模型,3周后测量移植物体积.EMs组大鼠随机分为4亚组:模型组(B组)和GEN低剂量组(C组,0.5mg/kg)、GEN中剂量组(D组,5mg/kg)、GEN高剂量组(E组,50mg/kg),连续皮下注射药物84d后处死大鼠,测量移植物体积、观察其组织学结构,并采用免疫组化法观察异位内膜组织中MMP -9、TIMP -1的表达.结果:与治疗前比较,E组移植物体积明显缩小(P<0.01),而C组和D组无差异;与A组比较,B组MMP -9表达率明显升高(P<0.05),TIMP-1表达率明显降低(P<0.01);与B组比较,E组MMP -9表达率明显降低,TIMP-1表达率明显升高(P<0.05),C组和D组无差异.结论:GEN可抑制大鼠EMs模型中异位内膜的生长,其抑制作用可能与MMP -9表达下降和TIMP -1表达升高,平衡MMP -9和TIMP -1的表达比例有关.%Objective: To explore the effect of genistein (GEN) on levels of matrix metallo proteinase -9 (MMP-9) and tissue inhibitor of metallo proteinase -1 (TIMP - 1) in rats with endomelriosis ( EMs). Methods: A total of 68 adult female Wistar rats were divided into the control group (group A, n =8) and EMs group (n =60), in which surgically induced EMs model was established. The rats in EMs group were divided into model group (group B), low dose GEN group ( group C, 0.5mg/kg), middle dose GEN group (group D, 5mg/kg) and high dose GEN group (group E, 50mg/kg). After daily administration of the corresponding agent for 84 days, the rats were sacrificed. The implant size of ectopic endometrium was measured and histological structure examination was conducted. The levels of MMP -9 and TIMP - 1 were evaluated with im-munohistochemistry. Results: After treatment, the implant size were significantly smaller in

  5. The regulation of matrix metalloproteinases by Id1 in gastric cancer%由Id1介导的MMP-2、MMP-9对胃癌生成的调控作用

    Institute of Scientific and Technical Information of China (English)

    雷婷; 韩霜; 郭雪艳; 丁杰

    2011-01-01

    Background and purpose: Angiogenesis is critical for the development of all malignancies.Id transcription factors have been shown to regulate key steps in tumor growth and metastasis due to their effects on angiogenesis.This study investigated the possible relationship between Id1 and matrix metalloproteinases (MMPs)in angiogenesis of gastric cancer.Methods: Expressions of Id1 and MMPs in gastric cancer tissues were examined by immunohistochemistry.The human gastric cancer SGC7901 sub-cell lines expressing Id1-siRNA was established.Expressions of MMP-2 and MMP-9 in Id1-siRNA transfectants were examined by semi-quantitative RT-PCR and Western blot.Enzyme activity of MMP-2 and MMP-9 in Id1-siRNA cells were detected by zymography method.Expressions of MMP-2 and MMP-9 in Id1-siRNA cell-transplanted tumor tissues in nude mice were identified by immunohistochemical method and Western blot.Results: Id1 and MMP-2, MMP-9 co-expressions were identified in the gastric cancer tissues.Significantly decreased expressions of both Id1 and MMP-2, MMP-9 on both protein and mRNA levels in SGC7901 Id1-siRNA transfectants were found.Data from zymography method showed that the activations of MMPs in Id1-siRNA cell lines were reduced compared to the control groups.Decreased expression of MMP-2 and MMP-9 in the Id1-siRNA transplanted tumor tissues in nude mice were found compared to those from the control cells.Conclusion: Id1 and MMP-2, MMP-9 were co-expressed in gastric cancer tissues; Id1 could upregulate the expression of MMP-2 and MMP-9 in gastric cancer.Downregulation of Id1 in the gastric cancer cells inhibited the transcriptional activities of MMP-2 and MMP-9.So we presumed that Id1 might promote tumorigenesis by transactivating MMP-2 and MMP-9 in gastric cancer.%背景与目的:肿瘤血管生成是肿瘤的一大重要特性,肿瘤微血管密度已被作为许多恶性肿瘤预后的一个重要指标.分化抑制因子(Id)是新近发现的与血管

  6. Presence of myofibroblasts and expression of matrix metalloproteinase-2 (MMP-2) in ameloblastomas correlate with rupture of the osseous cortical.

    Science.gov (United States)

    Fregnani, Eduardo Rodrigues; Sobral, Lays M; Alves, Fabio Abreu; Soares, Fernando Augusto; Kowalski, Luis Paulo; Coletta, Ricardo D

    2009-06-01

    Myofibroblasts are frequent in the stroma of neoplasm and by the expression of proteinases they can influence tumor infiltration and progression. In the present study, presence of myofibroblasts and expression of matrix metalloproteinase-2 (MMP-2) and urokinase plasminogen activator (uPA) were examined in intra-osseous solid multicystic ameloblastomas to determine their roles in the clinicopathological features of the tumors. Fifty seven ameloblastomas were analyzed immunohistochemically with antibodies against the isoform alpha of the smooth muscle actin (alpha-SMA), a specific marker of myofibroblasts, MMP-2 and uPA. Myofibroblasts were found in the stroma, in close contact with neoplastic cell islands, of approximately 58% (n = 33) of the ameloblastomas. MMP-2 and uPA were found in the cytoplasm of both neoplastic and stromal cells. A significant correlation between presence of myofibroblasts and MMP-2 expression was observed. Abundant presence of myofibroblast in the stroma of the tumors and expression of MMP-2 in the neoplastic or stromal cells were significantly correlated with rupture of the osseous cortical, which has been considered an important prognostic marker of ameloblastoma aggressiveness. Ours results suggest that abundant presence of myofibroblasts and expression of MMP-2 in solid ameloblastomas may be associated with a more aggressive infiltrative behavior.

  7. Clinical significance of serum levels of matrix metalloproteinase 2 (MMP-2) and its tissue inhibitor (TIMP-2) in gastric cancer.

    Science.gov (United States)

    Mroczko, Barbara; Lukaszewicz-Zając, Marta; Gryko, Mariusz; Kędra, Bogusław; Szmitkowski, Maciej

    2011-01-01

    Matrix metalloproteinase 2 (MMP-2) is able to degrade type IV collagen, and thus plays a key role in the migration of tumor cells. MMP-2 activity is inhibited by its tissue inhibitor (TIMP-2). The imbalance between MMPs and TIMPs may facilitate progression of cancer cells. The aim of this study was to compare the clinical importance of MMP-2 and TIMP-2 to that of classical tumor markers, namely carcinoembryonic antigen (CEA) and carbohydrate antigen (CA 19-9) in the diagnosis of gastric cancer (GC) by calculating the diagnostic criteria and estimating the levels of MMP-2, TIMP-2, CEA and CA 19-9 in GC patients in relation to clinicopathological features of cancer. We found that serum levels of MMP-2 and TIMP-2 were significantly lower, whereas serum tumor markers were higher, in GC patients than in healthy subjects. Moreover, concentrations of TIMP-2 and CEA correlated with gastric wall infiltration, while CA 19-9 levels correlated with gastric wall infiltration and the presence of nodal metastasis. None of the proteins tested was found to be an independent prognostic factor for GC patients' survival. The percentage of true positive results of TIMP-2 (61%) was higher than those of MMP-2 (54%) and the classical tumor markers CEA (21%) and CA 19-9 (31%). The highest diagnostic sensitivity was observed for the combined use of TIMP-2 with MMP-2 (77%). The results suggest the greater importance of serum MMP-2 and TIMP-2 than of the classical tumor markers CEA and CA 19-9 in the diagnosis of GC. But this issue requires further investigation.

  8. The possible role of matrix metalloproteinase (MMP)-2 and MMP-9 in cancer, e.g. acute leukemia

    NARCIS (Netherlands)

    Klein, G.; Vellenga, E.; Fraaije, M.W.; Kamps, W.A.; Bont, E.S.J.M. de

    2004-01-01

    In the past decades, a lot of effort has been put in identifying the role of matrix metalloproteinases (MMPs) in cancer. The main role of MMPs in angiogenesis, tumor growth and metastasis is degradation of extracellular matrix (ECM) and release and/or activation of growth factors through their

  9. Platelet-derived growth factor-D modulates extracellular matrix homeostasis and remodeling through TIMP-1 induction and attenuation of MMP-2 and MMP-9 gelatinase activities

    Energy Technology Data Exchange (ETDEWEB)

    Borkham-Kamphorst, Erawan, E-mail: ekamphorst@ukaachen.de; Alexi, Pascal; Tihaa, Lidia; Haas, Ute; Weiskirchen, Ralf, E-mail: rweiskirchen@ukaachen.de

    2015-02-13

    Platelet-derived growth factor-D (PDGF-D) is a more recent recognized growth factor involved in the regulation of several cellular processes, including cell proliferation, transformation, invasion, and angiogenesis by binding to and activating its cognate receptor PDGFR-β. After bile duct ligation or in the carbon tetrachloride-induced hepatic fibrosis model{sub ,} PDGF-D showed upregulation comparable to PDGF-B. Moreover, adenoviral PDGF-D gene transfer induced hepatic stellate cell proliferation and liver fibrosis. We here investigated the molecular mechanism of PDGF-D involvement in liver fibrogenesis. Therefore, the GRX mouse cell line was stimulated with PDGF-D and evaluated for fibrotic markers and PDGF-D signaling pathways in comparison to the other PDGF isoforms. We found that PDGF-D failed to enhance Col I and α-smooth muscle actin (α-SMA) production but has capacity to upregulate expression of the tissue inhibitor of metalloprotease 1 (TIMP-1) resulting in attenuation of MMP-2 and MMP-9 gelatinase activity as indicated by gelatinase zymography. This phenomenon was restored through application of a PDGF-D neutralizing antibody. Unexpectedly, PDGF-D incubation decreased both PDGFR-α and -β in mRNA and protein levels, and PDGF-D phosphorylated typrosines specific for PDGFR-α and -β. We conclude that PDGF-D intensifies fibrogenesis by interfering with the fibrolytic activity of the TIMP-1/MMP system and that PDGF-D signaling is mediated through both PDGF-α and -β receptors. - Highlights: • PDGF-D signals through PDGF receptor type α and β. • PDGF-D modulates extracellular matrix homeostasis and remodeling. • Like PDGF-B, PDGF-D triggers phosphorylation of PLC-γ, Akt/PKB, JNK, ERK1/2, and p38. • PDGF-D induces TIMP-1 expression through ERK and p38 MAPK. • PDGF-D attenuates MMP-2 and MMP-9 gelatinase activities.

  10. Association of Matrix Metalloproteinase-9 (MMP9 Variants with Primary Angle Closure and Primary Angle Closure Glaucoma.

    Directory of Open Access Journals (Sweden)

    Xueli Chen

    Full Text Available Shorter axial length observed in patients with primary angle closure glaucoma (PACG might be due to altered matrix metalloproteinase-9 (MMP9 activity resulting in ECM remodeling during eye growth and development. This study aimed to evaluate common variants in MMP9 for association with PACG. Six tag SNPs of MMP9 were genotyped in a Chinese sample of 1,030 cases, including 572 PACG and 458 primary angle closure (PAC, and 499 controls. None of 6 SNPs were significantly associated with overall PAC/PACG (P > 0.07 or with PAC/PACG subgroups (Pc > 0.18. Meta-analysis of two non-Chinese studies revealed significant association between rs17576 and PACG (ORs = 0.56, P 0.47. The largest association study to date did not find significant association between MMP9 and PAC/PACG in Chinese; meta-analysis with other Chinese datasets did not produce significant association. In most instances combination with non-Chinese datasets was not possible except for one variant showing nominally significant association. More work is needed to define the role of MMP9 variants in PACG.

  11. Data in support of the negative influence of divalent cations on (-)-epigallocatechin-3-gallate (EGCG)-mediated inhibition of matrix metalloproteinase-2 (MMP-2).

    Science.gov (United States)

    Deb, Gauri; Batra, Sahil; Limaye, Anil M

    2016-03-01

    In this data article we have provided evidence for the negative influence of divalent cations on (-)-epigallocatechin-3-gallate (EGCG)-mediated inhibition of matrix metalloproteinase-2 (MMP-2) activity in cell-free experiments. Chelating agents, such as EDTA and sodium citrate alone, did not affect MMP-2 activity. While EDTA enhanced, excess of divalent cations interfered with EGCG-mediated inhibition of MMP-2.

  12. Influence of Curcumin on Matrix Metalloproteinase(MMP)-2 and MMP-9 Expressions in Spinal Cord of Experimental Allergic Encephalomyelitis%姜黄素对EAE大鼠脊髓中MMP-2、MMP-9表达的影响

    Institute of Scientific and Technical Information of China (English)

    杨学志; 王赵伟; 李剑敏; 王贤亲; 张正学; 朱洁瑾

    2013-01-01

    Objective: To investigate the influence of curcumin on matrix metalloproteinase ( MMP ) -2 and MMP -9 expressions in spinal cord of experimental allergic encephalomyelitis (EAE) . Methods: The animal model was established in SD rats by injecting guinea pig spinal cord homogenate in complete Freund's adjuvant (CFA)and bordetella pertussis vaccine. Experimental group was given curcumin, and the changes of clinical symptoms were observed every day. Pathological changes of brain were observed by Hematoxylin and Eosin ( HE) staining. Real - time PCR was performed to test the transcriptional levels of MMP - 2 and MMP - 9 in cervical cord. Result: Compared with EAE group, the clinical scores and disease course were obviously reduced in curcumin group, furthermore the rats were recuperated quickly. The infiltration of inflammatory cells in central nerval system were obviously lessened. The transcriptional level of MMP - 9 was descend, the discrepancy of EAE and curcumins groups was significant, but the transcriptional level of MMP -2 in the two groups had no difference. Conclusion: Curcumin has therapeutic action on EAE, concerned with the inhibition of inflammatory cell infiltration and reducing the transcriptional level of MMP -9.%目的:探讨姜黄素对EAE大鼠脊髓中MMP-2、MMP-9活性的影响.方法:采用豚鼠脊髓匀浆诱导EAE模型,治疗组给予姜黄素进行干预,观察行为学变化,HE染色观察脑组织病理改变,real-time PCR检测颈髓组织MMP-2、MMP-9mRNA表达.结果:与EAE组相比,姜黄素治疗组临床评分明显下降,病程缩短,而且恢复较快;中枢炎性细胞浸润明显减少;MMP-9的转录水平明显下降,两组之间差异具有显著性,而MMP-2的水平两者无明显差异.结论:姜黄素对EAE具有一定的治疗作用,可能与抑制炎症细胞浸润及降低MMP-9的水平有关.

  13. The Inhibitory Effect of C-phycocyanin Containing Protein Extract (C-PC Extract) on Human Matrix Metalloproteinases (MMP-2 and MMP-9) in Hepatocellular Cancer Cell Line (HepG2).

    Science.gov (United States)

    Kunte, Mugdha; Desai, Krutika

    2017-03-30

    Spirulina platensis :have been studied for several biological activities. In the current study C-phycocyanin containing protein extract (C-PC extract) of Spirulina platensis have been studied for its effect on human matrix metalloproteinases (MMP-1, MMP-2 and MMP-9) and tissue inhibitors of MMPs (TIMP-1 and TIMP-2). In the present study, breast cancer cell line (MDA-MB 231) and hepatocellular cancer cell line (HepG2) were examined for inhibition of MMPs at different levels of expression after C-PC extract treatment. Herein, we have demonstrated that C-PC extract significantly reduced activity of MMP-2 by 55.13% and MMP-9 by 57.9% in HepG2 cells at 15 μg concentration. Additionally, the treatment has reduced mRNA expression of MMP-2 and MMP-9 at 20 μg concentration by 1.65-folds and 1.66-folds respectively. The C-PC extract treatment have also downregulated a mRNA expression of TIMP-2 by 1.12 folds at 20 μg concentration in HepG2 cells. Together, these results indicate that C-PC, extract successfully inhibited MMP-2 and -9 at different levels of expression and TIMP-2 at a mRNA expression level; however, extract did not have any effect on MMP-1 expressed in MDA-MB231 and TIMP-1 expressed in HepG2 cells as well as the exact mechanism of inhibition of MMP-2, MMP-9 and TIMP-2 remained unclear.

  14. Structural basis for matrix metalloproteinase-2 (MMP-2)-selective inhibitory action of β-amyloid precursor protein-derived inhibitor.

    Science.gov (United States)

    Hashimoto, Hiroshi; Takeuchi, Tomoka; Komatsu, Kyoko; Miyazaki, Kaoru; Sato, Mamoru; Higashi, Shouichi

    2011-09-23

    Unlike other synthetic or physiological inhibitors for matrix metalloproteinases (MMPs), the β-amyloid precursor protein-derived inhibitory peptide (APP-IP) having an ISYGNDALMP sequence has a high selectivity toward MMP-2. Our previous study identified amino acid residues of MMP-2 essential for its selective inhibition by APP-IP and demonstrated that the N to C direction of the decapeptide inhibitor relative to the substrate-binding cleft of MMP-2 is opposite that of substrate. However, detailed interactions between the two molecules remained to be clarified. Here, we determined the crystal structure of the catalytic domain of MMP-2 in complex with APP-IP. We found that APP-IP in the complex is indeed embedded into the substrate-binding cleft of the catalytic domain in the N to C direction opposite that of substrate. With the crystal structure, it was first clarified that the aromatic side chain of Tyr(3) of the inhibitor is accommodated into the S1' pocket of the protease, and the carboxylate group of Asp(6) of APP-IP coordinates bidentately to the catalytic zinc of the enzyme. The Ala(7) to Pro(10) and Tyr(3) to Ile(1) strands of the inhibitor extend into the nonprime and the prime sides of the cleft, respectively. Therefore, the decapeptide inhibitor has long range contact with the substrate-binding cleft of the protease. This mode of interaction is probably essential for the high MMP-2 selectivity of the inhibitor because MMPs share a common architecture in the vicinity of the catalytic center, but whole structures of their substrate-binding clefts have sufficient variety for the inhibitor to distinguish MMP-2 from other MMPs.

  15. Characterization of Porcine Matrix Metalloproteinase 23 (pMMP-23) Gene and Its Association with Litter Size Traits

    Institute of Scientific and Technical Information of China (English)

    NIU Bu-yue; LAN Xiao-ming; YAN Xiao-hong; DI Sheng-wei; WANG Yang; LI Hai-tao; XIONG Yuan-zhu; WANG Xi-biao

    2013-01-01

    The matrix metalloproteinase 23 (MMP-23), which might play a role in ovulation in mammals, was one of the promising candidate genes for litter size traits in pigs. In the present research, partial sequence of porcine MMP-23 (pMMP-23) gene, including exons 2-8 (GenBank:EU360790), was obtained. Real-time PCR analysis revealed that pMMP-23 gene was highly expressed in ovary. PCR-Sau3A I-RFLP and PCR-Acc II-RFLP assay were established to detect a C/T mutation in exon 3 (EU360790:g. 269C>T) and an A/G mutation in exon 4 (EU360790:g. 505A>G), respectively. Association study for these two SNPs with litter size was assessed in three independent populations (Minzhu, Landrace I and Landrace II). Statistical analysis demonstrated that for second and subsequent litters, TT sows produced more TNB than CC pigs in Landrace breed (PT locus, and the additive effect was significant (PG locus, and the additive effect was significant (PG locus might be a useful molecular marker for marker assisted selection (MAS).

  16. Global gene expression profiling of human pleural mesotheliomas: identification of matrix metalloproteinase 14 (MMP-14 as potential tumour target.

    Directory of Open Access Journals (Sweden)

    Stefania Crispi

    Full Text Available BACKGROUND: The goal of our study was to molecularly dissect mesothelioma tumour pathways by mean of microarray technologies in order to identify new tumour biomarkers that could be used as early diagnostic markers and possibly as specific molecular therapeutic targets. METHODOLOGY: We performed Affymetrix HGU133A plus 2.0 microarray analysis, containing probes for about 39,000 human transcripts, comparing 9 human pleural mesotheliomas with 4 normal pleural specimens. Stringent statistical feature selection detected a set of differentially expressed genes that have been further evaluated to identify potential biomarkers to be used in early diagnostics. Selected genes were confirmed by RT-PCR. As reported by other mesothelioma profiling studies, most of genes are involved in G2/M transition. Our list contains several genes previously described as prognostic classifier. Furthermore, we found novel genes, never associated before to mesotheliom that could be involved in tumour progression. Notable is the identification of MMP-14, a member of matrix metalloproteinase family. In a cohort of 70 mesothelioma patients, we found by a multivariate Cox regression analysis, that the only parameter influencing overall survival was expression of MMP14. The calculated relative risk of death in MM patients with low MMP14 expression was significantly lower than patients with high MMp14 expression (P = 0.002. CONCLUSIONS: Based on the results provided, this molecule could be viewed as a new and effective therapeutic target to test for the cure of mesothelioma.

  17. Global Gene Expression Profiling Of Human Pleural Mesotheliomas: Identification of Matrix Metalloproteinase 14 (MMP-14) as Potential Tumour Target

    Science.gov (United States)

    Crispi, Stefania; Calogero, Raffaele A.; Santini, Mario; Mellone, Pasquale; Vincenzi, Bruno; Citro, Gennaro; Vicidomini, Giovanni; Fasano, Silvia; Meccariello, Rosaria; Cobellis, Gilda; Menegozzo, Simona; Pierantoni, Riccardo; Facciolo, Francesco; Baldi, Alfonso; Menegozzo, Massimo

    2009-01-01

    Background The goal of our study was to molecularly dissect mesothelioma tumour pathways by mean of microarray technologies in order to identify new tumour biomarkers that could be used as early diagnostic markers and possibly as specific molecular therapeutic targets. Methodology We performed Affymetrix HGU133A plus 2.0 microarray analysis, containing probes for about 39,000 human transcripts, comparing 9 human pleural mesotheliomas with 4 normal pleural specimens. Stringent statistical feature selection detected a set of differentially expressed genes that have been further evaluated to identify potential biomarkers to be used in early diagnostics. Selected genes were confirmed by RT-PCR. As reported by other mesothelioma profiling studies, most of genes are involved in G2/M transition. Our list contains several genes previously described as prognostic classifier. Furthermore, we found novel genes, never associated before to mesotheliom that could be involved in tumour progression. Notable is the identification of MMP-14, a member of matrix metalloproteinase family. In a cohort of 70 mesothelioma patients, we found by a multivariate Cox regression analysis, that the only parameter influencing overall survival was expression of MMP14. The calculated relative risk of death in MM patients with low MMP14 expression was significantly lower than patients with high MMp14 expression (P = 0.002). Conclusions Based on the results provided, this molecule could be viewed as a new and effective therapeutic target to test for the cure of mesothelioma. PMID:19753302

  18. Distal Interleukin-1β (IL-1β) Response Element of Human Matrix Metalloproteinase-13 (MMP-13) Binds Activator Protein 1 (AP-1) Transcription Factors and Regulates Gene Expression*

    Science.gov (United States)

    Schmucker, Adam C.; Wright, Jason B.; Cole, Michael D.; Brinckerhoff, Constance E.

    2012-01-01

    The collagenase matrix metalloproteinase-13 (MMP-13) plays an important role in the destruction of cartilage in arthritic joints. MMP-13 expression is strongly up-regulated in arthritis, largely because of stimulation by inflammatory cytokines such as IL-1β. Treatment of chondrocytes with IL-1β induces transcription of MMP-13 in vitro. IL-1β signaling converges upon the activator protein-1 transcription factors, which have been shown to be required for IL-1β-induced MMP-13 gene expression. Using chromatin immunoprecipitation (ChIP), we detected activator protein-1 binding within an evolutionarily conserved DNA sequence ∼20 kb 5′ relative to the MMP-13 transcription start site (TSS). Also using ChIP, we detected histone modifications and binding of RNA polymerase II within this conserved region, all of which are consistent with transcriptional activation. Chromosome conformation capture indicates that chromosome looping brings this region in close proximity with the MMP-13 TSS. Finally, a luciferase reporter construct driven by a component of the conserved region demonstrated an expression pattern similar to that of endogenous MMP-13. These data suggest that a conserved region at 20 kb upstream from the MMP-13 TSS includes a distal transcriptional response element of MMP-13, which contributes to MMP-13 gene expression. PMID:22102411

  19. The effect of stromelysin-1 (MMP-3) on non-collagenous extracellular matrix proteins of demineralized dentin and the adhesive properties of restorative resins.

    Science.gov (United States)

    Boukpessi, T; Menashi, S; Camoin, L; Tencate, J M; Goldberg, M; Chaussain-Miller, C

    2008-11-01

    Dentin non-collagenous matrix components (NCPs) are structural proteins involved in the formation, the architecture and the mineralization of the extracellular matrix (ECM). We investigated here how recombinant metalloproteinase stromelysin-1, also termed MMP-3, initiates the release of ECM molecules from artificially demineralized human dentin. Analysis of the supernatants by Western blotting reveals that MMP-3 extracts PGs (decorin, biglycan), and also a series of phosphorylated proteins: dentin sialoprotein (DSP), osteopontin (OPN), bone sialoprotein (BSP) and MEPE, but neither dentin matrix protein-1 (DMP1), another member of the SIBLING family, nor osteocalcin (OC), a non-phosphorylated matrix molecule. After treatment of dentin surfaces by MMP-3, scanning electron microscope (SEM) examination of resin replica shows an increased penetration of the resin into the dentin tubules when compared to surfaces only treated by demineralizing solutions. This preclinical investigation suggests that MMP-3 may be used to improve the adhesive properties of restorative materials.

  20. Serum levels and tissue expression of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinases 2 (TIMP-2) in colorectal cancer patients.

    Science.gov (United States)

    Groblewska, Magdalena; Mroczko, Barbara; Gryko, Mariusz; Pryczynicz, Anna; Guzińska-Ustymowicz, Katarzyna; Kędra, Bogusław; Kemona, Andrzej; Szmitkowski, Maciej

    2014-04-01

    The objective of the study was the assessment of serum levels and tissue expression of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of matrix metalloproteinases 2 (TIMP-2) in patients with colorectal cancer (CRC). The study included 72 CRC patients and 68 healthy subjects. The serum levels of MMP-2 and TIMP-2 were measured using enzyme-linked immunosorbent assay (ELISA) method, whereas tissue expression of MMP-2 and TIMP-2 in cancer cells, interstitial inflammatory cells, and adjacent normal colorectal mucosa were examined by immunohistochemical staining of tumor samples. The serum levels of MMP-2 and TIMP-2 in cancer patients were significantly lower than those in control group, but the percentage of positive immunoreactivity of these proteins were higher in malignant and inflammatory cells as compared to normal tissue. There was a significant correlation between MMP-2 immunoreactivity in inflammatory cells and the presence of distant metastases and between TIMP-2 expression in inflammatory cells and tumor size, nodal involvement, and distant metastases. Area under receiver operating characteristic (ROC) curve (AUC) for serum MMP-2 was higher than for serum TIMP-2. Moreover, positive tissue expression of MMP-2 was a significant prognostic factor for CRC patients' survival. Our findings suggest that MMP-2 and TIMP-2 might play a role in the process of colorectal cancer invasion and metastasis, but the significance of their interactions with tumor stroma and interstitial inflammatory infiltration in colorectal neoplasia require further elucidation.

  1. Matrix metalloproteinase-10 (MMP-10) interaction with tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2: binding studies and crystal structure.

    Science.gov (United States)

    Batra, Jyotica; Robinson, Jessica; Soares, Alexei S; Fields, Alan P; Radisky, Derek C; Radisky, Evette S

    2012-05-04

    Matrix metalloproteinase 10 (MMP-10, stromelysin-2) is a secreted metalloproteinase with functions in skeletal development, wound healing, and vascular remodeling; its overexpression is also implicated in lung tumorigenesis and tumor progression. To understand the regulation of MMP-10 by tissue inhibitors of metalloproteinases (TIMPs), we have assessed equilibrium inhibition constants (K(i)) of putative physiological inhibitors TIMP-1 and TIMP-2 for the active catalytic domain of human MMP-10 (MMP-10cd) using multiple kinetic approaches. We find that TIMP-1 inhibits the MMP-10cd with a K(i) of 1.1 × 10(-9) M; this interaction is 10-fold weaker than the inhibition of the similar MMP-3 (stromelysin-1) catalytic domain (MMP-3cd) by TIMP-1. TIMP-2 inhibits the MMP-10cd with a K(i) of 5.8 × 10(-9) M, which is again 10-fold weaker than the inhibition of MMP-3cd by this inhibitor (K(i) = 5.5 × 10(-10) M). We solved the x-ray crystal structure of TIMP-1 bound to the MMP-10cd at 1.9 Å resolution; the structure was solved by molecular replacement and refined with an R-factor of 0.215 (R(free) = 0.266). Comparing our structure of MMP-10cd·TIMP-1 with the previously solved structure of MMP-3cd·TIMP-1 (Protein Data Bank entry 1UEA), we see substantial differences at the binding interface that provide insight into the differential binding of stromelysin family members to TIMP-1. This structural information may ultimately assist in the design of more selective TIMP-based inhibitors tailored for specificity toward individual members of the stromelysin family, with potential therapeutic applications.

  2. Identification and characterization of a matrix metalloproteinase (Pta1-MMP) expressed during Loblolly pine (Pinus taeda) seed development, germination completion, and early seedling establishment.

    Science.gov (United States)

    Ratnaparkhe, Supriya M; Egertsdotter, E M Ulrika; Flinn, Barry S

    2009-07-01

    Extracellular matrix (ECM) modifications occur during plant growth, development, and in response to environmental stimuli. Key modulators of ECM modification in vertebrates, the extracellular matrix metalloproteinases (MMPs), have also been described in a few plants. Here, we report the identification of Loblolly pine (Pinus taeda) Pta1-MMP and its characterization during seed development and germination. Pta1-MMP protein has the structural characteristics of other plant MMPs, the recombinant protein exhibits Zn(2+)-dependent protease activity, and is inhibited by EDTA and the active site-binding hydroxamate inhibitor GM6001. The Pta1-MMP gene is expressed in both embryo and megagametophyte, with transcript levels increasing in both during the period from proembryo to early cotyledonary stage, then declining during late embryogenesis and maturation drying. Protein extracts exhibited similar developmental-stage MMP-like activity. Seed germination was stimulated by GA(3) and inhibited by ABA, and the timing of germination completion was mirrored by the presence of MMP-like protease activity in both water- and GA(3)-imbibed embryos. Pta1-MMP gene transcript levels increased in association with radicle protrusion for both GA(3)- and water-treated embryos, in agreement with MMP-like activity. In contrast, by 11 days after imbibition, Pta1-MMP gene transcripts in ABA-treated embryos were at levels similar to the other treatments, although MMP-like activity was not observed. The application of GM6001 during Loblolly pine seed germination inhibited radicle protrusion. Our results suggest that MMP activity may be involved in ECM modification, facilitating the cell division and expansion required during seed development, germination completion, and subsequent seedling establishment.

  3. Expression of Matrix Metalloproteinase 1(MMP1 in HepatoCellular Carcinoma (HCC: Immunohistochemical and Biochemical Studies

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    Mohy Eldin abdel Fattah abdel Atty Yassen*, Olfat Ali Ibrahim Hammam** and Hazem Kamel Abdel-Aziz Mohamed Sarhan***.

    2013-04-01

    Full Text Available Background: The present study aimed to evaluate the expression of Matrix Metalloproteinase-1 (MMP1 in HepatoCellular Carcinoma (HCC by using the Immunohistochemical technique, which allows us to integrate the biological aspects of this enzymatic expression in the morphological context of HCCs.Material and Methods: The study was performed on 70 subjects from out and in patients of Tropical medicine Department, Thiodor Billhars Institute during the period from January 2011 until June 2012. The present study included 60 patients with chronic hepatitis C who had undergone liver biopsy. They consisted of 42 men and 28 women with ages ranging from 36 to 66 years. The diagnosis of chronic hepatitis C was made on the basis of positivity for anti-HCV (by the second generation ELISA, and confirmed by HCV-RNA reverse transcription polymerase chain reaction (RT-PCR. Patients were divided into four groups: Group I: included 10 normal persons with no history of liver disease with normal liver enzymes and free ultrasonographic finding as normal control. It included 6 males, 4 females, with ages ranging from 34 to 48 years. Group II: included 20 HCV infected patients without cirrhotic changes. It included 11 males, 9 females, with ages ranging from 39 to 53 years. Group III: included 20 HCV infected patients with liver cirrhosis, 12 males, 8 females, with ages ranging from 48-63 years. Group IV: included 20 HCV infected patients with HCC, 16 males, 4 females, with ages ranging from 53-64 years.Results: Blood Picture, (Hb, WBCs, RBCs, Plts, PC and ESR. Liver Function Test (ALT, AST, ALB, GGT, ALP, T. BIL and D. BIL. Matrix Metalloprotenase 1(MMP1 Measurements: Serum MMP1. Histopathological investigation Including histopathological changes in the liver tissue.Conclusion: our results suggest that MMP-1 is overexpressed in a large proportion of patients with HCC and the high expression level of protein correlated with the disease progression and poor clinical

  4. Downregulation of matrix metalloproteinase-2 (MMP-2) utilizing adenovirus-mediated transfer of small interfering RNA (siRNA) in a novel spinal metastatic melanoma model.

    Science.gov (United States)

    Tsung, Andrew J; Kargiotis, Odysseas; Chetty, Chandramu; Lakka, Sajani S; Gujrati, Meena; Spomar, Daniel G; Dinh, Dzung H; Rao, Jasti S

    2008-03-01

    Matrix metalloproteinases (MMPs) comprise a class of secreted zinc-dependent endopeptidases implicated in the metastatic potential of tumor cells due to their ability to degrade the extracellular matrix (ECM) and basement membrane. Matrix metalloproteinase-2 (MMP-2) has been detected in high levels and correlates with invasiveness in human melanoma. We have studied the effect of adenovirus-mediated transfer of small interfering RNA (siRNA) against MMP-2 in the human melanoma cell line A2058. The delivery of these double-stranded RNA molecules represents an efficient technology in silencing disease-causing genes with known sequences at the post-transcriptional level. siRNA against MMP-2 mRNA (Ad-MMP-2) was found to decrease MMP-2 protein expression and activity in melanoma cells as demonstrated by western blotting and gelatin zymography. Furthermore, infection of cells with Ad-MMP-2 inhibited cellular migration and invasion as indicated by spheroid and matrigel assays. We also observed dose-dependent suppression of vascular network formation in an angiogenesis assay. Finally, we developed a nude mouse spinal metastatic model to investigate the local effects of tumor metastasis. Intravenous tail vein injection with Ad-MMP-2 on days 5, 9 and 11 after tumor implantation resulted in complete retention of neurological function as compared to control and scrambled vector (Ad-SV)-treated groups that showed complete paraplegia by day 14+/-2 days. Hematoxylin and eosin staining revealed decreased tumor size in the Ad-MMP-2-treated animals. This novel experimental model revealed that adenoviral-mediated transfer of RNA interference against MMP-2 results in the retention of neurological function and significantly inhibited tumor growth.

  5. TNF-{alpha} promotes human retinal pigment epithelial (RPE) cell migration by inducing matrix metallopeptidase 9 (MMP-9) expression through activation of Akt/mTORC1 signaling

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Cheng-hu; Cao, Guo-Fan [The Affiliated Eye Hospital of Nanjing Medical University, Nanjing 210029 (China); Jiang, Qin, E-mail: Jqin710@vip.sina.com [The Affiliated Eye Hospital of Nanjing Medical University, Nanjing 210029 (China); Yao, Jin, E-mail: dryaojin@yahoo.com [The Affiliated Eye Hospital of Nanjing Medical University, Nanjing 210029 (China)

    2012-08-17

    Highlights: Black-Right-Pointing-Pointer TNF-{alpha} induces MMP-9 expression and secretion to promote RPE cell migration. Black-Right-Pointing-Pointer MAPK activation is not critical for TNF-{alpha}-induced MMP-9 expression. Black-Right-Pointing-Pointer Akt and mTORC1 signaling mediate TNF-{alpha}-induced MMP-9 expression. Black-Right-Pointing-Pointer SIN1 knockdown showed no significant effect on MMP-9 expression by TNF-{alpha}. -- Abstract: Tumor necrosis factor-alpha (TNF-{alpha}) promotes in vitro retinal pigment epithelial (RPE) cell migration to initiate proliferative vitreoretinopathy (PVR). Here we report that TNF-{alpha} promotes human RPE cell migration by inducing matrix metallopeptidase 9 (MMP-9) expression. Inhibition of MMP-9 by its inhibitor or its neutralizing antibody inhibited TNF-{alpha}-induced in vitro RPE cell migration. Reversely, exogenously-added active MMP-9 promoted RPE cell migration. Suppression Akt/mTOR complex 1(mTORC1) activation by LY 294002 and rapamycin inhibited TNF-{alpha}-mediated MMP-9 expression. To introduce a constitutively active Akt (CA-Akt) in cultured RPE cells increased MMP-9 expression, and to block mTORC1 activation by rapamycin inhibited its effect. RNA interference (RNAi)-mediated silencing of SIN1, a key component of mTOR complex 2 (mTORC2), had no effect on MMP-9 expression or secretion. In conclusion, this study suggest that TNF-{alpha} promotes RPE cell migration by inducing MMP-9 expression through activation of Akt/ mTORC1, but not mTORC2 signaling.

  6. Genetic variants associated with circulating MMP1 levels near matrix metalloproteinase genes on chromosome 11q21-22 in Taiwanese: interaction with obesity

    Science.gov (United States)

    2013-01-01

    Background MMP1 is implicated in the pathogenesis of atherothrombotic cardiovascular disease. We aimed to elucidate genetic determinants of inflammatory marker levels, including circulating MMP1, in Taiwanese, and their association with obesity. Methods Five genetic polymorphisms around matrix metalloproteinase genes on chromosome 11q21-22 region were genotyped in 519 subjects. Results After adjusting for clinical covariates, two polymorphisms were significantly associated with MMP1 levels, rs1799750 and rs495366, using an additive inheritance model (P = 1.5x10-4 and P = 2.57x10-5, respectively). Using dominant model, minor alleles of rs1799750 and rs495366 were associated with higher MMP1 levels (P = 1.3x10-4 and P = 1.95x10-5, respectively). In haplotype analysis, two haplotypes inferred from five SNPs (A2GATA and A1GATG) were associated with MMP1 levels (P = 5x10-4 and P = 8.47x10-5, respectively). Subgroup and interaction analysis revealed an association of rs1799750 and rs495366 with MMP1 levels only in non-obese subjects (P = 6.66x10-6 and P = 4.38x10-5, respectively, and interaction P = 0.008 for rs1799750). Haplotype interaction analysis also showed significant interaction for haplotype A1GATG (interaction P = 0.003). Conclusions Genotypes/haplotypes around MMP1 locus are associated with MMP1 levels in Taiwanese. Further, since genotypes/haplotypes near MMP1 locus interact with obesity to set MMP1 levels, genetic determinants for MMP1 level may be different between obese and non-obese individuals. PMID:23497408

  7. Matrix metalloproteinase 2 (MMP-2) degrades soluble vasculotropic amyloid-beta E22Q and L34V mutants, delaying their toxicity for human brain microvascular endothelial cells.

    Science.gov (United States)

    Hernandez-Guillamon, Mar; Mawhirt, Stephanie; Fossati, Silvia; Blais, Steven; Pares, Mireia; Penalba, Anna; Boada, Merce; Couraud, Pierre-Olivier; Neubert, Thomas A; Montaner, Joan; Ghiso, Jorge; Rostagno, Agueda

    2010-08-27

    Patients carrying mutations within the amyloid-beta (Abeta) sequence develop severe early-onset cerebral amyloid angiopathy with some of the related variants manifesting primarily with hemorrhagic phenotypes. Matrix metalloproteases (MMPs) are typically associated with blood brain barrier disruption and hemorrhagic transformations after ischemic stroke. However, their contribution to cerebral amyloid angiopathy-related hemorrhage remains unclear. Human brain endothelial cells challenged with Abeta synthetic homologues containing mutations known to be associated in vivo with hemorrhagic manifestations (AbetaE22Q and AbetaL34V) showed enhanced production and activation of MMP-2, evaluated via Multiplex MMP antibody arrays, gel zymography, and Western blot, which in turn proteolytically cleaved in situ the Abeta peptides. Immunoprecipitation followed by mass spectrometry analysis highlighted the generation of specific C-terminal proteolytic fragments, in particular the accumulation of Abeta-(1-16), a result validated in vitro with recombinant MMP-2 and quantitatively evaluated using deuterium-labeled internal standards. Silencing MMP-2 gene expression resulted in reduced Abeta degradation and enhanced apoptosis. Secretion and activation of MMP-2 as well as susceptibility of the Abeta peptides to MMP-2 degradation were dependent on the peptide conformation, with fibrillar elements of AbetaE22Q exhibiting negligible effects. Our results indicate that MMP-2 release and activation differentially degrades Abeta species, delaying their toxicity for endothelial cells. However, taking into consideration MMP ability to degrade basement membrane components, these protective effects might also undesirably compromise blood brain barrier integrity and precipitate a hemorrhagic phenotype.

  8. The effect of stromelysin-1 (MMP-3) on non-collagenous extracellular matrix proteins of demineralized dentin and the adhesive properties of restorative resins

    NARCIS (Netherlands)

    T. Boukpessi; S. Menashi; L. Camoin; J.M. ten Cate; M. Goldberg; C. Chaussain-Miller

    2008-01-01

    Dentin non-collagenous matrix components (NCPs) are structural proteins involved in the formation, the architecture and the mineralization of the extracellular matrix (ECM). We investigated here how recombinant metalloproteinase stromelysin-1, also termed MMP-3, initiates the release of ECM molecule

  9. The significance of determination of Matrix Metalloproteinase-3(MMP-3)content in lumbar disc herniation.%腰椎间盘突出症血清MMP-3测定的意义

    Institute of Scientific and Technical Information of China (English)

    郑昱新; 石印玉

    2001-01-01

    Objective To investigate the relationship between sero-matrix mefalloproteinase-3(MMP-3) content and lumbar disc herniation.Method The sero-MMP-3 content in 32 patients with lumbar disc herniation and 10 normal human were determined with enzyme-immunoassay method.Result The mean sero-MMP-3 content in lumbar disc herniation was 35.57ng/ml,while normal human27.56ng/ml,the deference was siginificant (P<0.01).However the sero-MMP-3 content in lumbar disc prolapsus was siginificantly higher than that in nonprolapsus(P<0.05).Conclusion The determination of sero-MMP-3 content can be used as assistant diagnosis of the lumbar disc herniation,furthermore confirm the relationship between sero-MMP-3 content and degree of lumbar disc degeneration.%目的 进一步研究血清MMP-3的含量与腰椎间盘突出症发病的关系。方法 32例腰椎间盘突出症患者和10例正常人,用酶免疫法测定他们血清MMP-3的含量。结果 腰椎间盘突出症患者血清MMP-3含量的平均值为:35.57ng/ml,而正常人平均值为:27.56ng/ml,两者的差异有显著性意义(P<0.01)。其中脱出型腰椎间盘突出症患者的血清MMP-3含量明显高于非脱出型患者(P<0.05)。结论 血清MMP-3的测定可作为对腰椎间盘突出症的辅助诊断,并且证实血清MMP-3的含量与腰椎间盘的退变程度相关。

  10. Comparative Analysis of Matrix Metalloproteinase Family Members Reveals That MMP9 Predicts Survival and Response to Temozolomide in Patients with Primary Glioblastoma

    Science.gov (United States)

    Cai, Jinquan; Sun, Ying; Wang, Guangzhi; Li, Yongli; Li, Ruiyan; Feng, Yan; Han, Bo; Li, Jianlong; Tian, Yu; Yi, Liye; Jiang, Chuanlu

    2016-01-01

    Background Glioblastoma multiform (GBM) is the most common malignant primary brain tumor in adults. Radiotherapy plus concomitant and adjuvant TMZ chemotherapy is the current standard of care for patients with GBM. Matrix metalloproteinases (MMPs), a family of zinc-dependent endopeptidases, are key modulators of tumor invasion and metastasis due to their ECM degradation capacity. The aim of the present study was to identify the most informative MMP member in terms of prognostic and predictive ability for patients with primary GBM. Method The mRNA expression profiles of all MMP genes were obtained from the Chinese Glioma Genome Atlas (CGGA), the Repository for Molecular Brain Neoplasia Data (REMBRANDT) and the GSE16011 dataset. MGMT methylation status was also examined by pyrosequencing. The correlation of MMP9 expression with tumor progression was explored in glioma specimens of all grades. Kaplan–Meier analysis and Cox proportional hazards regression models were used to investigate the association of MMP9 expression with survival and response to temozolomide. Results MMP9 was the only significant prognostic factor in three datasets for primary glioblastoma patients. Our results indicated that MMP9 expression is correlated with glioma grade (p<0.0001). Additionally, low expression of MMP9 was correlated with better survival outcome (OS: p = 0.0012 and PFS: p = 0.0066), and MMP9 was an independent prognostic factor in primary GBM (OS: p = 0.027 and PFS: p = 0.032). Additionally, the GBM patients with low MMP9 expression benefited from temozolomide (TMZ) chemotherapy regardless of the MGMT methylation status. Conclusions Patients with primary GBMs with low MMP9 expression may have longer survival and may benefit from temozolomide chemotherapy. PMID:27022952

  11. Matrix metalloproteinase-10/TIMP-2 structure and analyses define conserved core interactions and diverse exosite interactions in MMP/TIMP complexes.

    Science.gov (United States)

    Batra, Jyotica; Soares, Alexei S; Mehner, Christine; Radisky, Evette S

    2013-01-01

    Matrix metalloproteinases (MMPs) play central roles in vertebrate tissue development, remodeling, and repair. The endogenous tissue inhibitors of metalloproteinases (TIMPs) regulate proteolytic activity by binding tightly to the MMP active site. While each of the four TIMPs can inhibit most MMPs, binding data reveal tremendous heterogeneity in affinities of different TIMP/MMP pairs, and the structural features that differentiate stronger from weaker complexes are poorly understood. Here we report the crystal structure of the comparatively weakly bound human MMP-10/TIMP-2 complex at 2.1 Å resolution. Comparison with previously reported structures of MMP-3/TIMP-1, MT1-MMP/TIMP-2, MMP-13/TIMP-2, and MMP-10/TIMP-1 complexes offers insights into the structural basis of binding selectivity. Our analyses identify a group of highly conserved contacts at the heart of MMP/TIMP complexes that define the conserved mechanism of inhibition, as well as a second category of diverse adventitious contacts at the periphery of the interfaces. The AB loop of the TIMP N-terminal domain and the contact loops of the TIMP C-terminal domain form highly variable peripheral contacts that can be considered as separate exosite interactions. In some complexes these exosite contacts are extensive, while in other complexes the AB loop or C-terminal domain contacts are greatly reduced and appear to contribute little to complex stability. Our data suggest that exosite interactions can enhance MMP/TIMP binding, although in the relatively weakly bound MMP-10/TIMP-2 complex they are not well optimized to do so. Formation of highly variable exosite interactions may provide a general mechanism by which TIMPs are fine-tuned for distinct regulatory roles in biology.

  12. [Matrix metalloproteinases (MMP)--MMP-1,-2,-9 and its endogenous activity regulators in transformed by E7 oncogene HPV16 and HPV18 cervical carcinoma cell lines].

    Science.gov (United States)

    Ryzhakova, O S; Solov'eva, N I

    2013-01-01

    Matrix metalloproteinases (MMP) play a key role in development of tumor invasion and metestasis. The purpose of the work is the elucidation of peculiarities of expression of MMP-1, MMP-2, MMP-9 and their activity regulators: plasminogen activator uPA and tissue inhibitors of MMPs - TIMP-1 and TIMP-2 in human cell lines of squoamous cell carcinoma (SCC). Comparative study of MMPs' expression was carried out on cell lines SCC which differed in HPV types (HPV-16 and HPV-18): SiHa, Caski - HPV16, Hela, C4-1 - HPV18). As a control, the C33A line was used where HPV copies were absent. The human papilloma viruses (HPV) of high risk--HPV-16, HPV-18, as etiological factors of initiation of cervical cancer, are most widespread and most aggressive among oncogenic HPVs. Study of MMP expression involved estimation of expression of mRNA using the RT-PCR method and determination of collagenolytic activity by hydrolysis of fluorogenic type 1 collagen and also by the zymography method. It was shown that: 1. In both types of cell lines, the MMP-1 expression was essentially increased (2 to 8 times), and in HPV18 lines it was most expressed. The exception was made by the SiHa line in which the decrease of expression of this enzyme was observed. MMP-2 expression was at the control level in both types of cell lines. 2. Expression of inhibitors generally was at the control level. The only exception was the C4-1 line where the expression of TIMP-1 and TIMP-2 was increased in 1,7 and 2,6 times accordingly. Expression of uPA was increased 2 to 4, 5 times in all cell lines except Siha where was lowered to 20%. 3. Collagenolytic activity in the Caski and Hela cell line was 2-3 times higher that it was in control, while the activity in the SiHa cell line was compatible with that in the control. Research of gelatinolytic activity also as well as the data on an expression MPHK has revealed only presence MMFP-2, but not MMP-9 in all cervical carcinoma cell lines. The data obtained provide

  13. Plasma levels of mature brain-derived neurotrophic factor (BDNF) and matrix metalloproteinase-9 (MMP-9) in treatment-resistant schizophrenia treated with clozapine.

    Science.gov (United States)

    Yamamori, Hidenaga; Hashimoto, Ryota; Ishima, Tamaki; Kishi, Fukuko; Yasuda, Yuka; Ohi, Kazutaka; Fujimoto, Michiko; Umeda-Yano, Satomi; Ito, Akira; Hashimoto, Kenji; Takeda, Masatoshi

    2013-11-27

    Brain-derived neurotrophic factor (BDNF) regulates the survival and growth of neurons, and influences synaptic efficiency and plasticity. Peripheral BDNF levels in patients with schizophrenia have been widely reported in the literature. However, it is still controversial whether peripheral levels of BDNF are altered in patients with schizophrenia. The peripheral BDNF levels previously reported in patients with schizophrenia were total BDNF (proBDNF and mature BDNF) as it was unable to specifically measure mature BDNF due to limited BDNF antibody specificity. In this study, we examined whether peripheral levels of mature BDNF were altered in patients with treatment-resistant schizophrenia. Matrix metalloproteinase-9 (MMP-9) levels were also measured, as MMP-9 plays a role in the conversion of proBDNF to mature BDNF. Twenty-two patients with treatment-resistant schizophrenia treated with clozapine and 22 age- and sex-matched healthy controls were enrolled. The plasma levels of mature BDNF and MMP-9 were measured using ELISA kits. No significant difference was observed for mature BDNF however, MMP-9 was significantly increased in patients with schizophrenia. The significant correlation was observed between mature BDNF and MMP-9 plasma levels. Neither mature BDNF nor MMP-9 plasma levels were associated clinical variables. Our results do not support the view that peripheral BDNF levels are associated with schizophrenia. MMP-9 may play a role in the pathophysiology of schizophrenia and serve as a biomarker for schizophrenia.

  14. A cellular Potts model for the MMP-dependent and -independent cancer cell migration in matrix microtracks of different dimensions

    Science.gov (United States)

    Scianna, Marco; Preziosi, Luigi

    2014-03-01

    Cell migration is fundamental in a wide variety of physiological and pathological phenomena, among other in cancer invasion and development. In particular, the migratory/invasive capability of single metastatic cells is fundamental in determining the malignancy of a solid tumor. Specific cell migration phenotypes result for instance from the reciprocal interplay between the biophysical and biochemical properties of both the malignant cells themselves and of the surrounding environment. In particular, the extracellular matrices (ECMs) forming connective tissues can provide both loosely organized zones and densely packed barriers, which may impact cell invasion mode and efficiency. The critical processes involved in cell movement within confined spaces are (i) the proteolytic activity of matrix metalloproteinases (MMPs) and (ii) the deformation of the entire cell body, and in particular of the nucleus. We here present an extended cellular Potts model (CPM) to simulate a bio-engineered matrix system, which tests the active motile behavior of a single cancer cell into narrow channels of different widths. As distinct features of our approach, the cell is modeled as a compartmentalized discrete element, differentiated in the nucleus and in the cytosolic region, while a directional shape-dependent movement is explicitly driven by the evolution of its polarity vector. As outcomes, we find that, in a large track, the tumor cell is not able to maintain a directional movement. On the contrary, a structure of subcellular width behaves as a contact guidance sustaining cell persistent locomotion. In particular, a MMP-deprived cell is able to repolarize and follow the micropattern geometry, while a full MMP activity leads to a secondary track expansion by degrading the matrix structure. Finally, we confirm that cell movement within a subnuclear structure can be achieved either by pericellular proteolysis or by a significant deformation of cell nucleus.

  15. MMP-sensitive PEG diacrylate hydrogels with spatial variations in matrix properties stimulate directional vascular sprout formation.

    Directory of Open Access Journals (Sweden)

    Michael V Turturro

    Full Text Available The spatial presentation of immobilized extracellular matrix (ECM cues and matrix mechanical properties play an important role in directed and guided cell behavior and neovascularization. The goal of this work was to explore whether gradients of elastic modulus, immobilized matrix metalloproteinase (MMP-sensitivity, and YRGDS cell adhesion ligands are capable of directing 3D vascular sprout formation in tissue engineered scaffolds. PEGDA hydrogels were engineered with mechanical and biofunctional gradients using perfusion-based frontal photopolymerization (PBFP. Bulk photopolymerized hydrogels with uniform mechanical properties, degradation, and immobilized biofunctionality served as controls. Gradient hydrogels exhibited an 80.4% decrease in elastic modulus and a 56.2% decrease in immobilized YRGDS. PBFP hydrogels also demonstrated gradients in hydrogel degradation with degradation times ranging from 10-12 hours in the more crosslinked regions to 4-6 hours in less crosslinked regions. An in vitro model of neovascularization, composed of co-culture aggregates of endothelial and smooth muscle cells, was used to evaluate the effect of these gradients on vascular sprout formation. Aggregate invasion in gradient hydrogels occurred bi-directionally with sprout alignment observed in the direction parallel to the gradient while control hydrogels with homogeneous properties resulted in uniform invasion. In PBFP gradient hydrogels, aggregate sprout length was found to be twice as long in the direction parallel to the gradient as compared to the perpendicular direction after three weeks in culture. This directionality was found to be more prominent in gradient regions of increased stiffness, crosslinked MMP-sensitive peptide presentation, and immobilized YRGDS concentration.

  16. Increased expression of matrix metalloproteinase-2 (MMP-2) predicts tumour recurrence and unfavourable outcome in non-small cell lung cancer.

    Science.gov (United States)

    Leinonen, Tero; Pirinen, Risto; Böhm, Jan; Johansson, Risto; Kosma, Veli-Matti

    2008-06-01

    The purpose of this study was to analyse the expression of matrix metalloproteinase-2 (MMP-2) and its extracellular matrix metalloproteinase inducer (EMMPRIN) in non-small cell lung cancer (NSCLC), and to evaluate their significance to predict tumour behaviour. The study consists of 212 patients treated by the resection of the tumour. Tumour samples were stained immunohistochemically, and the expression of MMP-2 and EMMPRIN was evaluated both in tumour cells and in peritumoural stromal tissue. The results were compared with clinicopathological factors and survival of the patients. High expression of MMP-2 in tumour cells was found in 83 out of 191 cases (44%). Adenocarcinomas showed more often high expression of MMP-2 as compared with squamous cell or large cell carcinomas (p=0.001). High cancer cell associated MMP-2 expression was associated with increased tumour recurrence (p=0.001). Tumour stroma showed positive staining in 162 (98%) cases and was considered highly stained in 120 (72%) cases. The high stromal MMP-2 expression was noticed more often among large cell carcinomas as compared with other histological types (p=0.007). High cancer cell associated EMMPRIN expression was found in 115 (61%) cases and was associated only with high MMP-2 expression in tumour cells (p=0.006). In overall survival (OS) and disease free survival (DFS) analyses, type of tumour (p=0.001 and p=0.0004), advanced stage (p=0.001 and p=0.013) and high MMP-2 expression in tumour cells (p=0.018 and p=0.001) were associated with poor survival. Also, high stromal MMP-2 expression was related to poor outcome in both OS and DFS analyses (p=0.010 and 0.045, respectively). In multivariate analysis, stromal MMP-2 expression retained its prognostic value to predict OS and DFS (p=0.028 and p=0.039, respectively), together with tumour type and stage (p=0.017, p=0.001 and p=0.021, p=0.008, respectively). The present study shows the significant prognostic value of MMP-2 in NSCLC suggesting that the

  17. Matrix metalloproteinase-2 functional promoter polymorphism G1575A is associated with elevated circulatory MMP-2 levels and increased risk of cardiovascular disease in systemic lupus erythematosus patients.

    Science.gov (United States)

    Bahrehmand, F; Vaisi-Raygani, A; Kiani, A; Rahimi, Z; Tavilani, H; Navabi, S J; Shakiba, E; Hassanzadeh, N; Pourmotabbed, T

    2012-05-01

    Matrix metalloproteinase-2 (MMP-2) is a zinc dependent endonuclease that degrades type IV collagen, the major structural component of basement membranes. MMP-2 functional promoter polymorphism G1575A affects circulating level of MMP-2 and may be considered an important genetic determinant of cardiovascular disease (CVD) in systemic lupus erythematosus (SLE) patients. In this study, association between MMP-2 1575A allele with serum MMP-2, neopterin and lipid-lipoprotein levels and with SLE and developing CVD was investigated. The present case-control study consisted of 109 SLE patients with and without CVD (mean age, 35.6 years) and 101 gender- and age-matched, unrelated, healthy controls (mean age, 37.1 years) from the population in the west of Iran. MMP-2 1575G/A polymorphism was detected by polymerase chain reaction (restriction fragment length polymorphism) PCR-RFLP, serum MMP-2, neopterin and lipid levels were determined by enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC) and enzyme assay, respectively. The presence of MMP-2 G1575A allele was found to be associated with SLE and developed CVD (OR = 1.78, p = 0.029 and OR = 3.43, p = 0.025, respectively). The SLE patients with MMP-2 A (G/A + A/A) allele had higher MMP-2 activity (301 ± 166 vs. 194 ± 35.5, p = 0.002), neopterin (29.4 ± 39.4 vs. 7.3 ± 4.6, p = 0.005), LDL-C (120 ± 25.7 vs. 87 ± 39.3, p = 0.045) and lower HDL-C (39.6 ± 11 vs. 45.9 ± 11.8, p = 0.031) levels than the control subjects. There was a significantly positive correlation between MMP-2 level with neopterin, total cholesterol and TG levels and negative correlation with HDL-C level in SLE patients with CVD. MMP-2 G1575A allele may be a risk factor for SLE. The carriers of this allele have high levels of MMP-2, neopterin, total cholesterol and TG and lower levels of HDL, thus, they are more likely to develop heart disease.

  18. Anti-photoaging activity and inhibition of matrix metalloproteinase (MMP) by marine red alga, Corallina pilulifera methanol extract

    Science.gov (United States)

    Ryu, BoMi; Qian, Zhong-Ji; Kim, Moon-Moo; Nam, Ki Wan; Kim, Se-Kwon

    2009-02-01

    Matrix metalloproteinases (MMPs), a key component in photoaging of the skin due to exposure to ultraviolet A, appear to be increased by UV-irradiation-associated generation of reactive oxygen species (ROS). In this study, the alga Corallina pilulifera methanol (CPM) extract has been shown to exert a potent antioxidant activity and protective effect on UVA-induced oxidative stress of human dermal fibroblast (HDF) cell. Antioxidant evaluated by various antioxidant assays. These include reducing power, total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging and protective effect on DNA damage caused by hydroxyl radicals generated. Further, the ROS level was detected using a fluorescence probe, 2',7'-dichlorofluorescein diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on HT-1080 cells. Those various antioxidant activities were compared to standard antioxidants such as α-tocopherol. In addition, the in vitro activities of MMP-2 and MMP-9 in HDF cell were inhibited by C. pilulifera methanol extract dose dependently by using gelatin zymography method. The results obtained in the present study suggested that the C. pilulifera methanol extract may be a potential source of natural anti-photoaging.

  19. Histone deacetylase (HDAC) 10 suppresses cervical cancer metastasis through inhibition of matrix metalloproteinase (MMP) 2 and 9 expression.

    Science.gov (United States)

    Song, Chenlin; Zhu, Songcheng; Wu, Chuanyue; Kang, Jiuhong

    2013-09-27

    Aberrant expression of histone deacetylases (HDACs) is associated with carcinogenesis. Some HDAC inhibitors are widely considered as promising anticancer therapeutics. A major obstacle for development of HDAC inhibitors as highly safe and effective anticancer therapeutics is that our current knowledge on the contributions of different HDACs in various cancer types remains scant. Here we report that the expression level of HDAC10 was significantly lower in patients exhibiting lymph node metastasis compared with that in patients lacking lymph node metastasis in human cervical squamous cell carcinoma. Forced expression of HDAC10 in cervical cancer cells significantly inhibited cell motility and invasiveness in vitro and metastasis in vivo. Mechanistically, HDAC10 suppresses expression of matrix metalloproteinase (MMP) 2 and 9 genes, which are known to be critical for cancer cell invasion and metastasis. At the molecular level, HDAC10 binds to MMP2 and -9 promoter regions, reduces the histone acetylation level, and inhibits the binding of RNA polymerase II to these regions. Furthermore, an HDAC10 mutant lacking histone deacetylase activity failed to mimic the functions of full-length protein. These results identify a critical role of HDAC10 in suppression of cervical cancer metastasis, underscoring the importance of developing isoform-specific HDAC inhibitors for treatment of certain cancer types such as cervical squamous cell carcinoma.

  20. Anti-photoaging activity and inhibition of matrix metalloproteinase (MMP) by marine red alga, Corallina pilulifera methanol extract

    Energy Technology Data Exchange (ETDEWEB)

    Ryu, Bo Mi [Department of Chemistry, Pukyoung National University, Busan 608-737 (Korea, Republic of); Qian Zhongji [Marine Bioprocess Research Center, Pukyong National University, Busan 608-737 (Korea, Republic of); Kim, Moon-Moo [Department of Chemistry, Dong-Eui University, Busan 614-714 (Korea, Republic of); Nam, Ki Wan [Department of Marine Biology, Pukyong National University, Busan 608-737 (Korea, Republic of); Kim, Se-Kwon [Department of Chemistry, Pukyoung National University, Busan 608-737 (Korea, Republic of); Marine Bioprocess Research Center, Pukyong National University, Busan 608-737 (Korea, Republic of)], E-mail: sknkim@pknu.ac.kr

    2009-02-15

    Matrix metalloproteinases (MMPs), a key component in photoaging of the skin due to exposure to ultraviolet A, appear to be increased by UV-irradiation-associated generation of reactive oxygen species (ROS). In this study, the alga Corallina pilulifera methanol (CPM) extract has been shown to exert a potent antioxidant activity and protective effect on UVA-induced oxidative stress of human dermal fibroblast (HDF) cell. Antioxidant evaluated by various antioxidant assays. These include reducing power, total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging and protective effect on DNA damage caused by hydroxyl radicals generated. Further, the ROS level was detected using a fluorescence probe, 2',7'-dichlorofluorescein diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on HT-1080 cells. Those various antioxidant activities were compared to standard antioxidants such as {alpha}-tocopherol. In addition, the in vitro activities of MMP-2 and MMP-9 in HDF cell were inhibited by C. pilulifera methanol extract dose dependently by using gelatin zymography method. The results obtained in the present study suggested that the C. pilulifera methanol extract may be a potential source of natural anti-photoaging.

  1. Effects of FuMing Tablets on Expression of MMP-2 in Retinal Tissue After Retinal Detachment of Rabbit%复明片对兔视网膜脱离后视网膜组织中基质金属蛋白酶-2表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘娉; 彭俊; 彭清华; 姚小磊

    2011-01-01

    Objective: To investigate the effects of FuMing Tablets (FMT) of qi - benefitting, yin - fostering, and blood - invigorating and diuresis - activating on the expression of MMP - 2 in the retinal tissue after detachment of the retina of rabbit. Methods: 36 rabbits were divided into normal contrast group (group A) , retinal detachment model group ( group B), western medicine contrast group ( group C) , FMT group( group D). Each group had 9 rabbits. Group B, C, D were made retinal detachment model. After models were made one day they were given medicine. Group A and B were administrated intragastrically with 5ml/kg warm water, once a day. Group C was administrated intragastrically with 5ml/ kg western medicine solution, once a day. Group D was administrated intragastrically with 5ml/kg FMT suspension, once a day. Then adopting the method s - p to investigate the expression of Matrix Metallo - proteinase - 2, MMP - 2 in the retinal tissue respectively in the 7th day, 14th day and 21st day. Results: MMP -2 takes on strong positive expression in the RPE ceils of model group and west medicine group, dyeing dark or pretty dark color; while weak masculine expression in the RPE cells of FMP group, dyeing shallow color. Conclusions: FMT with the functions of benefiting qi, nourishing yin, activating blood and draining water, can decrease the expression of MMP- 2 in the retinal tissue.%目的:研究益气养阴活血利水之复明片对兔实验性视网膜脱离后视网膜组织中MMP-2表达的影响.方法:将36只有色家免随机分为正常对照组(A组)、模型组(B组)、西药对照组(C组)、复明片组(D组),每组9只.B、C、D组制作视网膜脱离模型.造模后第1d下午开始给药,A组与B组均予温开水灌胃,5mL/kg体重,1次/d;C组予西药混合溶液灌胃,5ml/kg体重,1次/d;D组予复明片混悬液灌胃,5mL/kg体重,1次/d.并于造模后第7、14、21天采用S-P免疫组化法检测视网膜组织中基质金属蛋白酶-2(Matrix

  2. 异丙酚对肺气肿大鼠 MMP-2和 MMP-9的影响%Effect of propofol on matrix metaloproteinase-2 and matrix metaloproteinase-9 in emphysema in rats

    Institute of Scientific and Technical Information of China (English)

    闵振兴; 陈丽辉; 黄漫

    2013-01-01

      Objective: To study the Effect of propofol on matrix metaloproteinase-2 and matrix metaloproteinase-9 in emphysema in rats and its clinical significance. Methods:30 Wistar rats, weighing 200 to 250 g, male or female. Were randomly divided into three groups: smoking plus Klebsiela pneumoniae infection group (S group), propofol treatment group P group and the control group (C group). (N = 10). S Group and P Group were infected Klebsiela pneumoniae contamination by nasal inhalation 1-2 times a week, each time each animal 0.1ml, a total of 10 weeks; both groups were inhaled cigarettes through the catheter each 2h, 5 times a week for 16 C Group animals were no smoke, while the culture medium instead of bacteria inhaled.RESULTS: Under light microscope, C group lung tissue was normal, But S group showed severe structural damage, alveolar colapse, interstitial pulmonary edema, hemorrhage, inflammatory cel infiltration, showed obvious signs of emphysema changes. Pathological changes of the P group were light compared with S group. Conclusions: MMP-2 and MMP-9 expression are significantly increased in emphysema in rats, and propofol can protect lung function in related to inhibit matrix metaloproteinases.%  目的探索异丙酚对肺气肿大鼠血清和气管肺泡灌洗液 MMP-2和 MMP-9的变化及其临床意义。方法Wistar 大鼠30只,体重200-250 g,雌雄不拘。分为三组:吸烟加肺炎克雷伯杆菌感染组(S 组)、异丙酚处理组(P 组)和对照组(C 组)。每组10只。S 组和 P 组采用经鼻吸入肺炎克雷伯杆菌感染的方法染菌,每周1-2次,每次每只动物0.1ml,共10周;两组香烟吸入每次使用12支,分4回燃烧,烟雾通过导管进入约300L 的烟雾吸入箱内,每次2h,每周5次,共16周。C 组动物不吸烟,以培养液代替菌液经鼻吸入。结果光镜下 C 组肺组织结构正常,S 组肺组织呈现结构毁损严重,肺泡塌陷,肺间质水肿,出血,炎症细胞浸润,呈

  3. MicroRNA miR-182 cluster mediated modulation of RECK without changes in cell surface membrane type-1 matrix metalloproteinase (MT1-MMP).

    Science.gov (United States)

    Silva, Milagros; Hernandez, Maria E; Rojas, Fausto; Li, Lihua; Subramanian, Subbaya; Wilson, Michael J

    2015-01-01

    Cell surface localized membrane type 1-matrix metalloproteinase (MT1-MMP) plays an important role in physiological and pathological processes and its function can be regulated by proteins such as RECK. We examined the ability of miR-182 (one of the miR-183 cluster miRNAs), which can target RECK, to control cell surface MT1-MMP activity. Expression of RECK mRNA and protein was increased with anti-miRs to miR-182, miR-183 or miR-96 in HT1080 fibrosarcoma cells, but, decreased RECK mRNA and increased its protein in the benign prostatic hyperplasia cell line BPH-1. Treatment of BPH-1 and HT-1080 cells with the anti-miRs did not change the level of cell surface MT1-MMP activity, nor their rate of migration in an in vitro wound-healing assay. Trichostatin A (TSA) did not increase the level of RECK, but blocked cell surface MT1-MMP activity and decreased cell motility. Anti-miRs mediated increased RECK levels did not interfere with cell surface MT1-MMP function, and TSA may block cell surface localization of MT1-MMP by a mechanism independent of RECK.

  4. Enhancement of Matrix Metalloproteinase-2 (MMP-2) as a Potential Chondrogenic Marker during Chondrogenic Differentiation of Human Adipose-Derived Stem Cells.

    Science.gov (United States)

    Arai, Yoshie; Park, Sunghyun; Choi, Bogyu; Ko, Kyoung-Won; Choi, Won Chul; Lee, Joong-Myung; Han, Dong-Wook; Park, Hun-Kuk; Han, Inbo; Lee, Jong Hun; Lee, Soo-Hong

    2016-06-17

    Human adipose-derived stem cells (hASCs) have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation. Recently, hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time. Matrix metalloproteinases (MMPs) are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells. MMPs secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs.

  5. Leukolysin/MMP25/MT6—MMP:a novel matrix metalloproteinase specifically expressed in the leukocyte lineage

    Institute of Scientific and Technical Information of China (English)

    PEIDUANQING

    1999-01-01

    A novel matrix matalloproteinase (MMP) was identified from leukocytes and found to be specifically expressed by peripheral blood leukocytes among 29 different tissues examined.Named leukolysin,it encodes for 562 residues with a conserved MMP structure,i.e.,pre-,pro-,catalytic,hinge- and hemopexin-like domains,but also a RXK/RR motif,known for its role in MMP zymogen activation,and a C-terminal hydrophobic segment.Overall,leukolysin displays the strongest homology to the newly identified MT-MMP subgroup with 45% and 39% identities to MT4- and MT1-MMPs vs 30% and 31.5% to MMP1 and 3 respectively.Unlike MT4-MMP whose proteolytic activity remains undefined,a C-terminally truncated leukolysin is expressed as a strong gelatinolytic species at 28kDa which is derived from a cell-associated 34kDa proenzyme,presumably by furin or proprotein convertase mediated removal of the propeptide(-6kDa).By green fluorescent protein(GFP) tagging,the intracellular groenzyme is localized to granules throughout the cell,suggesting that activation occur immediately prior to secretion.Taken together,leukolysin may be part of the proteolytic arsenal deployed by leukocytes during inflammatory responses.

  6. Enhancement of Matrix Metalloproteinase-2 (MMP-2 as a Potential Chondrogenic Marker during Chondrogenic Differentiation of Human Adipose-Derived Stem Cells

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    Yoshie Arai

    2016-06-01

    Full Text Available Human adipose-derived stem cells (hASCs have a capacity to undergo adipogenic, chondrogenic, and osteogenic differentiation. Recently, hASCs were applied to various fields including cell therapy for tissue regeneration. However, it is hard to predict the direction of differentiation of hASCs in real-time. Matrix metalloproteinases (MMPs are one family of proteolytic enzymes that plays a pivotal role in regulating the biology of stem cells. MMPs secreted by hASCs are expected to show different expression patterns depending on the differentiation state of hASCs because biological functions exhibit different patterns during the differentiation of stem cells. Here, we investigated proteolytic enzyme activity, especially MMP-2 activity, in hASCs during their differentiation. The activities of proteolytic enzymes and MMP-2 were higher during chondrogenic differentiation than during adipogenic and osteogenic differentiation. During chondrogenic differentiation, mRNA expression of MMP-2 and the level of the active form of MMP-2 were increased, which also correlated with Col II. It is concluded that proteolytic enzyme activity and the level of the active form of MMP-2 were increased during chondrogenic differentiation, which was accelerated in the presence of Col II protein. According to our findings, MMP-2 could be a candidate maker for real-time detection of chondrogenic differentiation of hASCs.

  7. MATRIX METALLOPROTEINS (MMP)-MEDIATED PHOSPHORYLATION OF THE EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR) IN HUMAN AIRWAY EPITHELIAL CELLS (HAEC) EXPOSED TO ZINC (ZN)

    Science.gov (United States)

    Matrix Metalloproteinase (MMP)-Mediated Phosphorylation of The Epidermal Growth Factor Receptor (EGFR) in Human Airway Epithelial Cells (HAEC) Exposed to Zinc (Zn)Weidong Wu, James M. Samet, Robert Silbajoris, Lisa A. Dailey, Lee M. Graves, and Philip A. BrombergCenter fo...

  8. Direct inhibition of matrix metalloproteinase-2 (MMP-2) by (-)-epigallocatechin-3-gallate: A possible role for the fibronectin type II repeats.

    Science.gov (United States)

    Jha, Shalinee; Kanaujia, Shankar Prasad; Limaye, Anil M

    2016-11-15

    Matrix metalloproteinases (MMPs) -2 and -9, also called gelatinases, constitute a distinct subgroup within the MMP family of extracellular matrix remodeling proteases. Gelatinases are implicated in tumor cell invasion and metastasis, and are attractive therapeutic targets. Several synthetic small molecule inhibitors of MMPs developed till date have failed in clinical trials. This has prompted explorations into the gamut of dietary compounds and nutraceuticals for specific inhibitors of MMPs with desirable properties. (-)-epigallocatechin-3-gallate (EGCG), a major green tea polyphenol, is popular as a potential chemotherapeutic agent with demonstrable anti-metastatic and MMP inhibitory activities. Here, we have addressed the mechanism of EGCG-mediated inhibition of MMP-2 using in silico molecular docking approach. We show for the first time that EGCG targets the fibronectin type II repeat regions 1 and 3 of MMP-2, binds amino acids that constitute the exosite of this enzyme and hinders proper positioning of the substrate. This study offers a novel insight into the inhibition of MMP-2 by EGCG and presents a starting point for development of novel therapeutic molecules that can specifically target the gelatinases.

  9. Matrix metalloproteinases (MMP) and cathepsin K contribute differently to osteoclastic activities

    DEFF Research Database (Denmark)

    Delaissé, Jean-Marie; Andersen, Thomas L; Engsig, Michael T;

    2003-01-01

    is based on a model of osteoclast recruitment in primitive long bones, an assay of osteoclast invasion through collagen gel, and the effect of proteinase inhibitors/knockouts in these models. Furthermore, we mention observations indicating a role of MMPs in initiation of bone resorption. Finally, we......The best established proteolytic event of osteoclasts is bone matrix solubilization by the cysteine proteinase cathepsin K. Here, however, we draw the attention on osteoclastic activities depending on matrix metalloproteinases (MMPs). We discuss the observations supporting that MMPs contribute...

  10. Thermodynamic analysis of ionizable groups involved in the catalytic mechanism of human matrix metalloproteinase 7 (MMP-7).

    Science.gov (United States)

    Takeharu, Hitoshi; Yasukawa, Kiyoshi; Inouye, Kuniyo

    2011-12-01

    Human matrix metalloproteinase 7 (MMP-7) exhibits a broad bell-shaped pH-dependence with the acidic and alkaline pK(e) (pK(e1) and pK(e2)) values of about 4 and 10. In this study, we estimated the ionizable groups involved in its catalytic mechanism by thermodynamic analysis. pK(a) of side chains of L-Asp, L-Glu, L-His, L-Cys, L-Tyr, L-Lys, and L-Arg at 25-45°C were determined by the pH titration of amino-acid solutions, from which their enthalpy changes, ∆H°, of deprotonation were calculated. pK(e1) and pK(e2) of MMP-7 at 15-45°C were determined in the hydrolysis of (7-methoxycoumarin-4-yl)acetyl-L-Pro-L-Leu-Gly-L-Leu-[N(3)-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH(2), from which ∆H(o) for pK(e1) and pK(e2) was calculated. The ∆H(o) for pK(e1) (-20.6±6.1kJmol(-1)) was similar to that for L-Glu (-23.6±5.8kJmol(-1)), and the ∆H(o) for pK(e2) (89.9±4.0kJmol(-1)) was similar to those for L-Arg (87.6±5.5kJmol(-1)) and L-Lys (70.4±4.4kJmol(-1)). The mutation of the active-site residue Glu198 into Ala completely abolished the activity, suggesting that Glu198 is the ionizable group for pK(e1). On the other hand, no arginine or lysine residues are found in the active site of MMP-7. We proposed a possibility that a protein-bound water is the ionizable group for pK(e2).

  11. Increased soluble CD4 in serum of rheumatoid arthritis patients is generated by matrix metalloproteinase (MMP-like proteinases.

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    Wen-Yi Tseng

    Full Text Available Higher soluble CD4 (sCD4 levels in serum have been detected in patients of infectious and chronic inflammatory diseases. However, how and why sCD4 is produced remains poorly understood. We establish sensitive ELISA and WB assays for sCD4 detection in conditioned medium of in vitro cell culture system and serum of chronic inflammatory patients. Serum samples from patients with systemic lupus erythematosus (SLE (n = 79, rheumatoid arthritis (RA (n = 59, ankylosing spondylitis (AS (n = 25, gout (n = 31, and normal controls (n = 99 were analyzed using ELISA for sCD4 detection. Results from each assay were analyzed by the Kruskal-Wallis test. Dunn's multiple comparison post-test was then applied between groups. We confirm that cells expressing exogenous CD4 produce sCD4 in a constitutive and PMA-induced manner. Importantly, sCD4 production in a heterologous expression system is inhibited by GM6001 and TAPI-0, suggesting receptor shedding by matrix metalloproteinase (MMP-like proteinases. Moreover, similar findings are recapitulated in human primary CD4(+ T cells. Finally, we show that serum sCD4 levels are increased in patients of chronic inflammatory diseases including RA and SLE, but not in those with gout. Intriguingly, sCD4 levels in RA patients are correlated positively with the disease activities and higher sCD4 levels seem to associate with poor prognosis. Taken together, we conclude that CD4 is shed from cell surface by a MMP-like sheddase and sCD4 level is closely related with the inflammatory condition in certain chronic diseases. Hence, sCD4 might be considered an important parameter for RA disease progression with potential diagnostic importance.

  12. Use of matrix metalloproteinase-9 (MMP-9 and its tissue inhibitor (TIMP-1 in the pathomorphological diagnosis of carotid pathology: literature review and own observations

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    Yu. I. Kuzyk

    2016-01-01

    Full Text Available Matrix metalloproteinases (MMPs are the degradative enzymes of the extracellular matrix. Currently, the role of MMP-2 and MMP-9 in the progression of atherosclerosis (AS is proved. The question of possible involvement of MMP-9 into elastin degradation in fibromuscular dysplasia (FMD and pathological tortuosity (PT remains open and insufficiently explored. The aim of the study – analysis of the current literature on the role of degradative enzymes in the development of carotid pathology and study of the expression of type I, III, IV collagens, MMP-9 and TIPM-1 in the wall of the carotid arteries in FMD, PT and AS. Materials and methods included literature review and own research. Immunohistochemical study of type I, III and IV collagens, TIMP-1 and MMP-9 was carried out on surgical material of patients with main carotid diseases: three observations with AS, two – with FMD, two – with PT. The level of expression was assessed by semiquantitative method. Results. Own observations showed that in FMD types I and III collagen content in the media and in the adventitia remains unchanged. MMP-9 expression level reached the highest level of intensity in atherosclerotic plaques, particularly in macrophages, constituting the main part of the atheromatous mass. Moderate intensity of expression is noted in FMD and PT. In PT expression prevailed in the lower third of the media on the border with adventitia, including the adventitia, in FMD – mainly in the media. The level of TIMP-1 is weakly positive in PT and FMD, negative in AS. Conclusions. These results demonstrate the possibility of using MMP-9 and TIMP-1 as a morphological marker determining pathological processes in carotid pathology. Data of immunohistochemical study of type I, II, IV collagens indicate moderate expression of collagen type I in FMD and PT, severe expression of collagen III in FMD, moderate in PT. Type IV collagen is highly expressed in atherosclerotic plaques. For AS high

  13. Matrix metalloproteinase (MMP-9 in cancer-associated fibroblasts (CAFs is suppressed by omega-3 polyunsaturated fatty acids in vitro and in vivo.

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    Ayumi Taguchi

    Full Text Available Cancer associated fibroblasts (CAFs are responsible for tumor growth, angiogenesis, invasion, and metastasis. Matrix metalloproteinase (MMP-9 secreted from cancer stroma populated by CAFs is a prerequisite for cancer angiogenesis and metastasis. Omega-3 polyunsaturated fatty acids (omega-3 PUFA have been reported to have anti-tumor effects on diverse types of malignancies. Fat-1 mice, which can convert omega-6 to omega-3 PUFA independent of diet, are useful to investigate the functions of endogenous omega-3 PUFA. To examine the effect of omega-3 PUFA on tumorigenesis, TC-1 cells, a murine epithelial cell line immortalized by human papillomavirus (HPV oncogenes, were injected subcutaneously into fat-1 or wild type mice. Tumor growth and angiogenesis of the TC-1 tumor were significantly suppressed in fat-1 compared to wild type mice. cDNA microarray of the tumors derived from fat-1 and wild type mice revealed that MMP-9 is downregulated in fat-1 mice. Immunohistochemical study demonstrated immunoreactivity for MMP-9 in the tumor stromal fibroblasts was diffusely positive in wild type whereas focal in fat-1 mice. MMP-9 was expressed in primary cultured fibroblasts isolated from fat-1 and wild type mice but was not expressed in TC-1 cells. Co-culture of fibroblasts with TC-1 cells enhanced the expression and the proteinase activity of MMP-9, although the protease activity of MMP-9 in fat-1-derived fibroblasts was lower than that in wild type fibroblasts. Our data suggests that omega-3 PUFAs suppress MMP-9 induction and tumor angiogenesis. These findings may provide insight into mechanisms by which omega-3 PUFAs exert anti-tumor effects by modulating tumor microenvironment.

  14. The role of matrix metalloproteinase MMP-9 and TIMP-2 tissue inhibitor of metalloproteinases as serum markers of bladder cancer.

    Science.gov (United States)

    Ramón de Fata, F; Ferruelo, A; Andrés, G; Gimbernat, H; Sánchez-Chapado, M; Angulo, J C

    2013-09-01

    The diagnosis and molecular staging of bladder cancer based on the detection of gelatinases mRNA (MMP-2 and MMP-9) in peripheral blood circulating and mononuclear cells have shown promising results. We analyze if the determination of the corresponding protein synthesis products makes it possible to diagnose and characterize patients with bladder cancer. Quantification of the serum levels of MMP-2, MMP-9 and TIMP-2 in a series of 42 individuals (31 patients with bladder cancer in different stages and 11 healthy controls) using the ELISA technique was carried out. The determinations were compared between cases and controls (Mann-Whitney U) and between different groups of tumors (Mann-Whitney U or Kruskal-Wallis), according to the clinical-pathological characteristics (age, gender, T category, M category or grade). Diagnostic yield of these markers was evaluated by analysis of the ROC curves. There is a correlation between the determinations of MMP-2 and TIMP-2 (R=.699; P>.0001) and MMP-9 and TIMP-2 (R=.305; P=.049). Patients with bladder cancer have higher levels of MMP-9 (p<0.0001) and TIMP-2 (P=.047) than the controls. Furthermore, the MMP-9/TIMP-2 ratio is also superior in cancer patients (P<.001). Differences were not detected between cancer and controls regarding age (P=.64) or gender (P=.64). Differences were also not detected regarding MMP-2 (P=.35) or MMP-2/TIMP-2 rate (P=.45). Within the cancer patient population, the MMP-2 and MMP-9 values differ according to T category (P=.022 and P=.038, respectively) and those of the TIMP-2 according to M category (P=.036). ROC curve analysis showed that both MMP-9 and the MMP-9/TIMP-2 ratio discriminate patients with cancer and controls, with equivalent diagnostic accuracy (ABC 0.953) and cut offs of 3.93 ng/mL (S 90%; Sp 81%) and 0.053 ng/mL (S 96%; Sp 84%), respectively. The results obtained suggest that both serum MMP-9 and TIMP-2 would have an application in the prediction of the development and progression of

  15. Matrix Metalloproteinase 2 (MMP-2) Plays a Critical Role in the Softening of Common Carp Muscle during Chilled Storage by Degradation of Type I and V Collagens.

    Science.gov (United States)

    Xu, Chao; Wang, Cheng; Cai, Qiu-Feng; Zhang, Qian; Weng, Ling; Liu, Guang-Ming; Su, Wen-Jin; Cao, Min-Jie

    2015-12-30

    Matrix metalloproteinases (MMPs) are proposed to play important roles in the degradation of collagens, thus causing the post-mortem softening of fish muscle, although the specific mechanism remains largely unresolved. Previously, we reported the existence of gelatinase-like proteinases in common carp (Cyprinus carpio) muscle. The primary structures of these proteinases, however, have never been investigated. In the present study, two MMPs with molecular masses of 66 and 65 kDa were purified to homogeneity from common carp muscle by ammonium sulfate fractionation and a series of column chromatographies. Matrix-assisted laser desorption/ionization-tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS/MS) analysis indicated that they are completely identical to MMP-2 from common carp. During chilled storage of common carp at 4 °C, the enzymatic activity of MMP-2 increased to 212% in 12 h while the texture profile increased over the first 2 h and gradually decreased. On the other hand, type V collagen was purified to homogeneity and a specific polyclonal antibody against this protein was prepared. Both type I and V collagens were effectively hydrolyzed by MMP-2 at 30 °C and even at 4 °C. Furthermore, injection of metalloproteinase proteinase inhibitor EDTA into the blood vessel of live common carp suppressed post-mortem tenderization significantly. All of these results confirmed that MMP-2 is a major proteinase responsible for the degradation of collagens, resulting in the softening of fish muscle during chilled storage.

  16. EXPRESS AND ROLE OF MATRIX METALLOPROTEINASE-2 (MMP-2)、 TISSUE INHIBITOR OF METALLOPROTEINASE-2 (TIMP-2)、 MEMBRANE TYPE 1-MATRIX METALLOPROTEINASE (MT1-MMP) mRNA IN AMELOBLASTOMA%成釉细胞瘤MMP-2、TIMP-2、MT1-MMP mRNA的表达和意义

    Institute of Scientific and Technical Information of China (English)

    张彬; 黄洪章; 潘朝斌; 陶谦

    2003-01-01

    目的探讨MMP-2、TIMP-2、MT1-MMP与成釉细胞瘤局部侵袭特性的生物学关系.方法采用RT-PCR方法检测基质金属蛋白酶及其抑制剂和激活剂MMP-2、TIMP-2、MT1-MMP在成釉细胞瘤的表达及含量.结果在成釉细胞瘤组织中,MMP-2、MT1-MMP、TIMP-2的mRNA均表达,且表达水平均显著高于正常牙囊组织,复发及实性成釉细胞瘤的TIMP-2、MT1-MMP相对含量均显著高于原发和囊性成釉细胞瘤.结论在成釉细胞瘤组织中,MMP-2、MT1-MMP、TIMP-2转录水平的增高可能与其侵袭特性有关,MT1-MMP/TIMP-2表达水平可能与其侵袭能力有关.

  17. Interleukin-4 inhibition of interleukin-1-induced expression of matrix metalloproteinase-3 (MMP-3 is independent of lipoxygenase and PPARγ activation in human gingival fibroblasts

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    Gorski Grzegorz

    2007-02-01

    Full Text Available Abstract Background Interleukin 4 (IL-4 has been shown to suppress interleukin-1 (IL-1 induced expression of matrix metalloproteinase-3 (MMP-3 in human synovial and gingival fibroblasts, but the mechanism of suppression has not been determined. Activators of peroxisome proliferator-activated receptor-γ (PPARγ have been shown to inhibit cytokine induced expression of MMPs in other cell types, and IL-4 has been shown to activate PPARγ by stimulating production of ligands through the lipoxygenase pathway. It has been suggested that PPARγ may inhibit expression of MMPs by competing with transcription factor AP-1 for binding to a putative composite binding element in the promoters. The objective of this study was to determine whether the suppressive effects of IL-4 on the IL-1 induced expression of MMP-3 involve activation of lipoxygenase and/or PPARγ. Results Western blotting revealed the presence of PPARγ in nuclear extract of HGF. IL-1 induced binding of nuclear extract to the putative composite PPRE/AP-1 site was diminished in the presence of pioglitazone, but there was no evidence of any change in the composition of the retarded complexes, and no evidence of PPARγ binding to this site. Nordihydroguaiaretic acid (NDGA, a non-selective lipoxygenase inhibitor, and MK886, a specific inhibitor of 5-lipoxygenase, induced MMP-3 expression synergistically with IL-1. However IL-4 was still able to inhibit MMP-3 expression in the presence of NDGA or MK886 and IL-1. Activation of PPARγ with pioglitazone not only failed to inhibit IL-1 induced expression of MMP-3 mRNA, but rather super-induced MMP-3 in the presence of IL-1. PPARγ antagonist GW9662 failed to abolish the suppressive effects of IL-4. Another PPARγ activator, 15-deoxy-Delta12,14prostaglandin J2 (15dPGJ2, also super-induced MMP-3 mRNA, and this was due at least in part to increased transcription. Conclusion IL-4 suppression of IL-1-induced MMP-3 expression in HGF is independent of

  18. Caffeine induces matrix metalloproteinase-2 (MMP-2) and MMP-9 down-regulation in human leukemia U937 cells via Ca2+/ROS-mediated suppression of ERK/c-fos pathway and activation of p38 MAPK/c-jun pathway.

    Science.gov (United States)

    Liu, Wen-Hsin; Chang, Long-Sen

    2010-09-01

    Caffeine attenuated invasion of human leukemia U937 cells with characteristic of decreased protein expression and mRNA levels of matrix metalloproteinase-2 (MMP-2) and MMP-9. Down-regulation of MMP-2 and MMP-9 in U937 cells was abrogated by abolishment of caffeine-elicited increase in intracellular Ca(2+) concentration and ROS generation. Pretreatment with BAPTA-AM (Ca(2+) chelator) and N-acetylcysteine (ROS scavenger) abolished caffeine-induced ERK inactivation and p38 MPAK activation. Moreover, caffeine treatment led to MAPK phosphatase-1 (MKP-1) down-regulation and protein phosphatase 2A catalytic subunit (PP2Ac) up-regulation, which were involved in cross-talk between p38 MAPK and ERK. Transfection of constitutively active MEK1 or pretreatment with SB202190 (p38 MAPK inhibitor) restored MMP-2 and MMP-9 protein expression in caffeine-treated cells. Caffeine treatment repressed ERK-mediated c-Fos phosphorylation but evoked p38 MAPK-mediated c-Jun phosphorylation. Knock-down of c-Fos and c-Jun by siRNA reflected that c-Fos counteracted the effect of c-Jun on MMP-2/MMP-9 down-regulation. Taken together, our data indicate that MMP-2/MMP-9 down-regulation in caffeine-treated U937 cells is elicited by Ca(2+)/ROS-mediated suppression of ERK/c-Fos pathway and activation of p38 MAPK/c-Jun pathway.

  19. Galectin-3 facilitates cell motility in gastric cancer by up-regulating protease-activated receptor-1 (PAR-1 and matrix metalloproteinase-1 (MMP-1.

    Directory of Open Access Journals (Sweden)

    Seok-Jun Kim

    Full Text Available BACKGROUND: Galectin-3 is known to regulate cancer metastasis. However, the underlying mechanism has not been defined. Through the DNA microarray studies after galectin-3 silencing, we demonstrated here that galectin-3 plays a key role in up-regulating the expressions of protease-activated receptor-1 (PAR-1 and matrix metalloproteinase-1 (MMP-1 PAR-1 thereby promoting gastric cancer metastasis. METHODOLOGY/PRINCIPAL FINDINGS: We examined the expression levels of Galectin-3, PAR-1, and MMP-1 in gastric cancer patient tissues and also the effects of silencing these proteins with specific siRNAs and of over-expressing them using specific lenti-viral constructs. We also employed zebrafish embryo model for analysis of in vivo gastric cancer cell invasion. These studies demonstrated that: a galectin-3 silencing decreases the expression of PAR-1. b galectin-3 over-expression increases cell migration and invasion and this increase can be reversed by PAR-1 silencing, indicating that galectin-3 increases cell migration and invasion via PAR-1 up-regulation. c galectin-3 directly interacts with AP-1 transcriptional factor, and this complex binds to PAR-1 promoter and drives PAR-1 transcription. d galectin-3 also amplifies phospho-paxillin, a PAR-1 downstream target, by increasing MMP-1 expression. MMP-1 silencing blocks phospho-paxillin amplification and cell invasion caused by galectin-3 over-expression. e Silencing of either galectin-3, PAR-1 or MMP-1 significantly reduced cell migration into the vessels in zebrafish embryo model. f Galectin-3, PAR-1, and MMP-1 are highly expressed and co-localized in malignant tissues from gastric cancer patients. CONCLUSIONS/SIGNIFICANCE: Galectin-3 plays the key role of activating cell surface receptor through production of protease and boosts gastric cancer metastasis. Galectin-3 has the potential to serve as a useful pharmacological target for prevention of gastric cancer metastasis.

  20. Expression of matrix metalloproteinase 2 and 9 in hypertension-induced intracranial aneurysm in rats%实验性大鼠脑动脉瘤形成过程中MMP-2MMP-9表达的动态变化

    Institute of Scientific and Technical Information of China (English)

    马良; 付强; 关俊宏

    2013-01-01

    目的 探讨实验性大鼠动脉瘤形成过程中基质金属蛋白酶2 (MMP-2)、基质金属蛋白酶9(MMP-9)的表达规律.方法 制作肾性高血压大鼠脑动脉瘤模型,通过免疫组化在蛋白水平系统动态观察肾性高血压大鼠脑动脉瘤形成过程中MMP-2、MMP-9表达的变化.结果 实验组在术后1 w脑动脉壁即可见MMP-2、MMP-9表达增加,随着术后时间的推移和大鼠血压的增高,其表达也迅速增加,术后1个月基本达最高峰并一直持续至4个月,其中MMP-9较正常状态的增加比MMP-2的表达增加更为显著.对照组脑动脉壁MMP-2、MMP-9也有微弱表达,且MMP-2表达较MMP-9略强.结论 脑动脉壁MMP-9、MMP-2特别是MMP-9的过度表达导致的脑动脉壁胶原纤维及内弹力层破坏是脑动脉瘤形成的主要原因之一.%Objective To investigate the expression pattern of matrix metalloproteinase-2 (MVP-2),matnx metalloproteinase-9 (MMP-9) in the process of aneurysm formation in rats.Methods A rat model of cerebral aneurysm was established.The dynamic changes of MMP-2 and MMP-9 expressions were examined by irrmmunohistochemistry at the level of protein activity in the process of aneurysm formation.Results In experimental group,with the development of hypertension,the expressions of MMP-2 and MMP-9 were increased one week after operation,peaked at first month and lasted until 4 months after operation.The expression of MMP-9 was increased more remarkably than that of MMP-2.Weak staining of MMP-2 and MMP-9 was observed in the cerebral artery,and the expression of MMP-2 was little higher than that of MMP-9.Conclusion The over-expressions of MMP-2 and MMP-9 in artery which result in the wall destruction of collagen and internal elastic lamina of cerebral artery are the main cause of the aneurysm formation.

  1. Polymorphisms in matrix metalloproteinases MMP1 and MMP9 are associated with primary open-angle and angle closure glaucoma in a Pakistani population

    NARCIS (Netherlands)

    Micheal, S.; Yousaf, S.; Khan, M.I.; Akhtar, F.; Islam, F.; Khan, W.A.; Hollander, A.I. den; Qamar, R.; Ahmed, A.

    2013-01-01

    PURPOSE: Matrix metalloproteinases (MMPs) play an important role in remodeling of the extracellular matrix during development and growth of various tissues including the eye. Various functional polymorphisms in MMPs have been implicated in the pathogenesis of different types of glaucoma. The aim of

  2. Thromboxane A2 receptor-mediated release of matrix metalloproteinase-1 (MMP-1) induces expression of monocyte chemoattractant protein-1 (MCP-1) by activation of protease-activated receptor 2 (PAR2) in A549 human lung adenocarcinoma cells.

    Science.gov (United States)

    Li, Xiuling; Tai, Hsin-Hsiung

    2014-08-01

    Matrix metalloproteinases (MMPs) and monocyte chemoattractant protein-1 (MCP-1, CCL2) are known to be upregulated in many tumors. Their roles in tumor invasion and metastasis are being uncovered. How they are related to each other and involved in tumor progression remains to be determined. Earlier it was reported that I-BOP-initiated activation of thromboxane A2 receptor (TP) induced the release of MMP-1, MMP-3, and MMP-9 from lung cancer A549 cells overexpressing TPα (A549-TPα). Herein it was found that MMP-1, but not MMP-3 or MMP-9, induced the expression of MCP-1 in A549 cells. Conditioned medium (CM) from I-BOP activated, MMP-1 siRNA pretreated A549-TPα cells induced greatly attenuated expression of MCP-1 in A549 cells indicating that MMP-1 in the CM contributed significantly to the expression of MCP-1. MMP-1 was shown to activate protease-activated receptor 2 (PAR2) instead of commonly assumed PAR1 to increase the expression of MCP-1 in A549 cells. This conclusion was reached from the following findings: (1) expression of MCP-1 induced by trypsin, a PAR2 agonist, and also PAR2 agonist peptide, was inhibited by a PAR2 antagonist; (2) expression of MCP-1 induced by MMP-1 and by CM from I-BOP activated A549-TPα cells was blocked by a PAR2 antagonist but not by other PAR antagonists; (3) expression of MCP-1 induced by MMP-1 and by CM from I-BOP activated A549-TPα cells was attenuated significantly by pretreatment of cells with PAR2-siRNA. These results suggest that PAR2 is a novel MMP-1 target mediating MMP-1-induced signals in A549 lung cancer cells.

  3. Synthesis, radiosynthesis, in vitro and preliminary in vivo evaluation of biphenyl carboxylic and hydroxamic matrix metalloproteinase (MMP) inhibitors as potential tumor imaging agents

    Energy Technology Data Exchange (ETDEWEB)

    Oltenfreiter, Ruth [Laboratory of Radiopharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent (Belgium)]. E-mail: ruth.oltenfreiter@ugent.be; Staelens, Ludovicus [Laboratory of Radiopharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent (Belgium); Hillaert, Ulrik [Laboratory for Medicinal Chemistry, Ghent University, Harelbekestraat 72, 9000 Ghent (Belgium); Heremans, An; Noel, Agnes [Laboratory of Tumor and Developmental Biology, University of Liege, Sart-Tilman, Liege (Belgium); Frankenne, Francis [Laboratory of Tumor and Developmental Biology, University of Liege, Sart-Tilman, Liege (Belgium); Slegers, Guido [Laboratory of Radiopharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent (Belgium)

    2005-06-01

    Excess matrix degradation is one of the hallmarks of cancer and is an important factor in the process of tumor progression. It is implicated in invasion, metastasis, growth, angiogenesis and migration. Many characteristics of matrix metalloproteinases (MMPs) make them attractive therapeutic and diagnostic targets. MMP expression is upregulated at the tumor site, with localization of activity in the tumor or the surrounding stroma, providing a target for medical imaging techniques. Radioiodinated carboxylic and hydroxamic MMP inhibitors 2-(4'-[{sup 123}I] iodo-biphenyl-4-sulfonylamino)-3-methyl-butyric acid (9) and 2-(4'-[{sup 123}I] iodo-biphenyl-4-sulfonylamino)-3-methyl-butyramide (11), their unlabelled standards and precursors were synthesized. Radioiodination was conducted by electrophilic aromatic substitution of the tributylstannyl precursors and resulted in radiochemical yields of 70+/-5% (n=6) and 60+/-5% (n=4), respectively. In vitro zymography and enzyme assays showed for both hydroxamic acid and carboxylic acid compounds a good inhibition activity and a high selectivity for MMP-2. In vivo biodistribution in NMRI mice showed no long-term accumulation in organs and the possibility to accumulate in the tumor in a later phase of this study.

  4. 急性视网膜坏死患者玻璃体MMP-2和MMP-9活性研究%Investigation of the activity of matrix metalloproteinases 2 and 9 in vitreous of patients with acute retinal necrosis

    Institute of Scientific and Technical Information of China (English)

    顾永昊; 石磊; 柯根杰

    2009-01-01

    目的 检测急性视网膜坏死(acute retinal necrosis,ARN)患者玻璃体内基质金属蛋白酶(matrix metallopreteinases,MMPs)2和9的活性.方法 6例ARN患者行玻璃体切割术抽取玻璃体液,6例单纯孔源性视网膜脱离(rhegmatogenous detachment,RD)患者作为对照.采用明胶酶谱法进行MMP-2和MMP-9活性检测.结果 RD患者玻璃体标本显示72kDaMMP-2酶原和62kDa MMP-2活性酶条带,无明显MMP-9条带成分,ARN患者标本可见到92kDaMMP-9酶原、82kDaMMP-9活性酶、72kDaMMP-2酶原和62kDa MMP-2活性酶条带,ARN组所有条带的表达都要显著高于RD组(P<0.01).结论 MMP-2和MMP-9可能在ARN的玻璃体视网膜炎症反应中起到非常重要的作用.%Objective To detect the activity of matrix metalloproteinases (MMP) 2 and 9 in vitreous of patients with acute retinal necrosis (ARN). Methods Vitreous samples from 6 cases with ARN were obtained in process of vitrectomy. Samples from 6 cases of rheg-matogenous detachment of retina (RD) without PVR were set as control. Activities of MMP-2 and-9 were detected by gelatin zymography. Re-stilts Only 72kDa zemogen and 62 active enzyme of MM-2 could be detected in control, while 92kDa zemogen and 82 active enzyme of MM-9 and 72kDa zemogen and 62 active enzyme of MM-2 could be detected in vitreous of ALAN. Concentrations of all four bands were significantly higher in ARN patients than that in control (P<0.01). Conclusion Gelatinase A (MMP-2) and B (MMP-9) might play important roles in in-flammatory responses of ALAN patients.

  5. Fetal insulin and IGF-II contribute to gestational diabetes mellitus (GDM)-associated up-regulation of membrane-type matrix metalloproteinase 1 (MT1-MMP) in the human feto-placental endothelium.

    Science.gov (United States)

    Hiden, U; Lassance, L; Tabrizi, N Ghaffari; Miedl, H; Tam-Amersdorfer, C; Cetin, I; Lang, U; Desoye, G

    2012-10-01

    Gestational diabetes mellitus (GDM)-associated hormonal and metabolic derangements in mother and fetus affect placental development and function. Indeed, in GDM, placentas are characterized by hypervascularization and vascular dysfunction. The membrane-type matrix metalloproteinase 1 (MT1-MMP) is a key player in angiogenesis and vascular expansion. Here, we hypothesized elevated placental MT1-MMP levels in GDM induced by components of the diabetic environment. Therefore, we measured placental MT1-MMP in normal vs. GDM pregnancies, identified potential functional consequences, and investigated the contribution of hyperglycemia and the insulin/IGF axis. Immunohistochemistry identified placental cell types expressing MT1-MMP. MT1-MMP was compared between normal and GDM placentas by immunoblotting. Quantitative PCR of MT1-MMP in primary feto-placental endothelial cells (fpEC) and trophoblasts isolated from both normal and GDM placentas identified the cells contributing to the GDM-associated changes. A putative MT1-MMP role in angiogenesis was determined using blocking antibodies for in vitro angiogenesis assays. Potential GDM-associated factors and signaling pathways inducing MT1-MMP up-regulation in fpEC were identified using kinase inhibitors. Total and active MT1-MMP was increased in GDM placentas (+51 and 54%, respectively, Pfeto-placental endothelium, and insulin and IGF-II contribute. This may account for GDM-associated changes in the feto-placental vasculature.

  6. Effect of agonists and inhibitors of matrix metalloproteinases on expression of matrix metalloproteinase 1/2 and collagen Ⅰ in human scleral fibroblast%MMPs激动剂和抑制剂对人巩膜成纤维细胞MMP-1/2以及Ⅰ型胶原表达的作用研究

    Institute of Scientific and Technical Information of China (English)

    邹蕾蕾; 刘睿; 刘红; 黄莉雯; 周行涛; 周浩

    2013-01-01

    目的 研究基质金属蛋白酶(MMPs)家族MMP-1、MMP-2激动剂和抑制剂对人巩膜Ⅰ型胶原表达的作用,探讨MMP-1和MMP-2在人巩膜胶原代谢中的作用.方法 应用人巩膜成纤维细胞系作为研究模型,用荧光定量检测MMPs激动剂醋酸氨基苯汞(APMA)及抑制剂对人巩膜成纤维细胞MMP-1和MMP-2活性的影响;实时荧光定量反转录聚合酶链反应以及免疫印迹蛋白定量法检测比较激动剂组与抑制剂组对MMP-1/2以及Ⅰ型胶原mRNA和蛋白表达的变化;最后用siRNA干扰敲除MMP-1、MMP-2或MMP-1/2后,检测巩膜成纤维细胞Ⅰ型胶原蛋白水平的变化.结果 MMPs激动剂引起MMP-1/2活性显著增加,Ⅰ型胶原mRNA和蛋白表达下降,MMPs抑制剂引起MMP-1/2活性显著下降,相应的Ⅰ型胶原mRNA和蛋白表达增加(均P<O.01).激动剂可引起MMP-1 mRNA表达增加(P<0.01),对MMP-2 mRNA表达无明显影响(P>0.05),而抑制剂能引起MMP-2 mRNA表达下降(P<0.01),但对MMP-1 mRNA表达无显著影响(P>0.05).应用siRNA干扰方法敲除MMP-1、MMP-2或两者同时被敲除后,MMP-1蛋白表达均显著降低(均P<0.01),MMP-2除在MMP-1 siRNA干扰敲除组无明显变化外(P>0.05),其余各组均显著下降(均P<O.01);Ⅰ型胶原的表达在各敲除组中显著增加,当MMP-1和MMP-2均干扰敲除时,Ⅰ型胶原的表达增加最为显著(均P<0.01).结论 在人巩膜成纤维细胞系中,MMP-1/2激动剂和抑制剂均能引起MMP-1/2活性改变及相应的MMP-1/2蛋白表达变化,并最终导致Ⅰ型胶原表达的变化;MMP-1和MMP-2是调控人巩膜Ⅰ型胶原重塑的关键因子.%Objective To investigate the effect of agonist and inhibitor of matrix metalloproteinase-1 ( MMP-1) and MMP-2 in matrix metalloproteinases ( MMPs) on the expression of type I collagen and the effect of MMP-1 and MMP-2 in the development of human scleral collagen. Methods Human scleral fibroblast line was established as a model

  7. Matrix metalloproteinase 13 (MMP13) and tissue inhibitor of matrix metalloproteinase 1 (TIMP1), regulated by the MAPK pathway, are both necessary for Madin-Darby canine kidney tubulogenesis.

    Science.gov (United States)

    Hellman, Nathan E; Spector, June; Robinson, Jonathan; Zuo, Xiaofeng; Saunier, Sophie; Antignac, Corinne; Tobias, John W; Lipschutz, Joshua H

    2008-02-15

    A classic model of tubulogenesis utilizes Madin-Darby canine kidney (MDCK) cells. MDCK cells form monoclonal cysts in three-dimensional collagen and tubulate in response to hepatocyte growth factor, which activates multiple signaling pathways, including the mitogen-activated protein kinase (MAPK) pathway. It was shown previously that MAPK activation is necessary and sufficient to induce the first stage of tubulogenesis, the partial epithelial to mesenchymal transition (p-EMT), whereas matrix metalloproteinases (MMPs) are necessary for the second redifferentiation stage. To identify specific MMP genes, their regulators, tissue inhibitors of matrix metalloproteinases (TIMPs), and the molecular pathways by which they are activated, we used two distinct MAPK inhibitors and a technique we have termed subtraction pathway microarray analysis. Of the 19 MMPs and 3 TIMPs present on the Canine Genome 2.0 Array, MMP13 and TIMP1 were up-regulated 198- and 169-fold, respectively, via the MAPK pathway. This was confirmed by two-dimensional and three-dimensional real time PCR, as well as in MDCK cells inducible for the MAPK gene Raf. Knockdown of MMP13 using short hairpin RNA prevented progression past the initial phase of p-EMT. Knockdown of TIMP1 prevented normal cystogenesis, although the initial phase of p-EMT did occasionally occur. The MMP13 knockdown phenotype is likely because of decreased collagenase activity, whereas the TIMP1 knockdown phenotype appears due to increased apoptosis. These data suggest a model, which may also be important for development of other branched organs, whereby the MAPK pathway controls both MDCK p-EMT and redifferentiation, in part by activating MMP13 and TIMP1.

  8. QM/MM Studies of the Matrix Metalloproteinase 2 (MMP2) Inhibition Mechanism of (S)-SB-3CT and its Oxirane Analogue.

    Science.gov (United States)

    Zhou, Jia; Tao, Peng; Fisher, Jed F; Shi, Qicun; Mobashery, Shahriar; Schlegel, H Bernhard

    2010-11-09

    SB-3CT, (4-phenoxyphenylsulfonyl)methylthiirane, is a potent, mechanism-based inhibitor of the gelatinase sub-class of the matrix metalloproteinase (MMP) family of zinc proteases. The gelatinase MMPs are unusual in that there are several examples where both enantiomers of a racemic inhibitor have comparable inhibitory abilities. SB-3CT is one such example. Here, the inhibition mechanism of the MMP2 gelatinase by the (S)-SB-3CT enantiomer and its oxirane analogue is examined computationally, and compared to the mechanism of (R)-SB-3CT. Inhibition of MMP2 by (R)-SB-3CT was shown previously to involve enzyme-catalyzed C-H deprotonation adjacent to the sulfone, with concomitant opening by β-elimination of the sulfur of the three-membered thiirane ring. Similarly to the R enantiomer, (S)-SB-3CT was docked into the active site of MMP2, followed by molecular dynamics simulation to prepare the complex for combined quantum mechanics and molecular mechanics (QM/MM) calculations. QM/MM calculations with B3LYP/6-311+G(d,p) for the QM part (46 atoms) and the AMBER force field for the MM part were used to compare the reaction of (S)-SB-3CT and its oxirane analogue in the active site of MMP2 (9208 atoms). These calculations show that the barrier for the proton abstraction coupled ring opening reaction of (S)-SB-3CT in the MMP2 active site is 4.4 kcal/mol lower than its oxirane analogue, and the ring opening reaction energy of (S)-SB-3CT is only 1.6 kcal/mol less exothermic than its oxirane analogue. Calculations also show that the protonation of the ring-opened products by water is thermodynamically much more favorable for the alkoxide obtained from the oxirane, than for the thiolate obtained from the thiirane. In contrast to (R)-SB-3CT and the R-oxirane analogue, the double bonds of the ring-opened products of (S)-SB-3CT and its S-oxirane analogue have the cis-configuration. Vibrational frequency and intrinsic reaction path calculations on a reduced size QM/MM model (2747 atoms

  9. Preliminary evidence for a matrix metalloproteinase-2 (MMP-2)-dependent shedding of soluble CD40 ligand (sCD40L) from activated platelets.

    Science.gov (United States)

    Reinboldt, Stephan; Wenzel, Folker; Rauch, Bernhard H; Hohlfeld, Thomas; Grandoch, Maria; Fischer, Jens W; Weber, Artur-Aron

    2009-09-01

    Platelets are the major source of soluble CD40 ligand (sCD40L) in the blood. It has been demonstrated that CD40L is cleaved from the surface of activated platelets to release sCD40L. However, the enzyme involved in sCD40L shedding has not been identified yet. Using a panel of pharmacological inhibitors of serine, cysteine, aspartate, or metalloproteinases, preliminary evidence is presented for the hypothesis that matrix metalloproteinase-2 (MMP-2) might be the protease, primarily responsible for CD40L cleavage from platelet surface.

  10. Correlation of reversion-inducing cysteine-rich protein with kazal motifs (RECK) and extracellular matrix metalloproteinase inducer (EMMPRIN), with MMP-2, MMP-9, and survival in colorectal cancer.

    NARCIS (Netherlands)

    Jagt, M. van der; Sweep, C.G.J.; Waas, E.T.; Hendriks, T.; Ruers, T.J.M.; Merry, A.H.; Wobbes, Th.; Span, P.N.

    2006-01-01

    mRNA, and latent and active levels MMP-2 and -9 were higher in tumor tissue compared to normal tissue from 63 patients with colorectal cancer, whereas RECK and EMMPRIN levels were lower. Correlations between mRNA, latent, and active MMP were particular high for MMP-2 in tumor tissue (R(s)=0.6-0.8, P

  11. Relationship of Matrix Metalloproteinase-2 and-9 Expression to Gastric Carcinoma Invasion and Metastasis%MMP-2、MMP-9与胃癌侵袭、转移的关系

    Institute of Scientific and Technical Information of China (English)

    钟华; 廖爱军; 刘迪群

    2012-01-01

    目的 探讨MMP-2、MMP-9的表达与胃癌侵袭和转移的关系.方法 采用免疫组化方法检测44例胃癌患者胃黏膜标本中MMP-2、MMP-9的表达情况.结果 有浆膜浸润的胃癌患者其MMP-2、MMP-9的阳性表达率均显著高于无浆膜浸润的胃癌患者(均P<0.01),有淋巴结转移的胃癌患者其MMP-2、MMP-9的阳性表达率均显著高于无淋巴结转移的胃癌患者(均P<0.01).结论 MMP-2、MMP-9表达与胃癌侵袭、转移有关,其表达增加提示胃癌有较高的恶性生物学行为.MMP-2、MMP-9可能成为评价胃癌恶性生物学特性的指标.

  12. 子宫颈鳞癌中MMP-2,MMP-9的表达及意义%Expression of Matrix Metalloproteinase in Cervical Squamous Cell Carcinoma and Its Significance

    Institute of Scientific and Technical Information of China (English)

    林蓓; 蔡威; 张淑兰

    2003-01-01

    目的: 探讨基质金属蛋白酶(MMP-2,MMP-9)与宫颈鳞癌的发生、发展及侵袭的关系.方法: 32例宫颈鳞状细胞癌为实验组,慢性宫颈炎及正常宫颈组织各30例为对照组.明胶酶谱法测定活性型MMP-2及MMP-9蛋白的含量,RT-PCR技术检测MMP-2及MMP-9 mRNA表达水平.结果:宫颈鳞癌组织中MMP-2及MMP-9酶活性和MMP-2及MMP-9 mRNA表达量均显著高于慢性宫颈炎及正常宫颈组织(P均<0.01).结论: MMP-2及MMP-9与宫颈鳞癌的发生、发展及侵袭有关.

  13. THE ASSOCIATION BETWEEN MATRIX METALLOPROTEINASE-9 (MMP-9 WITH HIGH SENSITIVE TROPONIN T (hs-TnT IN PATIENT WITH ACUTE MYOCARDIAL INFARCTION

    Directory of Open Access Journals (Sweden)

    I Putu Gede Eka Ariawan Suyasa

    2015-10-01

    Full Text Available Mechanism of acute myocardial infarction (AMI is previously due to atherosclerotic plaque rupturewith occurs because extra-cellular matrix of plaque fibrous cap destruction or degradation by proteaseenzyme matrix metalloproteinase-9  (MMP-9, which released by macrophage cell.Increased plasmaMMP-9  is  predisposition  factor  of  atherosclerotic  plaque  rupture  in AMI  and  followed  by  acutethrombosis inside coronary artery lumen which caused myocardial ischemic and clinical sign of AMI. Ifthe ischemic process continuous and ongoing that can caused myocardial necrosis which can increasedplasma troponin. High sensitive troponin T (hs-TnT newly and more sensitive detection of plasmacTn-T  than conventional.The aim of  this study was  to determined  the association between plasmaMMP-9 with hs-TnT in AMI patients.This study was a cross-sectional observational which performedin 62 patients with AMI which enrolled by consecutive sampling at Sanglah Hospital Denpasar fromDecember 2011 until December 2012. MMP-9 and hs-TnT plasma level were measured 48 hours afteronset IMA. Sixty two patients AMI were involved in this study consist of 35 STEMI patients (56.5%and 27 NSTEMI patients (43.5%, the mean plasma MMP-9 was 23.9 (SD 0.42 ng/mL and hs-TnT was464.7 (SD 39.3 ng/mL.The results of this study were positive correlation between MMP-9 and hs-TnTAMI  patients  (r=  0.507; Y=  -  650.6 +  46.7(X1; P<0.0001;  plasma MMP-9  and  onset  of AMI wereinfluenced to plasma hs-TnT with formulationY = - 815.0 + 46.5(X1+ 20.7(X2; (â MMP-9=46.5(95%CI: 24.7 to 68.4; P<0.0001; â onset AMI=20.7(95%CI : 2.1 to 39.4; P=0.030 and there was more strongercorrelation betweenMMP-9 and hs-TnT in STEMI group than NSTEMI. [MEDICINA 2015;46:22-27].Mekanisme terjadinya infark miokard akut (IMA didahului oleh proses ruptur plak aterosklerosisdan diawali dengan destruksi atau degradasi matriks ekstraseluler fibrus cap plak oleh enzim proteaseyang

  14. Minimally invasive colorectal resection is associated with significantly elevated levels of plasma matrix metalloproteinase 3 (MMP-3) during the first month after surgery which may promote the growth of residual metastases.

    Science.gov (United States)

    Shantha Kumara, H M C; Gaita, David J; Miyagaki, Hiromichi; Yan, Xiaohong; Herath, Sonali A C; Cekic, Vesna; Whelan, Richard L

    2014-12-01

    MMP-3, a member of the matrix metalloproteinase (MMP) family, is involved in the breakdown of the extracellular matrix in tissue remodeling and may also play a role in cancer progression and metastasis. Minimally invasive colorectal resection (MICR) may increase plasma MMP-3 levels directly via surgical trauma or indirectly due to surgery-associated elevations in TNF-α and IL1 which are regulators of MMP-3. This study's purpose was to evaluate plasma MMP-3 levels during the first month after MICR for colorectal cancer. Patients enrolled in an IRB approved data/plasma bank who underwent elective MICR for CRC. Blood plasma samples had been collected preoperatively, on postoperative day (POD) 1, 3 and at varying postoperative time points and were stored at -80 °C. The late samples (POD 7-41) were bundled into 7 day time blocks and considered as single time points. MMP-3 levels were analyzed in duplicate via ELISA and the results reported as mean ± SD. The paired t test was used for analysis (significance, p MICR met the inclusion criteria. The mean PreOp MMP-3 level was 14.9 ± 7.8 ng/ml (n = 73). Significantly elevated mean plasma levels were noted on POD 1 (21.4 ± 14.7 ng/ml, n = 73, p MICR for CRC. The early postoperative increase in MMP-3 levels may be due to the surgery-related acute inflammatory response; the elevation noted during weeks 2-3 may be related to wound healing. Increased MMP-3 levels may promote metastases or the growth of residual cancer.

  15. Correlative studies of the expression of matrix metalloproteinase 2 and 9 in human bladder cancer%MMP-2和MMP-9在膀胱癌组织中的表达及相关性研究

    Institute of Scientific and Technical Information of China (English)

    陈国俊; 王健

    2007-01-01

    目的:研究MMP-2和MMP-9在膀胱移行细胞癌组织中的表达及其与膀胱癌侵袭、转移的关系.方法:应用链霉菌抗生物素蛋白-过氧化酶(S-P)免疫组化方法,检测5例正常膀胱组织和49例膀胱移行细胞癌组织中MMP-2和MMP-9的表达水平.结果:MMP-2、MMP-9在膀胱正常组织中均呈阴性表达,在49例膀胱癌组织中均不同程度的表达.随肿瘤分级的增高,MMP-2和MMP-9的表达显著增强(P值均小于0.05);随肿瘤分期的增高,MMP-2的表达也显著增强(P<0.05),而MMP-9在膀胱肿瘤低期和高期之间无显著性差异(P>0.05).结论:MMP-2、MMP-9与膀胱癌的侵袭转移有关,且MMP-2可能对判断膀胱癌的预后有帮助.

  16. Discovery of novel, highly potent, and selective quinazoline-2-carboxamide-based matrix metalloproteinase (MMP)-13 inhibitors without a zinc binding group using a structure-based design approach.

    Science.gov (United States)

    Nara, Hiroshi; Sato, Kenjiro; Naito, Takako; Mototani, Hideyuki; Oki, Hideyuki; Yamamoto, Yoshio; Kuno, Haruhiko; Santou, Takashi; Kanzaki, Naoyuki; Terauchi, Jun; Uchikawa, Osamu; Kori, Masakuni

    2014-11-13

    Matrix metalloproteinase-13 (MMP-13) has been implicated to play a key role in the pathology of osteoarthritis. On the basis of X-ray crystallography, we designed a series of potent MMP-13 selective inhibitors optimized to occupy the distinct deep S1' pocket including an adjacent branch. Among them, carboxylic acid inhibitor 21k exhibited excellent potency and selectivity for MMP-13 over other MMPs. An effort to convert compound 21k to the mono sodium salt 38 was promising in all animal species studied. Moreover, no overt toxicity was observed in a preliminary repeat dose oral toxicity study of compound 21k in rats. A single oral dose of compound 38 significantly reduced degradation products (CTX-II) released from articular cartilage into the joint cavity in a rat MIA model in vivo. In this article, we report the discovery of highly potent, selective, and orally bioavailable MMP-13 inhibitors as well as their detailed structure-activity data.

  17. The interferon-gamma-induced GTPase, mGBP-2, inhibits tumor necrosis factor alpha (TNF-alpha) induction of matrix metalloproteinase-9 (MMP-9) by inhibiting NF-kappaB and Rac protein.

    Science.gov (United States)

    Balasubramanian, Sujata; Fan, Meiyun; Messmer-Blust, Angela F; Yang, Chuan H; Trendel, Jill A; Jeyaratnam, Jonathan A; Pfeffer, Lawrence M; Vestal, Deborah J

    2011-06-03

    Matrix metalloproteinase-9 (MMP-9) is important in numerous normal and pathological processes, including the angiogenic switch during tumor development and tumor metastasis. Whereas TNF-α and other cytokines up-regulate MMP-9 expression, interferons (IFNs) inhibit MMP-9 expression. We found that IFN-γ treatment or forced expression of the IFN-induced GTPase, mGBP-2, inhibit TNF-α-induced MMP-9 expression in NIH 3T3 fibroblasts, by inhibiting MMP-9 transcription. The NF-κB transcription factor is required for full induction of MMP-9 by TNF-α. Both IFN-γ and mGBP-2 inhibit the transcription of a NF-κB-dependent reporter construct, suggesting that mGBP-2 inhibits MMP-9 induction via inhibition of NF-κB-mediated transcription. Interestingly, mGBP-2 does not inhibit TNF-α-induced degradation of IκBα or p65/RelA translocation into the nucleus. However, mGBP-2 inhibits p65 binding to a κB oligonucleotide probe in gel shift assays and to the MMP-9 promoter in chromatin immunoprecipitation assays. In addition, TNF-α activation of NF-κB in NIH 3T3 cells is dependent on Rac activation, as evidenced by the inhibition of TNF-α induction of NF-κB-mediated transcription by a dominant inhibitory form of Rac1. A role for Rac in the inhibitory action of mGBP-2 on NF-κB is further shown by the findings that mGBP-2 inhibits TNF-α activation of endogenous Rac and constitutively activate Rac can restore NF-κB transcription in the presence of mGBP-2. This is a novel mechanism by which IFNs can inhibit the cytokine induction of MMP-9 expression.

  18. 转录因子Ets-2调控小鼠成釉细胞MMP-20基因表达的研究%Study of Transcription Factor Ets-2 on Regulating the Expression of Matrix Metalloproteinase-20

    Institute of Scientific and Technical Information of China (English)

    孙霞; 王青山; 韩婷婷; 李东亮; 孙学玲; 孙岩; 纪虹利; 高玉光

    2013-01-01

    Objective: To study the significance of Ets - 2 in developing dental enamel. Methods: Immunocyto-chemistry technique were used to observe and analyze the expression of Ets - 2 gene in incisors of postnatal 5 d mouse. The 200ng pcDNA3. 1/myc- HisA-Ets-2 and the siRNA for Ets -2 mRNA were transfected ALC respectively. Then quantitative real -time PCR was used to measure the different expression of MMP-20. Dual lu-ciferase analysis was used to observe the effects of Ets-2 on the transcriptional activity of MMP20 promoter after mutation of the Ets-2 binding sites. Results: Ets -2 was detected in amelogenesis by immunocytochemistry. By real time PCR, the up-regulation of Ets-2 directly result in a significant higher expression of MMP-20. Contrary to it, knockdown of the transcription factor Ets-2 by siRNA inhibited the MMP20 expression. By the Dual -Luciferase Reporter Assay System, compared with the control group , the transcriptional activity of MMP20 which was transfected with Ets - 2 was raised. After mutation of the Ets-2 binding sites, the transcriptional activity of MMP20 had a marked decline. Ets -2 lost the use of regulating the transcriptional activity of MMP-20. Conclusion: Ets-2 up-regulates MMP-20 gene expression via the combination with the Ets - 2 binding sites of MMP-20 in ameloblasts.%目的:通过研究Ets-2转录因子对调控小鼠成釉细胞金属基质蛋白酶-20(matrix metalloproteinase-20,MMP-20)基因表达的调控作用,进一步明确Ets-2在釉质发育中的作用.方法:应用免疫组化方法观察出生后5d小鼠切牙成釉细胞中Ets-2的表达;在小鼠成釉细胞中分别转染200ng pcDNA3.1/myc-HisA-Ets-2和Ets-2 siRNA后,利用实时定量RT-PCR法检测MMP20基因表达的不同变化;用双荧光素酶报告基因检测系统分析Ets-2对MMP-20启动子突变后转录活性的影响.结果:免疫组化显示Ets-2在成釉细胞中呈阳性表达.实时定量RT-PCR研究发现Ets-2过表达后,MMP-20表

  19. Expression of Matrix Metalloproteinase-2 and -9 in Carcinoma of Esophagus%食管癌基质金属蛋白酶MMP-2和MMP-9表达的意义

    Institute of Scientific and Technical Information of China (English)

    王雪梅

    2005-01-01

    目的:探讨基质金属蛋白酶MMP-2和MMP-9在食管癌组织中的表达及其与食管癌浸润转移的关系.方法:用免疫组化S-P法分别检测59例食管癌组织中MMP-2和MMP-9的表达.结果:MMP-2和MMP-9阳性表达率食管腺癌分别为84.61%、76.92%,明显高于食管鳞癌50.00%、43.48%,P<0.05;淋巴结转移组分别为93.33%、86.67%,明显高于无淋巴结转移组41.38%、37.93%,P<0.01.结论:MMP-2和MMP-9在食管癌侵袭及转移过程中可能发挥重要作用.

  20. Extracellular S100A4(mts1) stimulates invasive growth of mouse endothelial cells and modulates MMP-13 matrix metalloproteinase activity

    DEFF Research Database (Denmark)

    Schmidt-Hansen, Birgitte; Ornås, Dorte; Grigorian, Mariam

    2004-01-01

    S100A4(mts1) protein expression has been strongly associated with metastatic tumor progression. It has been suggested as a prognostic marker for a number of human cancers. It is proposed that extracellular S100A4 accelerates cancer progression by stimulating the motility of endothelial cells......, thereby promoting angiogenesis. Here we show that in 3D culture mouse endothelial cells (SVEC 4-10) respond to recombinant S100A4 by stimulating invasive growth of capillary-like structures. The outgrowth is not dependent on the stimulation of cell proliferation, but rather correlates...... with the transcriptional modulation of genes involved in the proteolytic degradation of extracellular matrix (ECM). Treatment of SVEC 4-10 with the S100A4 protein leads to the transcriptional activation of collagenase 3 (MMP-13) mRNA followed by subsequent release of the protein from the cells. Beta-casein zymography...

  1. 胃肠道间质瘤组织中MMP-13与CD147的表达及其意义%EXPRESSIONS OF CD147 AND MATRIX METALLOPROTEINASE-13 IN GASTROINTESTINAL STROMAL TUMORS AND THEIR SIGNIFICANCE

    Institute of Scientific and Technical Information of China (English)

    张薇; 潘献柱; 吴正升; 吴强

    2009-01-01

    目的 探讨基质金属蛋白酶-13(Matrix metalloproteinases-13,MMP-13)和CD147的表达与胃肠道间质瘤(Gastrointestinals stromal tumors,GISTs)临床病理特征的关系.方法 采用免疫组织化学检测69例GISTs组织中MMP13及CD147的表达,分析MMP-13和CD147与各病理参数的关系.结果 GISTs组织中MMP-13和CD147的表达率分别为95.7%和97.1%,MMP-13阳性表达程度与GISTs的生物学行为、CD147表达及肿瘤的大小呈正相关(P<0.05);CD147阳性表达程度与肿瘤的大小呈正相关(P<0.05).结论 MMP-13表达与GISTs生物学行为关系密切,可作为GISTs生物学行为的潜在评价指标.

  2. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette;

    2010-01-01

    To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1) in cancer...

  3. SNAP23, Syntaxin4, and vesicle-associated membrane protein 7 (VAMP7) mediate trafficking of membrane type 1-matrix metalloproteinase (MT1-MMP) during invadopodium formation and tumor cell invasion.

    Science.gov (United States)

    Williams, Karla C; McNeilly, Rachael E; Coppolino, Marc G

    2014-07-01

    Movement through the extracellular matrix (ECM) requires cells to degrade ECM components, primarily through the action of matrix metalloproteinases (MMPs). Membrane type 1-matrix metalloproteinase (MT1-MMP) has an essential role in matrix degradation and cell invasion and localizes to subcellular degradative structures termed invadopodia. Trafficking of MT1-MMP to invadopodia is required for the function of these structures, and here we examine the role of N-ethylmaleimide-sensitive factor-activating protein receptor (SNARE)-mediated membrane traffic in the transport of MT1-MMP to invadopodia. During invadopodium formation in MDA-MB-231 human breast cancer cells, increased association of SNAP23, Syntaxin4, and vesicle-associated membrane protein 7 (VAMP7) is detected by coimmunoprecipitation. Blocking the function of these SNAREs perturbs invadopodium-based ECM degradation and cell invasion. Increased level of SNAP23-Syntaxin4-VAMP7 interaction correlates with decreased Syntaxin4 phosphorylation. These results reveal an important role for SNARE-regulated trafficking of MT1-MMP to invadopodia during cellular invasion of ECM.

  4. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette

    2010-01-01

    To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1) in cancer...

  5. Expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinases 1 (TIMP-1) by colorectal cancer cells and adjacent stroma cells--associations with histopathology and patients outcome

    DEFF Research Database (Denmark)

    Jensen, Søren Astrup; Vainer, Ben; Bartels, Annette

    2010-01-01

    AIM: To elucidate cellular features accountable for colorectal cancers' (CRC) capability to invade normal tissue and to metastasize, we investigated the level of the collagenase matrix metalloproteinase 9 (MMP-9) and its physiological inhibitor tissue inhibitor of metalloproteinases 1 (TIMP-1...

  6. Expression and effect of matrix metalloproteinase 2 in human fetal scleral fibroblasts treated with an extremely low frequency electromagnetic field%极低频电磁辐射下HFSF中MMP-2的表达及作用

    Institute of Scientific and Technical Information of China (English)

    田甜; 朱煌

    2015-01-01

    Objective To observe the matrix metalloproteinase 2 (MMP-2) activity and protein expression in human fetal scleral fibroblasts (HFSFs) exposed to an extremely low frequency electromagnetic field (ELF-EMF) and its impact on collagen Ⅰ synthesis.Methods This was an experimental study.HFSFs were cultured and divided into a control group,radiation group and inhibitor group according to their exposure to 50 Hz,0.2 mT electromagnetic field or joint MMP-2 specific inhibitors (sc-204092).MMP-2 activity was measured by gelatin enzymography.MMP-2 and collagen Ⅰ protein expression were detected by Western Blot.A single factor analysis of variance was used to determine whether there was a difference among these groups.Results There were significant differences between the three groups (F=14.96,139.88,63.46,P<0.01).Compared to the control group,MMP-2 activity and protein expression increased in the radiation group (P<0.05) and collagen Ⅰ protein expression decreased (P<0.01).Compared to the radiation group,MMP-2 activity and protein expression decreased in the inhibitor group (P<0.01) and collagen Ⅰ protein expression increased (P<0.05).Conclusion ELF-EMF may regulate the synthesis of collagen Ⅰ by affecting the activity and protein expression of MMP-2.%目的 研究极低频电磁场(ELF-EMF)作用下人胚胎眼巩膜成纤维细胞(HFSF)中基质金属蛋白酶-2(MMP-2)活性与蛋白表达的变化以及ELF-EMF对Ⅰ型胶原(Collagen Ⅰ)合成的影响.方法 实验研究.根据是否暴露于50 Hz、0.2 mT电磁场或加入MMP-2特异性抑制剂(sc-204092),将实验细胞分为对照组、辐照组和抑制剂组.明胶酶谱法检测各组HFSF中MMP-2酶活性的变化;Western Blot法检测各组HFSF中MMP-2、Collagen Ⅰ蛋白水平的变化.采用单因素方差分析进行数据分析.结果 3组HFSF细胞中MMP-2酶活性、MMP-2、Collagen Ⅰ蛋白表达比较差异均具有统计学意义(F=14.96、139.88、63.46,P均<0.01);辐照组较对照组MMP

  7. Plasma MMP1 and MMP8 expression in breast cancer: Protective role of MMP8 against lymph node metastasis

    Directory of Open Access Journals (Sweden)

    Christiaens Marie-Rose

    2008-03-01

    Full Text Available Abstract Background Elevated levels of matrix metalloproteinases have been found to associate with poor prognosis in various carcinomas. This study aimed at evaluating plasma levels of MMP1, MMP8 and MMP13 as diagnostic and prognostic markers of breast cancer. Methods A total of 208 breast cancer patients, of which 21 with inflammatory breast cancer, and 42 healthy controls were included. Plasma MMP1, MMP8 and MMP13 levels were measured using ELISA and correlated with clinicopathological characteristics. Results Median plasma MMP1 levels were higher in controls than in breast cancer patients (3.45 vs. 2.01 ng/ml, while no difference was found for MMP8 (10.74 vs. 10.49 ng/ml. ROC analysis for MMP1 revealed an AUC of 0.67, sensitivity of 80% and specificity of 24% at a cut-off value of 4.24 ng/ml. Plasma MMP13 expression could not be detected. No correlation was found between MMP1 and MMP8 levels. We found a trend of lower MMP1 levels with increasing tumour size (p = 0.07; and higher MMP8 levels with premenopausal status (p = 0.06 and NPI (p = 0.04. The median plasma MMP1 (p = 0.02 and MMP8 (p = 0.007 levels in the non-inflammatory breast cancer patients were almost twice as high as those found in the inflammatory breast cancer patients. Intriguingly, plasma MMP8 levels were positively associated with lymph node involvement but showed a negative correlation with the risk of distant metastasis. Both controls and lymph node negative patients (pN0 had lower MMP8 levels than patients with moderate lymph node involvement (pN1, pN2 (p = 0.001; and showed a trend for higher MMP8 levels compared to patients with extensive lymph node involvement (pN3 and a strong predisposition to distant metastasis (p = 0.11. Based on the hypothesis that blood and tissue protein levels are in reverse association, these results suggest that MMP8 in the tumour may have a protective effect against lymph node metastasis. Conclusion In summary, we observed differences in MMP1

  8. 胃肠间质瘤组织中MMP-14与CD147的表达及其意义%Expression of CD147 and matrix metalloproteinases-14 in gastrointestinal stromal tumors and their significance

    Institute of Scientific and Technical Information of China (English)

    潘献柱; 吴强

    2013-01-01

    Objective To investigate the relationship between the expression of matrix metalloproteinases-14 and clinical significance in gastrointestinal stromal tumors (GISTs).Methods Sixty-nine cases of GISTs were immunohistochemically studied for the expression of MMP-14 and CD147,and their relations with every pathologic parameters were analyzed statistically.Results Out of 69 cases,the positivity of MMP-14 and CD147 was 94.2% and 97.1%,respectively.The expression levels of MMP-14 were significantly related to risk of GISTs,CD147 expression and size of tumors (P <0.05) ; The expression levels of CD147 were significantly related to size of tumors (P < 0.05).Conclusion MMP-14 is closely related with risk of GISTs,and therefore it might be a potential parameter for risk evaluation of GISTs.%目的 探讨基质金属蛋白酶-14(MMP-14)和CD147的表达与胃肠间质瘤(GISTs)临床病理特征的关系.方法 采用免疫组化SP法检测69例GISTs组织中MMP-14和CD147的表达,分析MMP-14和CD147与各病理参数的关系.结果 GISTs组织中MMP-14和CD147的阳性率分别为94.2%和97.1%,MMP-14阳性表达程度与GISTs的危险度、CD147表达及肿瘤的大小呈正相关(P<0.05);CD147阳性表达程度与肿瘤的大小呈正相关(P<0.05).结论 MMP-14阳性表达与GISTs危险度关系密切,可作为GISTs危险度的潜在评价指标.

  9. MMP3 — EDRN Public Portal

    Science.gov (United States)

    Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. MMP3 degrades fibronectin, laminin, collagens III, IV, IX, and X, and cartilage proteoglycans. The enzyme is thought to be involved in wound repair, progression of atherosclerosis, and tumor initiation. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.

  10. A monoclonal antibody interferes with TIMP-2 binding and incapacitates the MMP-2-activating function of multifunctional, pro-tumorigenic MMP-14/MT1-MMP

    DEFF Research Database (Denmark)

    Shiryaev, S A; Remacle, A G; Golubkov, V S;

    2013-01-01

    Matrix metalloproteinases (MMPs) and, especially membrane type 1 (MT1)-MMP/MMP-14, are promising drug targets in malignancies. In contrast with multiple small-molecule and protein pan-inhibitors of MT1-MMP cleavage activity, the murine 9E8 monoclonal antibody targets the MMP-2-activating function...... tissue inhibitor of metalloproteinases-2 (TIMP-2) association with MT1-MMP. As a result, the 9E8 antibody incapacitates the TIMP-2-dependent MMP-2-activating function alone rather than the general enzymatic activity of human MT1-MMP. The specific function of the 9E8 antibody we determined directly...... supports an essential, albeit paradoxical, role of the protein inhibitor (TIMP-2) in MMP-2 activation via a unique membrane-tethered mechanism. In this mechanism, the formation of a tri-molecular MT1-MMPTIMP-2MMP-2 complex is required for both the capture of the soluble MMP-2 proenzyme by cells...

  11. Matrix metalloproteinase-2 (MMP-2) and its tissue inhibitor (TIMP-2) are prognostic factors in cervical cancer, related to invasive disease but not to high-risk human papillomavirus (HPV) or virus persistence after treatment of CIN.

    Science.gov (United States)

    Branca, M; Ciotti, M; Giorgi, C; Santini, D; Di Bonito, L; Costa, S; Benedetto, A; Bonifacio, D; Di Bonito, P; Paba, P; Accardi, L; Syrjänen, S; Favalli, C; Syrjänen, K

    2006-01-01

    Matrix metalloproteinase-2 (MMP-2) and its tissue inhibitor (TIMP-2) are important regulators of cancer invasion and metastasis. Their associations to high-risk (HR) human papillomavirus (HPV) in cervical intra-epithelial neoplasia (CIN) and cervical cancer (CC) are unexplored and their prognostic significance in CC remains controversial. As part of our HPV-PathogenISS study, a series of 150 CCs and 152 CIN lesions were examined using immunohistochemical (IHC) staining for MMP-2 and TIMP-2 and tested for HPV using PCR with 3 primer sets (MY09/11, GP5+/GP6+, SPF). Follow-up data were available from all squamous cell carcinoma patients and 67 CIN lesions had been monitored with serial PCR for HPV after cone treatment. MMP-2 increased with the grade of CIN, with major up-regulation upon transition to invasive cancer (OR 20.78) (95%CI 7.16-60.23) (p=0.0001). TIMP-2 retained its normal expression until CIN3, with dramatic down-regulation in invasive disease (p=0.0001 for trend). Thus, the MMP2:TIMP-2 ratio increased with progressive CIN, exceeding the value 1.0 only in invasive disease. Both MMP-2 and TIMP-2 are highly specific (TIMP-2; 100%) discriminators of CIN with 100% positive predictive value (TIMP-2), but suffer from low sensitivity and negative predictive value. Neither MMP-2 nor TIMP-2 showed any significant association with HR HPV or virus persistence/clearance. TIMP-2 (but not MMP-2) was a significant predictor of survival in univariate (Kaplan-Meier) analysis (p=0.007), but lost its significance in multivariate (Cox) analysis. The activities of MMP-2 and TIMP-2 in cervical carcinogenesis seem to be unrelated to HR-HPV The inverse MMP-2:TIMP-2 ratio is a sign of poor prognosis. A combination of a TIMP-2 assay with another test showing high SE and high NPV (e.g., HCII for HPV) should provide a potential screening tool capable of accurate detection of CIN.

  12. Hematopoietic Stem Cell Mobilization and Homing after Transplantation: The Role of MMP-2, MMP-9, and MT1-MMP

    Directory of Open Access Journals (Sweden)

    Neeta Shirvaikar

    2012-01-01

    Full Text Available Hematopoietic stem/progenitor cells (HSPCs are used in clinical transplantation to restore hematopoietic function. Here we review the role of the soluble matrix metalloproteinases MMP-2 and MMP-9, and membrane type (MT1-MMP in modulating processes critical to successful transplantation of HSPC, such as mobilization and homing. Growth factors and cytokines which are employed as mobilizing agents upregulate MMP-2 and MMP-9. Recently we demonstrated that MT1-MMP enhances HSPC migration across reconstituted basement membrane, activates proMMP-2, and contributes to a highly proteolytic bone marrow microenvironment that facilitates egress of HSPC. On the other hand, we reported that molecules secreted during HSPC mobilization and collection, such as hyaluronic acid and thrombin, increase MT1-MMP expression in cord blood HSPC and enhance (prime their homing-related responses. We suggest that modulation of MMP-2, MMP-9, and MT1-MMP expression has potential for development of new therapies for more efficient mobilization, homing, and engraftment of HSPC, which could lead to improved transplantation outcomes.

  13. 姜黄素对心肌梗死后血流动力学及心肌基质金属蛋白酶-2表达的影响%Effects of curcumin on MMP-2 and hemodynamics following myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    贺兆发; 刘春辉; 刘聪辉; 卢均坤; 李奕红; 范蕾; 刘畅; 张丽

    2015-01-01

    Objective The present study was designed to examine the effects of curcumin on hemodynamics and ma-trix metallo proteinases-2 ( MMP-2 ) expression in myocardial infarction rats. Method 70 cases Sprague-Dawley healthy adult male rats (250~300g) were randomly divided into sham operation group (n=15) and miocardial in-farction group ( MI, n=55 ) . The mode of operation was that the line in left anterior descending coronary artery threading and knotting but not to block the blood flow had used for sham operation group. MI group was established by ligation of left anterior descending branch of coronary artery and the sham operation group ( sham) rats underwent the same procedure without ligation. 24 hours later, the MI group was further sub-divided into 3 groups: control group, solvent group and curcumin group. The solvent group was treated with only solvent( a blend which blend Po-lyethylene glycol, alcohol and water)and curcumin group was intraperitoneally administered with curumin at 100mg/kg/d for 28 days. MP150 type multichannel physiological functions signal collection processing system was used to test left ventricular end diastolic pressure ( LVDEP) , systolic pressure ( SYS) , maximum change in pressure over the cycle (dp/dtmax), minimum change in pressure over the cycle (dp/dtmin) and mean blood pressure (MBP). HE stalning was employed to detect the morphological changes of cardiomyocytes. Moreover, the expression of MMP-2 protein in the myocardium of the rats was tested by immunohistochemical technique. Result SYS, dp/dtmax and dp/dtmin were significantly increased while the LVDEP decreased in the curcumin group compared with control (P<0. 05). Immunohistochemistry revealed a decrease in expression of MMP2 in curcumin group compared to con-tral and solvent groups(P<0. 05). HE stalning revealed that some normal cardiac myocytes were disappeared after MI, and were replaced by collagens. Conclusion Our study revealed that curcumin ameliorated hemodynamics and

  14. 9-Hydroxypheophorbide α-mediated photodynamic therapy induces matrix metalloproteinase-2 (MMP-2) and MMP-9 down-regulation in Hep-2 cells via ROS-mediated suppression of the ERK pathway.

    Science.gov (United States)

    Zhang, Huankang; Shen, Bo; Swinarska, Joanna T; Li, Wen; Xiao, Kuanlin; He, Peijie

    2014-03-01

    Photodynamic therapy (PDT) is a promising treatment modality for malignant diseases through the generation of reactive oxygen species (ROS). In this study, we assessed the change of migration and invasion of HEp-2 cells after sublethal doses of 9-hydroxypheophorbide α (9-HPbD)-mediated PDT in vitro, and explored the role of ROS in 9-HPbD-PDT-induced anti-metastatic effects in HEp-2 cells. Following PDT, ROS were measured by a fluorescence microscope in both the presence and absence of glutathione (GSH) pretreatment. Wound healing assay, cell migration assay, and matrigel invasion assay were used to evaluate the cellular migration and invasion. Western blot was performed to investigate the signaling pathways that may have been involved. ROS were rapidly generated in 9-HPbD-loaded HEp-2 laryngeal cancer cells by the activation of a diode laser and were significantly inhibited by a 6-h GSH pretreatment. Wound healing assay, cell migration assay, and matrigel invasion assay showed that sublethal PDT significantly suppressed the migration and invasion of HEp-2 cells. GSH decreased the ability of PDT to inhibit the invasion of HEp-2 cells. Western blot analysis showed that PDT significantly inhibited the phosphorylation of MEK1/2 and ERK1/2, and significantly suppressed the expression of MMP-2 and MMP-9 after 24h following the implementation of sublethal PDT, and these efficacies of PDT could be abrogated by GSH pretreatment. 9-HPbD-PDT attenuated the migration and invasion of HEp-2 cells in vitro, which may be related to the down-regulated expression of MMP-2 and MMP-9 via ROS-mediated-inhibition of phosphorylation in the ERK/MEK signaling pathway. Copyright © 2014. Published by Elsevier B.V.

  15. Structural differences of matrix metalloproteinases with potential implications for inhibitor selectivity examined by the GRID/CPCA approach

    DEFF Research Database (Denmark)

    Terp, Gitte Elgaard; Cruciani, Gabriele; Christensen, Inge Thøger

    2002-01-01

    The matrix metalloproteinases (MMPs) are a family of proteolytic enzymes, which have been the focus of a lot of research in recent years because of their involvement in various disease conditions. In this study, structures of 10 enzymes (MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP12, MMP13, MMP14...

  16. Tumour-targeting properties of antibodies specific to MMP-1A, MMP-2 and MMP-3

    Energy Technology Data Exchange (ETDEWEB)

    Pfaffen, Stefanie; Frey, Katharina; Stutz, Irene; Roesli, Christoph; Neri, Dario [Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zuerich, Zuerich (Switzerland)

    2010-08-15

    Matrix metalloproteinases (MMPs), a group of more than 20 zinc-containing endopeptidases, are upregulated in many diseases, but several attempts to use radiolabelled MMP inhibitors for imaging tumours have proved unsuccessful in mouse models, possibly due to the limited specificity of these agents or their unfavourable pharmacokinetic profiles. In principle, radiolabelled monoclonal antibodies could be considered for the selective targeting and imaging of individual MMPs. We cloned, produced and characterized high-affinity monoclonal antibodies specific to murine MMP-1A, MMP-2 and MMP-3 in SIP (small immunoprotein) miniantibody format using biochemical and immunochemical methods. We also performed comparative biodistribution analysis of their tumour-targeting properties at three time points (3 h, 24 h, 48 h) in mice bearing subcutaneous F9 tumours using radioiodinated protein preparations. The clinical stage L19 antibody, specific to the alternatively spliced EDB domain of fibronectin, was used as reference tumour-targeting agent for in vivo studies. All anti-MMP antibodies and SIP(L19) strongly stained sections of F9 tumours when assessed by immunofluorescence methods. In biodistribution experiments, SIP(SP3), specific to MMP-3, selectively accumulated at the tumour site 24 and 48 h after intravenous injection, but was rapidly cleared from other organs. By contrast, SIP(SP1) and SIP(SP2), specific to MMP-1A and MMP-2, showed no preferential accumulation at the tumour site. Antibodies specific to MMP-3 may serve as vehicles for the efficient and selective delivery of imaging agents or therapeutic molecules to sites of disease. (orig.)

  17. 间接性TMJ损伤后髁突软骨中MMP-13表达及其意义%EXPRESSION OF MATRIX METALLOPROTEINASE- 13 IN THE CONDYLE OF TEMPOROMANDIBULAR JOINT AFTER INDIRECT TRAUMA

    Institute of Scientific and Technical Information of China (English)

    李新军; 胡开进; 王文勇; 李玉松; 周树夏; 孔亮

    2003-01-01

    目的通过观察羊颞下颌关节(temporomandibularjoint,TMJ)间接创伤后不同时期髁突软骨中基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)的表达,探讨MMP-13在TMJ创伤后导致颞下颌关节骨关节病(temporomandibularjoint osteoarthrosis,TMJOA)过程中的作用.方法用自制撞击装置造成山羊双侧颞颌关节间接性创伤.分别于伤后1周、1个月、3个月、6个月取材,并以正常TMJ作为对照,用免疫荧光组织化学方法进行观察.结果TMJ髁突软骨中MMP-13伤后1周即出现强的表达,以后逐渐减弱;荧光着色于软骨破坏区增生滑膜样组织及软骨肥大带较强,正常对照组基本呈阴性反应.结论MMP-13参与了TMJ创伤所致TMJOA过程,并起了重要作用.

  18. Influence and Significance of Venlafaxine on Basic Fibroblast Growth Factor(bFGF),Vascular Endothelialgrowth Factor (VEGF),Matrix Metalloproteinases-9(MMP-9)Levels in the First-episode Patients with Major Depressive Disorder%文拉法辛对首发抑郁障碍患者血清bFGF、VEGF、MMP-9的影响及意义

    Institute of Scientific and Technical Information of China (English)

    韩毅; 陈涛平; 王丽莉; 左津淮

    2015-01-01

    目的探讨文拉法辛对首发抑郁障碍患者血清碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)、血管内皮细胞生长因子(vascular endothelialgrowth factor,VEGF)、基质金属蛋白酶-9(matrix metal oproteinases-9,MMP)水平的影响及意义。方法采用酶联免疫(ELISA)方法检测38例抑郁患者文拉法辛治疗前及治疗4w后和34名正常对照bFGF、VEGF、MMP-9水平;采用24项汉密尔顿抑郁量表(HAMD24)、汉密尔顿焦虑量表(HMMA)评定治疗前及治疗4w后的抑郁、焦虑症状,应用TESS副反应量表记录药物副反应。结果实验组血清bFGF、VEGF、MMP-9水平治疗前及治疗4w后差异无统计学意义(跃0.05)但均高于对照组,差异有统计学意义(0.05). The cor elation coef icients between the serum bFGF,VEGF,MMP-9 in level and total scores of HAMD-24 and HAMA in patients were not significant ( >0.05). Conclusion bFGF, VEGF, MMP-9 may be involved in the pathophysiology of depression.

  19. 血液灌流对急性百草枯中毒兔氧化应激和基质金属蛋白酶及其抑制物的影响%Effects of hemoperfusion on oxidative stress, matrix metalloproteinases and their inhibitors in acute paraquat poisoning rabbits

    Institute of Scientific and Technical Information of China (English)

    邱俏檬; 刘瑶; 宋芹; 葛赟; 洪广亮; 卢中秋; 梁欢; 李萌芳

    2012-01-01

    the effects of hemoperfusion on oxidative stress status and the levels of matrix metallo proteinase (MMP-2,MMP-9),tissue inhibitor of metalloproteinase (TIMP-1) in lungs,livers and kidneys in paraquat poisoning rabbits,and to explore the mechanism of therapeutic effects induced by HP on acute paraquat poisoning.Methods Seveuty eight rabbits were randomly divided into normal control group (N group,n=6),exposure groups (PQ group,n=24),hemoperfusion treatment group (HP treatment group,n=24) and blank control group (HP group,n=24).The PQ,HPQ and HP groups were divided into 4 observation time groups ( 1,3,7 and 21 d).N group was exposed to 5 ml normal saline and PQ group was exposed to 50 mg/ kg PQ by oral gavage.In 1 h after PQ exposure,HPQ group was exposed to the activated carbon hemoperfusion for 2 h.The content or activity of MDA,SOD and GSH-Px in lungs,livers and kidneys were detected,the expression levels of MMP-2,MMP-9 and TIMP-1 were measured with immunohistochemical SP method for all groups.Results The contents of MDA in lungs,livers and kidneys of PQ and HPQ groups decreased and the activities of SOD and GSH-Px in lungs,livers and kidneys of PQ and HPQ groups increased with observation time.The expression levels of MMP-2,MMP-9 and TIMP-1 in PQ and HPQ groups enhanced on the first day,PQ group was most obvious.Along with the observation time extended,all kinds of positive expression were still high.Compared with normal control group,the activities of serum SOD and GSH-Px in PQ and HPQ groups declined significantly,but the contents of serum MDA increased; the expression levels of MMP-2,MMP-9 and TIMP-1 in lung,liver and kidney tissues increased obviously,the ration between MMP-9 and TIMP-1 significantly increased (P<0.05).Compared with PQ group,the activities of SOD and GSH-Px in HPQ group significantly increased,the content of MDA declined,the expression levels of MMP-2,MMP-9 and TIMP-1 in lung,liver and kidney tissues declined obviously,the ration between MMP

  20. The expressions of MMP-2,MMP-7,MMP-9 and EMMPRIN in human colorectal carcinoma and its significance%结、直肠癌中MMP-2、MMP-7、MMP-9与EMMPRIN 的表达及意义

    Institute of Scientific and Technical Information of China (English)

    凌林; 胡郁之; 孙礼侠; 刘昌阔; 刘志刚

    2015-01-01

    目的::探讨基质金属蛋白酶( MMP)-2、MMP-7、MMP-9和基质金属蛋白酶诱导因子( EMMPRIN)在结、直肠癌中的表达以及在结、直肠癌发展过程中的作用。方法:采用免疫组织化学法检测100例结、直肠癌组织中MMP-2、MMP-7、MMP-9和EMMPRIN的表达。结果:MMP-2、MMP-7、MMP-9和EMMPRIN在结、直肠癌组织中的表达在结、直肠癌的浸润深度、Dukes分期和有无淋巴结转移间差异均有统计学意义(P<0.01);EMMPRIN分别与MMP-2、MMP-7、MMP-9蛋白在结、直肠癌组织中的表达呈正相关关系(P<0.01)。结论:MMP-2、MMP-7、MMP-9和EMMPRIN在结、直肠癌的演进过程中可能具有一定的协同作用。%Objective:To investigate the expressions of matrix metalloproteinases-2,7 and 9(MMP-2,MMP-7 and MMP-9) and extracellular matrix metalloproteinase inducer ( EMMPRIN ) and their roles in the development process of colorectal carcinoma. Methods:The protein expressions of MMP-2,MMP-7,MMP-9 and EMMPRIN in 100 colorectal carcinoma tissue samples were detected by immunohistochemical method. Results:The differences of the expressions rates of MMP-2, MMP-7, MMP-9 and EMMPRIN in colorectal carcinoma tissue samples with different invasion depth,Dukes stage and lymph node metastasis were statistically significant (P<0. 05). The expressions between EMMPRIN and MMP-2,MMP-7 and MMP-9 in colorectal carcinoma tissue were significant positive correlation,respectively(P<0. 01). Conclusions:The expressions of MMP-2,MMP-7,MMP-9 and EMMPRIN have synergetic effects in the development of colorectal carcinoma.

  1. The NAD-Dependent Deacetylase Sirtuin-1 Regulates the Expression of Osteogenic Transcriptional Activator Runt-Related Transcription Factor 2 (Runx2 and Production of Matrix Metalloproteinase (MMP-13 in Chondrocytes in Osteoarthritis

    Directory of Open Access Journals (Sweden)

    Koh Terauchi

    2016-06-01

    Full Text Available Aging is one of the major pathologic factors associated with osteoarthritis (OA. Recently, numerous reports have demonstrated the impact of sirtuin-1 (Sirt1, which is the NAD-dependent deacetylase, on human aging. It has been demonstrated that Sirt1 induces osteogenic and chondrogenic differentiation of mesenchymal stem cells. However, the role of Sirt1 in the OA chondrocytes still remains unknown. We postulated that Sirt1 regulates a hypertrophic chondrocyte lineage and degeneration of articular cartilage through the activation of osteogenic transcriptional activator Runx2 and matrix metalloproteinase (MMP-13 in OA chondrocytes. To verify whether sirtuin-1 (Sirt1 regulates chondrocyte activity in OA, we studied expressions of Sirt1, Runx2 and production of MMP-13, and their associations in human OA chondrocytes. The expression of Sirt1 was ubiquitously observed in osteoarthritic chondrocytes; in contrast, Runx2 expressed in the osteophyte region in patients with OA and OA model mice. OA relating catabolic factor IL-1βincreased the expression of Runx2 in OA chondrocytes. OA chondrocytes, which were pretreated with Sirt1 inhibitor, inhibited the IL-1β-induced expression of Runx2 compared to the control. Since the Runx2 is a promotor of MMP-13 expression, Sirt1 inactivation may inhibit the Runx2 expression and the resultant down-regulation of MMP-13 production in chondrocytes. Our findings suggest thatSirt1 may regulate the expression of Runx2, which is the osteogenic transcription factor, and the production of MMP-13 from chondrocytes in OA. Since Sirt1 activity is known to be affected by several stresses, including inflammation and oxidative stress, as well as aging, SIRT may be involved in the development of OA.

  2. Matrix metalloproteinase/tissue inhibitor of metalloproteinase,vascular endothelial growth factor with invasion and metastasis of lung cancer%MMP/TIMP、VEGF与肺癌的浸润和转移

    Institute of Scientific and Technical Information of China (English)

    赵志芳; 陈刚

    2007-01-01

    肺癌的浸润和转移与基质金属蛋白酶(matrix metalloproteinase,MMP)及其抑制剂基质金属蛋白酶组织抑制剂(tissue inhibitor of metalloproteinase,TIMP)、血管内皮生长因子(vascular endothelial growth factor,VEGF)等多因素密切相关,深入研究肺癌中MMP/TIMP及VEGF的表达与功能,揭示肺癌的浸润与转移的机制,可为评估肺癌的浸润、转移及预后提供重要的参考指标.

  3. A novel marker for assessment of liver matrix remodeling: An enzyme-linked immunosorbent assay (ELISA) detecting a MMP generated type I collagen neo-epitope (C1M)

    DEFF Research Database (Denmark)

    Leeming, Diana Julie; He, Y.; Veidal, S. S.

    2011-01-01

    A competitive enzyme-linked immunosorbent assay (ELISA) for detection of a type I collagen fragment generated by matrix metalloproteinases (MMP) -2, -9 and -13, was developed (CO1-764 or C1M). The biomarker was evaluated in two preclinical rat models of liver fibrosis: bile duct ligation (BDL) an......-764 marker was not correlated with skeletal involvement or number of bone metastases. This ELISA has the potential to assess the degree of liver fibrosis in a non-invasive manner.......) and carbon tetra chloride (CCL4)-treated rats. The assay was further evaluated in a clinical study of prostate-, lung-and breast-cancer patients stratified according to skeletal metastases. A technically robust ELISA assay specific for a MMP-2, -9 and -13 neo-epitope was produced and seen to be statistically...

  4. MMP2 — EDRN Public Portal

    Science.gov (United States)

    MMP2 is a member of the matrix metalloproteinase (MMP) family and is involved in many functions, such as remodeling of the vasculature, angiogenesis, tissue repair and remodeling, tumor invasion, inflammation, atherosclerotic plaque rupture, reproduction and embryonic development, as well as in disease processes such as arthritis and metastasis. In addition to degrading extracellular matrix proteins, MMP2 can also act on several nonmatrix proteins such as big endothelial 1 and beta-type CGRP promoting vasoconstriction and appears to have a role in myocardial cell death pathways. MMPs are generally secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The MMP2 protein degrades gelatin type I and collagen types IV, V, VII, and X. MMP2 gene mutations have been associated with Winchester syndrome and Nodulosis-Arthropathy-Osteolysis (NAO) syndrome. There are two known isoforms of this gene, encoded by two transcript variants.

  5. The Oncogenic Response to MiR-335 Is Associated with Cell Surface Expression of Membrane-Type 1 Matrix Metalloproteinase (MT1-MMP) Activity.

    Science.gov (United States)

    Rojas, Fausto; Hernandez, Maria E; Silva, Milagros; Li, Lihua; Subramanian, Subbaya; Wilson, Michael J; Liu, Ping

    2015-01-01

    MicroRNA miR-335 has been reported to have both tumor suppressor and oncogenic activities. In order to determine possible tissue and cell type differences in response to miR-335, we examined the effect of miR-335 on cell expression of MT1-MMP, a proteinase commonly expressed in tumors and associated with cell proliferation and migration. miR-335 increased cell surface expression of MT1-MMP in fibrosarcoma HT-1080 and benign prostate BPH-1 cells, but not in prostate LNCaP or breast MCF-7 tumor cells. miR-335 stimulated proliferation and cell migration in a wound healing in vitro assay in HT-1080, BPH-1, and U87 glioblastoma cells, cells which demonstrated significant cell surface expression of MT1-MMP. In contrast, miR-335 did not affect proliferation or migration in cells without a prominent plasma membrane associated MT1-MMP activity. Our data suggest that differences in response to miR-335 by tumor cells may lie in part in the mechanism of regulation of MT1-MMP production.

  6. The Oncogenic Response to MiR-335 Is Associated with Cell Surface Expression of Membrane-Type 1 Matrix Metalloproteinase (MT1-MMP Activity.

    Directory of Open Access Journals (Sweden)

    Fausto Rojas

    Full Text Available MicroRNA miR-335 has been reported to have both tumor suppressor and oncogenic activities. In order to determine possible tissue and cell type differences in response to miR-335, we examined the effect of miR-335 on cell expression of MT1-MMP, a proteinase commonly expressed in tumors and associated with cell proliferation and migration. miR-335 increased cell surface expression of MT1-MMP in fibrosarcoma HT-1080 and benign prostate BPH-1 cells, but not in prostate LNCaP or breast MCF-7 tumor cells. miR-335 stimulated proliferation and cell migration in a wound healing in vitro assay in HT-1080, BPH-1, and U87 glioblastoma cells, cells which demonstrated significant cell surface expression of MT1-MMP. In contrast, miR-335 did not affect proliferation or migration in cells without a prominent plasma membrane associated MT1-MMP activity. Our data suggest that differences in response to miR-335 by tumor cells may lie in part in the mechanism of regulation of MT1-MMP production.

  7. The clinical significance of Matrix Metalloproteinase-13 and Galectin-3 in synovial tissues of osteoarthritis w%MMP-13和 Galectin-3在骨性关节炎滑膜组织中的临床意义

    Institute of Scientific and Technical Information of China (English)

    马丽艳; 宋旭东; 张德新; 程潭

    2013-01-01

      目的检测 MMP-13和 Galectin-3在骨性关节炎滑膜组织中的表达,探讨二者在骨性关节炎发病机制中的作用。方法通过关节镜获取42例骨性关节炎滑膜标本,同时选择12例非正常死亡及外伤性截肢的健康青年的滑膜组织作为对照组。采用免疫组化检测 MMP-13及 Galectin-3在骨性关节炎滑膜组织中的表达。结果病理观察显示骨性关节炎滑膜细胞增生,淋巴细胞及浆细胞浸润,甚至形成淋巴滤泡,同时微血管增生,管壁增厚。免疫组化结果显示:在骨性关节炎滑膜组织中,MMP-13蛋白主要表达于滑膜衬里层细胞、炎细胞、巨噬细胞样组织细胞以及少许血管内皮细胞。在正常滑膜组织中,MMP-13蛋白则主要表达于滑膜衬里层细胞。二者的阳性细胞率分别为0.387±0.042和0.053±0.019,前者的阳性表达率明显高于后者( P<0.05)。Galectin-3蛋白在骨性关节炎组中的阳性表达主要分布于滑膜衬里层细胞及滑膜下层少许炎细胞中。Galectin-3蛋白在骨性关节炎组和对照组的阳性细胞率分别为0.267±0.044和0.046±0.021。前者的阳性表达率明显高于后者( P<0.05)。MMP-13和 Galectin-3在骨性关节炎滑膜中表达与患者年龄及性别无关( P>0.05),而与骨性关节炎X线片临床分期有关,MMP-3与 Galectin-3在滑膜中的表达均随着骨性关节炎 X线片临床分期的增高而增强( P<0.05)。结论MMP-13蛋白和 Galectin-3蛋白在骨性关节炎滑膜组织中的表达显著高于正常滑膜组织,随X线片临床分期的增高而增强,表明二者参与骨性关节炎滑膜的病理过程,对滑膜炎起相互促进作用,临床检测滑膜组织中 MMP-13蛋白和 Galectin-3蛋白的表达有助于指导骨性关节炎的治疗及病情的评估。%Objective To detect the protein expressions of Matrix Metalloproteinase-13 ( MMP-13 ) and Galectin-3 in synovial tissues of

  8. MMP10 — EDRN Public Portal

    Science.gov (United States)

    From NCBI Gene: Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The enzyme encoded by this gene degrades proteoglycans and fibronectin. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3. [provided by RefSeq, Jul 2008

  9. Functional Promoter Polymorphisms of MMP-2 C-735T and MMP-9 C-1562T and Their Synergism with MMP-7 A-181G in Multiple Sclerosis.

    Science.gov (United States)

    Rahimi, Zohreh; Abdan, Zahra; Rahimi, Ziba; Razazian, Nazanin; Shiri, Hadis; Vaisi-Raygani, Asad; Shakiba, Ebrahim; Vessal, Mahmood; Moradi, Mohammad-Taher

    2016-08-01

    Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system. Matrix metalloproteinases (MMPs) play an important role in breakdown of blood-brain barrier, transmigration, and invasion of immune cells and formation of MS lesions. The aim of present study was to investigate the influence of MMP-2 C-735T and MMP-9 C-1562T variants and their synergism with MMP-7 A-181G on susceptibility to MS. In a case-control study 125 MS patients and 235 healthy individuals from Western Iran were investigated. The various genotypes of MMP-2, MMP-9, and MMP-7 were detected using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In females the presence of MMP-2 C allele was associated with an increased risk of MS (OR = 1.69, p = 0.041). No significant difference was detected between the frequency of MMP-9 T allele in MS patients (8.2%) and controls (12.8%, p = 0.068). The concomitant presence of both MMP-2 C and MMP-7 G alleles was associated with 1.82-fold increased risk of MS (p = 0.002). Also, a synergism was detected between MMP-9 C and MMP-7 G alleles that elevated the risk of MS by 1.5-times (p = 0.035). The presence of haplotype MMP-9 T, MMP-7 G, and MMP-2 C (TGC) compared to haplotype CAG increased the risk of MS by 3.13-fold (p = 0.16). The present study suggests that gene-gene interactions and variants of more genes instead of single gene might play a role in susceptibility to MS. We indicated that synergism between variants of MMP-2, MMP-7, and MMP-9 genes might increase the risk of MS.

  10. Increased MMP-2 activity during intervertebral disc degeneration is correlated to MMP-14 levels

    NARCIS (Netherlands)

    Rutges, J. P. H. J.; Kummer, J. A.; Oner, F. C.; Verbout, A. J.; Roestenburg, H. J. A.; Dhert, W. J. A.; Creemers, L. B.

    2008-01-01

    Intervertebral disc (IVD) degeneration is associated with the increased expression of several matrix metalloproteinases (MMPs), in particular MMP-2. However, little is known about the actual activity of MMP-2 in healthy and degenerated discs, or what mechanisms are involved in its activation. A majo

  11. Molecular Modeling and Docking Studies on the First Chlorotoxin-Like Peptide from Iranian Scorpion Mesobuthuseupeus (Meict and SNP Variants of Matrix Methaloproteinase-2 (MMP-2

    Directory of Open Access Journals (Sweden)

    Farzaneh Mohammadi Farsani

    2015-09-01

    Full Text Available Background: MeICT is the first chlorotoxin-like peptide isolated from the Iranian Scorpion Mesobuthus eupeus. Chlorotoxin (CTX is a neurotoxin that specially binds to (MMP-2 on ma-lignant cells and now is used in treatment of glioma. In the present study, we have used homology modeling to propose the 3D structure of MeICTand analyze its interaction with MMP-2 and its SNP types. Methods: The structure of MeICT was modeled by using homology modeling through the Swiss-Model workspace. Structural evaluation and stereo-chemical analysis of modeled struc-ture of MeICT was performed using ProSA-web Z-scores and Mol Probity Ramachandran plots. Hex Server was used to investigate the interactions between MeICT and catalytic domain of MMP-2 and SNP types. Binding energies calculation and complementarity scores were used for evaluation of protein docking. Results:The comparable Z-scores, Ramachandran plot characteristics and RMSD values confirmed the quality of the homology model of MeICT. About 17 SNP variants in catalytic domain of MMP2 were detected. According to the total and electrostatic energies and the number of interactive residues by hydrogen bond, the structure of MeICT-rs200271857, MeICT-rs144334568, MeICT-rs111590299 and MeICT-rs201083413complexes are more stable. Conclusion: The structure of MeICT is similar to CTX, somight be used as therapeutic agent in glioma. We could find some variants of MMP-2 that can bind to MeICT with more or less af-finity and can affect treatment pathway.

  12. Genetic Association of MMP10, MMP14, and MMP16 with Dental Caries

    Science.gov (United States)

    Feingold, E.; Cooper, M.; Vanyukov, M. M.; Maher, B. S.; Slayton, R. L.; Willing, M. C.; Reis, S. E.; Crout, R. J.; Weyant, R. J.; Levy, S. M.; Marazita, M. L.

    2017-01-01

    Matrix metalloproteinases (MMPs), which degrade extracellular proteins as part of a variety of physiological processes, and their inhibitors have been implicated in the dental caries process. Here we investigated 28 genetic variants spanning the MMP10, MMP14, and MMP16 genes to detect association with dental caries experience in 13 age- and race-stratified (n = 3,587) samples from 6 parent studies. Analyses were performed separately for each sample, and results were combined across samples by meta-analysis. Two SNPs (rs2046315 and rs10429371) upstream of MMP16 were significantly associated with caries in an individual sample of white adults and via meta-analysis across 8 adult samples after gene-wise adjustment for multiple comparisons. Noteworthy is SNP rs2046315 (p = 8.14 × 10−8) association with caries in white adults. This SNP was originally nominated in a genome-wide-association study (GWAS) of dental caries in a sample of white adults and yielded associations in a subsequent GWAS of surface level caries in white adults as well. Therefore, in our study, we were able to recapture the association between rs2046315 and dental caries in white adults. Although we did not strengthen evidence that MMPs 10, 14, and 16 influence caries risk, MMP16 is still a likely candidate gene to pursue. PMID:28348596

  13. The relationship between matrix metalloproteinases-9 and CD15s antigen in angiogenesis of gastric carcinoma%MMP-9和CD15s与胃癌血管生成的相关性研究及其临床意义

    Institute of Scientific and Technical Information of China (English)

    Peizhong Shang; Xiaowu Li; Xiqin Zhu; Jianjun Miao; Guohong Jia; Huaping Gu

    2009-01-01

    Objective: To investigate the relationship between matrix metalloproteinases-9 (MMP-9) and sialyl Lewis X (CD15s) antigen in invasion and metastasis of gastric carcinoma. Methods: Expression of CD105, MMP-9 and CD15s in 47 cases of gastric carcinoma undergone radical surgery were evaluated by SP immunohistochemical staining using the respective monoclonal antibody. The microvessel density (MVD) marked with CD105 was detected. Correlation between MVD and MMP-9, CD15s was also statistically analyzed. Results: The MVD in both MMP-9 and CD15s positive expression group was higher significantly than that in MMP-9 or CD15s positive expression alone group, and it was also significantly higher than that in both MMP-9 and CD15s negative expression group. Conclusion: MMP-9 is a marker of invasion and CD15s is a marker of metastasis in gastric carcinoma. Combining detection of MMP-9 and CD15s has certain clinical significance for diagnosis,treatment and assessing the prognosis for gastric caner.

  14. MMP-12、Timp-1、Elastin在女性压力性尿失禁患者阴道壁组织中的表达%Expressions of matrix metalloproteinase-12,tissue inhibitor of matrix metallopro-teinase-1 and Elastin in vaginal wall tissues of women with stress urinary incontinence

    Institute of Scientific and Technical Information of China (English)

    张晓丹; 李晓果; 许雅娟; 张彩霞; 王鲁文

    2011-01-01

    Objective: To explore the expressions of matrix metalloproteinase - 12 ( MMP - 12), tissue inhibitor of matrix metalloproteinase- 1 (Timp- 1 ) and Elastin in vaginal wall tissues of women with stress urinary incontinence (SUI), and their relationship with occurrence of SUI. Methods: 30 samples of vaginal wall connective tissues of women with SUI were selected as study group, and 30 samples of vaginal wall connective tissues of women with pelvic organ prolapse (POP) were selected as POP group, 30 women without SUI and POP were selected as control group, immunohistochemical staining was used in the study, the expression levels of MMP - 12, Timp - 1 and Elastin were detected, PCR was used to detect the expression of Elastin mRNA in vaginal wall tissues of the three groups. Results: The expression level of MMP - 12 in SUI group was significantly higher than that in control group (P < 0. 05 ), the expression levels of Timp - 1 and Elastin in SUI group was significantly lower than those in control group ( P < 0. 05 ), but there was no significant difference in expression levels of MMP - 12, Timp - 1 and Elastin between SUI group and POP group, the expression results of Elastin detected by immanohistochemical staining and PCR were coincident. Conclusion: The expression level of MMP - 12 is high in vaginal wall tissues of women with SUI, Timp -1 is an inhibitor of MMP - 12; the expression level of Timp - 1 is low, which increases the degradation of Elastin, reduces the Elastin fibers,then the elasticity of pelvic floor tissues decreases.%目的:探讨基质金属蛋白酶(MMP-12)、基质金属蛋白酶抑制剂(Timp-1)、弹性蛋白(Elastin)在女性压力性尿失擎(SUI)患者阴道壁巾的表达及三者与SUI发病的关系.方法:研究组(SUI组)为30例有完整随访资料的女性SUI患者阴迫壁结缔组织标本,盆底器官脱垂组(POP组)为30例有完整随访资料的女性盆底器官脱垂(POP)患者的阴道壁结缔组织标本,对照组为30

  15. Expression of Matrix Metalloproteinase in Cervical Squamous Cell Carcinoma Tissue and Its Significance%子宫颈鳞癌组织中基质金属蛋白酶MMP-2和MMP-9的表达及其意义

    Institute of Scientific and Technical Information of China (English)

    张淑兰; 林蓓; 蔡威; 牛菊敏

    2003-01-01

    目的:探讨基质金属蛋白酶MMP-2和MMP-9与宫颈鳞癌的发生、发展及转移的关系.方法:免疫组化S-P法测定MMP-2和MMP-9表达及分布;明胶酶谱法测定活性型MMP-2及MMP-9蛋白的含量;RT-PCR技术检测MMP-2及MMP-9 mRNA表达水平.结果:宫颈癌组织中MMP-2及MMP-9的阳性表达率为77.78%和66.67%,明显高于在CIN(10%及20%)及正常宫颈组织中表达率.淋巴转移组MMP-2及MMP-9的阳性率均显著高于非淋巴转移组,P<0.05.在宫颈鳞癌组织中MMP-2及MMP-9酶活性显著高于CIN及正常宫颈组织,P<0.01.MMP-2及MMP-9 mRNA在宫颈鳞癌中的表达量亦显著高于在CIN及正常宫颈组织的表达量,P<0.01.结论:MMP-2及MMP-9与宫颈鳞癌的发生、发展及转移有关,MMP-2及MMP-9的增高可作为判断宫颈鳞癌具有转移倾向的临床参考指标.

  16. MMP-1 polymorphism and its relationship to pathological processes

    Indian Academy of Sciences (India)

    P A Arakaki; M R Marques; M C L G Santos

    2009-06-01

    Matrix metalloproteinases (MMPs) are a family of zinc (Zn)-dependent endopeptidases that are collectively capable of cleaving virtually all extracellular matrix (ECM) substrates and play an important role in diverse physiological and pathological processes. The activity of MMPs is regulated at multiple levels. The transcriptional regulation of MMP appears to represent the key step in MMP regulation. There are diverse types of MMPs that differ structural and functionally. MMP-1 is the most ubiquitously expressed interstitial collagenase and has a prominent role in initial cleavage of the ECM. The level of MMP-1 expression can be influenced by different single-nucleotide polymorphisms (SNPs) in the promoter region. A functional polymorphism at position –1607 has been shown to alter the transcriptional activity of MMP-1 and was associated with diverse pathological processes. The aim of our review was to discuss some topics related to MMP in physiological and pathological processes, with a focus on MMP-1 polymorphism.

  17. Genistein Suppression of Matrix Metalloproteinase 2 (MMP-2) and Vascular Endothelial Growth Factor (VEGF) Expression in Mesenchymal Stem Cell Like Cells Isolated from High and Low Grade Gliomas

    Science.gov (United States)

    Yazdani, Yasaman; Sharifi Rad, Mohammad Reza; Taghipour, Mousa; Chenari, Nooshafarin; Ghaderi, Abbas; Razmkhah, Mahboobeh

    2016-12-01

    Objective: Brain tumors cause great mortality and morbidity worldwide, and success rates with surgical treatment remain very low. Several recent studies have focused on introduction of novel effective medical therapeutic approaches. Genistein is a member of the isoflavonoid family which has proved to exert anticancer effects. Here we assessed the effects of genistein on the expression of MMP-2 and VEGF in low and high grade gliomas in vitro. Materials and Methods: High and low grade glioma tumor tissue samples were obtained from a total of 16 patients, washed with PBS, cut into small pieces, digested with collagenase type I and cultured in DMEM containing 10% FBS. When cells reached passage 3, they were exposed to genistein and MMP-2 and VEGF gene transcripts were determined by quantitative real time PCR (qRT-PCR). Results: Expression of MMP-2 demonstrated 580-fold reduction in expression in low grade glioma cells post treatment with genistein compared to untreated cells (P value= 0.05). In cells derived from high grade lesions, expression of MMP-2 was 2-fold lower than in controls (P value> 0.05). Genistein caused a 4.7-fold reduction in VEGF transcript in high grade glioma cells (P value> 0.05) but no effects were evident in low grade glioma cells. Conclusion. Based on the data of the present study, low grade glioma cells appear much more sensitive to genistein and this isoflavone might offer an appropriate therapeutic intervention in these patients. Further investigation of this possibility is clearly warranted.

  18. MMP7 — EDRN Public Portal

    Science.gov (United States)

    Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, and tissue remodeling, as well as in disease processes, such as arthritis and metastasis. Most MMP's are secreted as inactive proproteins which are activated when cleaved by extracellular proteinases. The enzyme encoded by this gene degrades proteoglycans, fibronectin, elastin and casein and differs from most MMP family members in that it lacks a conserved C-terminal protein domain. The enzyme is involved in wound healing, and studies in mice suggest that it regulates the activity of defensins in intestinal mucosa. The gene is part of a cluster of MMP genes which localize to chromosome 11q22.3.

  19. Imagerie moleculaire de la MMP-2

    Science.gov (United States)

    Lebel, Rejean

    MMPs (matrix metalloproteinases) are enzymes involved in tissue architecture remodelling and cell migration. MMP-2, particularly, was found to be a biomarker of the progression or prognosis of several pathologies, such as arthritis, atherosclerosis, infarct and cancer. Yet, its exact role in these pathologies is still uncertain. For these reasons, it is critical to develop new tools to enable the specific and non invasive study of MMP-2. As of now, a large number of optical probes (optical imaging), contrast agents (magnetic resonance imaging) and radiotracers (positron emission tomography, single photon emission tomography) have been published in the literature. However, none of the molecules allows for the specific quantification of MMP-2, particularly against MMP-9 which cleaves similar substrates. This thesis describes our progress in the development of new molecules capable of targeting and allowing the imaging of MMP-2. All tested molecules were fond to be quickly activated by MMP-2 and to be selective when compared with MMP3, MMP-7 and MMP-9. First, a contrast agent named PCA2-switch is tested in a mouse subcutaneous tumor model, and allows us to differentiate between tumors with low or high levels of MMP-2 activity. We also developed a panel of activatable fluorescent probes, one of which was found to be highly specific to MMP-2 (low activation by MMP9, efficient quenching). However, an extensive set of control experiments does not enable to conclude on the specificity of the probes in vivo. One of the principal limitations of many studies in the field of MMP imaging is the lack of proper controls, including unspecific uptake controls. The last section of this thesis discusses the impact of the EPR (enhanced permeability and retention) effect on the uptake of a non-specific probe in a subcutaneous tumor model treated with radiotherapy. Proper control experiments that should be performed when testing new MMP-2 probes are discussed. Keywords: Matrix

  20. Association of MMP-8 with obesity, smoking and insulin resistance.

    Science.gov (United States)

    Lauhio, Anneli; Färkkilä, Esa; Pietiläinen, Kirsi H; Åström, Pirjo; Winkelmann, Alina; Tervahartiala, Taina; Pirilä, Emma; Rissanen, Aila; Kaprio, Jaakko; Sorsa, Timo A; Salo, Tuula

    2016-09-01

    Obesity has been recognized as a state of subclinical inflammation resulting in a loss of insulin receptors and decreased insulin sensitivity. We here studied in vivo the role of circulating matrix metalloproteinase-8 (MMP-8) among young healthy twin adults. Also, in vitro analysis of the cleavage of human insulin receptor (INSR) by MMP-8 was investigated as well its inhibition by doxycycline and other MMP-8 inhibitor, Ilomastat/GM6001, which are broad-spectrum MMP inhibitors. We analysed serum MMP-8 levels by a time-resolved immunofluorometric assay in obese (n = 34), overweight (n = 76) and normal weight (n = 130) twin individuals. The effect of MMP-8 on INSR and the effects of synthetic MMP-8 inhibitors, doxycycline and Ilomastat/GM6001, were studied by SDS-PAGE. We found that in obese individuals relative to normal weight individuals, the serum MMP-8 levels and MMP-8/TIMP-1 ratio were significantly increased (P = 0·0031 and P = 0·031, respectively). Among normal weight and obese individuals, also smoking significantly increases serum MMP-8 and MMP-8/TIMP-1 ratio. In vitro, we found that INSR was degraded by MMP-8 and this was inhibited by doxycycline and Ilomastat/GM6001. Obesity associated with elevated circulating MMP-8 found among young adults may contribute to progression of insulin resistance by cleaving INSR. This INSR cleavage by MMP-8 can be inhibited by synthetic MMP-8 inhibitors such as doxycycline. In addition to obesity, also smoking independently explained increased MMP-8 levels. Our results suggest that MMP-8 is an essential mediator in systemic subclinical inflammatory response in obesity, and a potential drug target. © 2016 Stichting European Society for Clinical Investigation Journal Foundation.

  1. MMP-2、MMP-9及其抑制因子TIMP-1、TIMP-2在宫颈癌中的表达%Expression and significance of matrix metalloproteinase and tissu inhibitor of matrix metalloproteinase in cervical cancer

    Institute of Scientific and Technical Information of China (English)

    游泳; 杜莹莹; 曹媛; 李真珍; 张胜军

    2014-01-01

    目的 分析基质金属蛋白酶2(MMP-2)和MMP-9及其抑制因子1(TIMP-1)和TIMP-2在宫颈癌不同位置中的表达情况及其临床意义.方法 选取经病理证实的宫颈浸润癌患者118例(ICC组)、宫颈上皮内瘤样病变患者75例(CIN组),取病变中心组织和边缘组织;选取正常宫颈组织标本45例为对照组.采用SABC法行免疫组化检测各组中MMP-2、MMP-9、TIMP-1和TIMP-2因子表达阳性率、染色强度和表达强度.比较各组相关因子表达情况差异.结果 ICC组中MMP-2和MMP-9的阳性率、染色强度和表达强度显著高于CIN组和对照组,TIMP-1和TIMP-2的阳性率、染色强度和表达强度显著低于CIN组和对照组(P<0.05).在ICC组,边缘癌组织的MMP-2与MMP-9的阳性率、染色强度和表达强度明显高于中心癌组织,而TIMP-1与TIMP-2的阳性率、染色强度和表达强度明显低于中心癌组织(P<0.05).结论 MMP-2、MMP-9及其抑制因子TIMP-1、TIMP-2与宫颈癌的发生、发展密切相关,可能在宫颈癌的侵袭与转移中发挥着重要作用.

  2. Inhibitory effect of the carnosine-gallic acid synthetic peptide on MMP-2 and MMP-9 in human fibrosarcoma HT1080 cells.

    Science.gov (United States)

    Kim, Sung-Rae; Eom, Tae-Kil; Byun, Hee-Guk

    2014-09-01

    Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases that degrade extracellular matrix components and play important roles in a variety of biological and pathological processes such as malignant tumor metastasis and invasion. In this study, we constructed carnosine-gallic acid peptide (CGP) to identify a better MMP inhibitor than carnosine. The inhibitory effects of CGP on MMP-2 and MMP-9 were investigated in the human fibrosarcoma (HT1080) cell line. As a result, CGP significantly decreased MMP-2 and MMP-9 expression levels without a cytotoxic effect. Moreover, CGP may inhibit migration and invasion in HT1080 cells through the urokinase plasminogen activator (uPA)-uPA receptor signaling pathways to inhibit MMP-2 and MMP-9. Based on these results, it appears that CGP may play an important role in preventing and treating several MMP-2 and MMP-9-mediated health problems such as metastasis.

  3. Detection and the clinical significance of serum MMP-2、MMP-3、MMP-9 and TIMP-4 levels in patients with systemic lupus erythematosus%系统性红斑狼疮患者外周血MMP-2、MMP-3、MMP-9和TIMP-4水平的检测及其临床意义

    Institute of Scientific and Technical Information of China (English)

    胡亮; 彭奕冰; 王学锋; 巩惠芸

    2012-01-01

    目的:检测系统性红斑狼疮(SLE)患者外周血基质金属蛋白酶(MMP)-2、MMP-3、MMP-9和基质金属蛋白酶组织型抑制因子(TIMP)-4的水平,并探讨其临床意义.方法:采用双抗夹心ELISA法,检测58例SLE患者及30例正常对照者的血清MMP-2、MMP-3、MMP-9和TIMP-4水平.结果:SLE患者血清MMP-3、TIMP-4水平显著高于正常对照者,但其血清MMP-2、MMP-9水平则与正常对照者间无统计学差异.SLE患者的血清MMP-3水平与MMP-2、MMP-9、TIMP-4水平均呈正相关,MMP-9水平与MMP-2水平间亦呈正相关;同时其MMP-2、MMP-3、MMP-9水平与反映肾脏、肝脏及机体免疫状态的多组指标间存在相关性,包括尿素氮、肌酐、尿酸、白蛋白、免疫球蛋白G、免疫球蛋白A、免疫球蛋白M、补体C3和补体C4等;SLE患者中,发生血小板减少者的血清MMP-2、MMP-9水平与血小板正常者相比显著降低.结论:SLE患者外周血MMP-3和TIMP-4水平显著升高.不同MMP间可相互调节,同时也受TIMP-4等TIMP的调节.MMP-2、MMP-3和MMP-9可能参与了SLE患者肾脏和肝脏的病理损害过程.%Objective To detect serum matrix metalloproteinase (MMP)-2, MMP-3, MMP-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-4 levels in patients with systemic lupus erythematosus (SLE) and define its clinical significance. Methods Serum levels of MMP-2, MMP-3, MMP-9 and TIMP-4 were measured by ELISA. Results Levels of MMP-3 and TIMP-4 were significantly higher in SLE patients than those in healthy controls. Positive correlation was found between these MMPs. Serum MMP-2, MMP-3, MMP-9 levels had a correlation with other laboratory results reflecting the condition of kidney, liver and immunity. MMP-2 and MMP-9 levels were decreased in SLE patients having a low platelet count. Conclusins Serum MMP-2 and TIMP-4 levels are significantly higher in patients with SLE. MMP-2, MMP-3 and MMP-9 were regulated by a complicated network. TIMP-4 is involved in this

  4. Role of MMP-3 and MMP-9 and their haplotypes in risk of bladder cancer in North Indian cohort.

    Science.gov (United States)

    Srivastava, Priyanka; Mandhani, Anil; Kapoor, Rakesh; Mittal, Rama D

    2010-11-01

    Matrix metalloproteinases (MMPs) play critical roles in cancer development and progression. Nonsynonymous single nucleotide polymorphisms (SNPs) in functional domain of MMP-3 and MMP-9 contribute appreciably to cancer predisposition and aggression. To test this proposition we examined whether six SNPs of the MMP-3 and MMP-9 genes are associated with risk of bladder cancer (BC) in a North Indian population. Six SNPs of MMP-3 and MMP-9 were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in a case-control study including 200 BC patients and 200 age/gender/ethnicity-matched controls. Increased risk for BC susceptibility was observed in MMP-3 (1171) 5A/5A [P = 0.022; odds ratio (OR), 3.46; 95% confidence interval (CI), 1.20-9.98], MMP-9 (Q279R) QQ (P = 0.048; OR, 1.92; 95%CI, 1.01-3.66), MMP-9 (P574R) PR (P BCG)-treated non-muscle-invasive BC (NMIBC) patients (log-rank P = 0.025). Our data suggested that MMP-3-1171 5A/5A and MMP-9 (Q279R) QQ, MMP-9 (P574R) PR, PR + RR, and R allele are associated with high risk of BC.

  5. Zymographic patterns of MMP-2 and MMP-9 in the CSF and cerebellum of dogs with subacute distemper leukoencephalitis.

    Science.gov (United States)

    Machado, Gisele F; Melo, Guilherme D; Souza, Milena S; Machado, Andressa A; Migliolo, Daniela S; Moraes, Olívia C; Nunes, Cáris M; Ribeiro, Erica S

    2013-07-15

    Distemper leukoencephalitis is a disease caused by the canine distemper virus (CDV) infection. It is a demyelinating disease affecting mainly the white matter of the cerebellum and areas adjacent to the fourth ventricle; the enzymes of the matrix metalloproteinases (MMPs) group, especially MMP-2 and MMP-9 have a key role in the myelin basic protein fragmentation and in demyelination, as well as in leukocyte traffic into the nervous milieu. To evaluate the involvement of MMPs during subacute distemper leukoencephalitis, we measured the levels of MMP-2 and MMP-9 by zymography in the cerebrospinal fluid (CSF) and in the cerebellum of 14 dogs naturally infected with CDV and 10 uninfected dogs. The infected dogs presented high levels of pro-MMP-2 in the CSF and elevated levels of pro-MMP-2 and pro-MMP-9 in the cerebellar tissue. Active MMP-2 was detected in the CSF of some infected dogs. As active MMP-2 and MMP-9 are required for cellular migration across the blood-brain barrier and any interference between MMPs and their inhibitors may result in an amplification of demyelination, this study gives additional support to the involvement of MMPs during subacute distemper leukoencephalitis and suggests that MMP-2 and MMP-9 may take part in the brain inflammatory changes of this disease.

  6. HPA和MMP9对非小细胞肺癌患者临床预后评价的意义研究%Significance of serum heparanase and matrix metalloproteinases 9 in the prognosis of non-small cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    夏彦东; 常延河; 杨宏秀; 刘丽

    2015-01-01

    目的:检测非小细胞肺癌( NSCLC)患者血清HPA和MMP9水平,分析其与NSCLC患者淋巴结转移以及预后的关系。方法选取80例NSCLC患者和30例健康对照,采用酶联免疫法检测HPA和MMP9的血清水平,分析HPA和MMP9与患者临床病理特征的关系,通过受试者工作曲线分析血清HPA和MMP9用于判断NSCLC患者是否发生淋巴结转移的可行性,通过Kaplan-Meier法进行分析HPA和MMP9预测病人预后的临床意义。结果 NSCLC患者血清HPA和MMP9值均高于健康对照组;血清HPA与淋巴结转移、远处转移有关,血清MMP9值与TNM分期、淋巴结转移、远处转移有关;血清HPA和MMP9用于预测NSCLC患者淋巴结转移情况的曲线下面积分别为0.732和0.785;NSCLC患者血清HPA和MMP9越高,患者预后越差。结论血清HPA和MMP9均可用于确定NSCLC患者是否发生淋巴结转移,并可用于评估病人预后。%Objective To detect the level of serum heparanase ( HPA ) and matrix metalloproteinases 9 (MMP9) in non-small cell lung cancer (NSCLC) patients, and to investigate the relationship of the two indexes with lymph node metastasis and prognosis. Methods The serum levels of HPA and MMP9 were detected in 80 NSCLC patients and 30 health controls by enzyme-linked immunosorbent assay ( ELISA) . The clinical and pathologic features were analyzed through the levels of HPA and MMP9. The specificity and sensitivity of serum HPA and MMP9 to de-termine lymph node metastasis or not were analyzed by receiver operating characteristic ( ROC) curve. Kaplan-Meier method was used to evaluate overall survival according to cutoff serum levels. Results The serum levels of HPA and MMP9 were higher in NSCLC patients than in health controls. The serum level of HPA was associations with lymph node metastasis and distance metastasis. The serum level of MMP9 was associated with TNM stage, lymph node me-tastasis and distance metastasis. The area under the curve

  7. MMP-9 and MMP-2 activities in stomach and breast tumours, as measured by a novel MMP activity assay using modified urokinase as a substrate

    NARCIS (Netherlands)

    Hanemaaijer, R.; Visser, H.; Duffy, J.; Verspaget, H.W.; Verheijen, J.H.; Maguire, T.

    1998-01-01

    Matrix metalloproteinases (MMPs) play an important role in many pathological processes. However, MMP activities are difficult to determine since no simple specific and/or chromogenic substrates exist. Therefore, we have developed a novel MMP activity assay using a modified urokinase as a substrate.

  8. Severity of Plasma Leakage Is Associated With High Levels of Interferon γ-Inducible Protein 10, Hepatocyte Growth Factor, Matrix Metalloproteinase 2 (MMP-2), and MMP-9 During Dengue Virus Infection.

    Science.gov (United States)

    Her, Zhisheng; Kam, Yiu-Wing; Gan, Victor C; Lee, Bernett; Thein, Tun-Linn; Tan, Jeslin J L; Lee, Linda K; Fink, Katja; Lye, David C; Rénia, Laurent; Leo, Yee-Sin; Ng, Lisa F P

    2017-01-01

     Dengue virus infection typically causes mild dengue fever, but, in severe cases, life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) occur. The pathophysiological hallmark of DHF and DSS is plasma leakage that leads to enhanced vascular permeability, likely due to a cytokine storm.  Ninety patients with dengue during 2010-2012 in Singapore were prospectively recruited and stratified according to their disease phase, primary and secondary infection status, and disease severity, measured by plasma leakage. Clinical parameters were recorded throughout the disease progression. The levels of various immune mediators were quantified using comprehensive multiplex microbead-based immunoassays for 46 immune mediators.  Associations between clinical parameters and immune mediators were analyzed using various statistical methods. Potential immune markers, including interleukin 1 receptor antagonist, interferon γ-inducible protein 10, hepatocyte growth factor, soluble p75 tumor necrosis factor α receptor, vascular cell adhesion molecule 1, and matrix metalloproteinase 2, were significantly associated with significant plasma leakage. Secondary dengue virus infections were also shown to influence disease outcome in terms of disease severity.  This study identified several key markers for exacerbated dengue pathogenesis, notably plasma leakage. This will allow a better understanding of the molecular mechanisms of DHF and DSS in patients with dengue. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  9. Role of matrix metalloproteinase 3 in supraspinal ligament degeneration due to ankylosing spondylitis%MMP-3在强直性脊柱炎棘上韧带退变中的作用

    Institute of Scientific and Technical Information of China (English)

    许红飞; 初同伟; 张超; 张莹; 黄晨; 刘栓得

    2012-01-01

    Objective To study the expression of matrix metalloproteinase 3 (MMP-3) and its effect on supraspinal ligament degeneration due to ankylosing spondylitis ( AS ) . Methods Expression of MMP-3 and type I collagen in ligaments was detected by immunohistochemistry. Proteoglycan level in ligaments was measured by ELISA. Supraspinal ligament morphology was observed under scanning electron microscope and transmission electron microscope, respectively. Collagen fibril distribution density and positively-stained collagen area were analyzed by Image-Pro Plus(IPP). Results MMP-3 was highly expressed in vascular smooth muscle cells and endothelial cells in AS group but not in control group. The proteoglycan level was significantly higher in control group than in AS group (19. 462 ±2. 174 vs 7. 477 ± 2. 512 μg/mg). The collagen fibrils were regularly arranged in control group and apparently exposed with massive degradation of matrix in AS group. The distribution density of collagen fibrils was 218. 02 ± 10. 39/|xm2 and 122. 47 ±22. 42μm2 in control group and AS group, respectively. The proportion of positively-stained collagen area was significantly higher in control group than in AS group [ (60. 23 ±3.41) % vs (43. 37 ±5. 96)% , P <0. 05]. Conclusion MMP-3 is highly expressed in supraspinal ligament blood vessels of active AS patients by promoting the growth of blood vessels, activating the degradation of proteoglycan, and destroying the normal ultra-structure of ligaments, and is thus involved in the degenerative ossification process of ligaments.%目的 探讨基质金属蛋白酶3(matrix metalloproteinase3,MMP-3)在强直性脊柱炎(ankylosing spondylitis,AS)棘上韧带中的表达及在韧带退变中的作用.方法 免疫组化检测MMP-3、Ⅰ型胶原在韧带中的表达,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测韧带中蛋白聚糖(proteoglycan)的含量,扫描电镜及透射电镜对棘上韧

  10. Prostate hyperplasia caused by long-term obesity is characterized by high deposition of extracellular matrix and increased content of MMP-9 and VEGF.

    Science.gov (United States)

    Silva, Silas Amâncio; Gobbo, Marina Guimarães; Pinto-Fochi, Maria Etelvina; Rafacho, Alex; Taboga, Sebastião Roberto; Almeida, Eduardo Alves; Góes, Rejane Maira; Ribeiro, Daniele Lisboa

    2015-02-01

    Recent studies have shown a positive association of cancer and obesity, but the morphological and molecular mechanisms involved in this relationship are still unknown. This study analysed the impact of long-term obesity on rat prostate, focusing on stromal changes. Male adult Wistar rats were treated with high-fat diet to induce obesity, while the control group received a balanced diet. After 30 weeks of feeding, the ventral prostate was analysed by immunohistochemistry for cell proliferation, smooth muscle α-actin, vimentin, chondroitin sulphate and metalloproteinases (MMP-2 and 9). The content of androgen receptor (AR), oestrogen receptors (ERs) and vascular endothelial growth factor (VEGF) was measured by Western blotting, and activity of catalase and Glutathione-S-Transferase (GST) were quantified by enzymatic assay. Long-term obesity decreased testosterone plasma levels by 70% and resulted in stromal prostate hyperplasia, as evidenced by increased collagen fibres. Such stromal hyperplasia was associated with increased number of blood vessels and raised VEGF content, and increased expression of chondroitin sulphate, vimentin, α-actin and MMP-9. In spite of the high cell density in prostate, the proliferative activity was lower in the prostates of obese rats, indicating that hyperplasia was established during the early phases in this obesity model. AR levels increased significantly, whereas the ERα decreased in this group. Moreover, the levels of catalase and GST were changed considerably. These findings indicate that long-term obesity, besides disturbing the antioxidant control, causes intense stromal remodelling and release of factors that create an environment that can promote proliferative disorders in the gland, culminating with diffuse hyperplasia. © 2014 The Authors. International Journal of Experimental Pathology © 2014 International Journal of Experimental Pathology.

  11. Expressions of MMP-2-PEX and MMP-2 in invasive pituitary adenomas and its significance%MMP-2-PEX与MMP-2在侵袭性垂体腺瘤中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    屈洪涛; 杨伊林; 王穗暖; 刘运生; 王强; 董博; 毛宇敏

    2013-01-01

    Objective To investigate the expressions of matrix metalloproteinase-2 hemopexin domain (MMP-2-PEX) and matrix metalloproteinase-2 (MMP-2) in invasive pituitary adenomas and non-invasive pituitary adenomas, and to evaluate the relationship between the expressions of MMP-2-PEX and MMP-2 and the invasion of pituitary adenomas. Methods The expressions of MMP-2-PEX and MMP-2 mRNA were examined by reverse transcription-polymerse chain reaction assay. The relation between MMP-2-PEX and MMP-2, and the correlation between expression of MMP-2-PEX, MMP-2 and invasiveness of pituitary adenomas were studied. Results The expression of MMP-2-PEX mRNA in invasive pituitary adenomas was significantly lower than that of non-invasive pituitary adenomas (P < 0.01). The expression of MMP-2 mRNA in invasive pituitary adenomas was significantly higher than that of non-invasive pituitary adenomas (P<0.01). mRNA level of MMP-2-PEX was negatively related with mRNA of MMP-2 , and there was no correlation between them in non-invasive pituitary adenomas. Conclusion Low level of MMP-2-PEX is closely related with high level of MMP-2, so MMP-2-PEX and MMP-2 can be used as an important index for the evaluation of invasiveness of pituitary adenoma.%目的 研究基质金属蛋白酶-2在C端的血红素样结构域(MMP-2-PEX)和基质金属蛋白酶-2(MMP-2)在侵袭性垂体腺瘤中的表达及其意义.方法 用逆转录酶-聚合酶链反应(RT-PCR)法检测116例垂体腺瘤标本的MMP-2-PEX及MMP-2的表达,分析其与侵袭性垂体腺瘤的关系及两者之间的相关性.结果 侵袭性垂体腺瘤组MMP-2-PEX mRNA表达较非侵袭性垂体腺瘤组显著降低(P<0.01).侵袭性垂体腺瘤组的MMP-2 mRNA表达明显高于非侵袭性垂体腺瘤组(P<0.01).侵袭性垂体腺瘤组中MMP-2-PEX mRNA表达与MMP-2 mRNA表达呈负相关(r=-0.66,P<0.05),而非侵袭性腺垂体腺瘤组中无相关性(r =-0.22,P>0.05).结论 MMP-2-PEX低表达及MMP-2高表达与垂体腺瘤的侵袭性密切相关,MMP

  12. 溃疡性结肠炎患者粪便中Cal、MMP-9、MPO 水平检测的临床研究%Clinical studies of detection of fecal calprotectin,matrix metalloproteinase 9,myeloperoxidase in ulcerative colitis

    Institute of Scientific and Technical Information of China (English)

    朱玉; 赵孝文; 丁浩; 刘晓昌; 梅俏; 许建明

    2015-01-01

    目的:探讨溃疡性结肠炎(UC)患者粪便中钙卫蛋白(Cal)、基质金属蛋白酶9(MMP-9)、髓过氧化物酶(MPO)水平检测的临床意义。方法选择 UC 患者和正常对照者各50例,测定 UC 患者和正常对照者粪便中 Cal、MMP-9、MPO水平。结果 UC 活动期患者粪便中 Cal、MMP-9、MPO 水平显著高于缓解期患者和正常对照者,UC 患者活动期轻中重度各组粪便中 Cal、MMP-9、MPO 水平比较差异有统计学意义(P <0.05,P <0.01);UC 患者粪便中 Cal、MMP-9、MPO 水平与 DAI 评分显著相关( P <0.05)。结论粪便中 Cal、MMP-9、MPO 水平可作为 UC 患者疾病活动性评估的指标。%Objective To investigate and discuss the clinical significance of detecting the level of fecal calprotectin (Cal),matrix metalloproteinase-9( MMP-9),myelo-peroxidase( MPO)in patients with ulcerative colitis( UC). Methods To measure the level of fecal Cal,MMP-9,MPO in 50 patients with UC before and after treatment,and the level in 50 healthy controls. Results The level of fecal Cal,MMP-9,MPO in active UC was significantly high-er than the level in remission UC and healthy controls;the difference of the level of fecal Cal,MMP-9,MPO in dif-ferent clinical severity groups of mild grade,moderate grade,severe grade was statistically significant(P < 0. 05,P< 0. 01). The level of fecal Cal,MMP-9,MPO also showed significant correlation with DAI in UC(P < 0. 05). Conclusion The level of fecal Cal,MMP-9,MPO can be used as fecal makers for jurging activity in UC.

  13. Collagenase-3 (MMP-13) expression in cutaneous malignant melanoma.

    Science.gov (United States)

    Corte, M D; Gonzalez, L O; Corte, M G; Quintela, I; Pidal, I; Bongera, M; Vizoso, F

    2005-01-01

    Matrix metalloproteases (MMPs), enzymes with the ability to degrade the extracellular matrix, play an important role in tissue invasion by cutaneous malignant melanoma (CMM). One specific MMP, collagenase-3 (MMP-13), is thought to have a key function in the activation of MMP. To evaluate the expression of MMP-13 in CMM and assess its possible relationship to clinical and pathological parameters. MMP-13 expression was analyzed in 51 paraffin-embedded tumor samples from patients with invasive CMM, ten samples from in situ melanomas, and in eight samples from benign lesions (three dermal melanocytic nevi, three compound melanocytic nevi and two atypical melanocytic nevi) using immunohistochemical techniques. The median follow-up period in patients with invasive CMM was 50 months. Benign lesions were consistently negative for MMP-13, whereas three of the ten in situ melanomas (30%) and 23 of the 51 invasive CMMs (45%) showed positive immunostaining for MMP-13. The percentage of MMP-13-positive tumors correlated significantly and positively with the mitotic index (p=0.002) in invasive CMM. However, our results did not show any significant association between tumoral MMP-13 expression and relapse-free survival in patients with invasive CMM. MMP-13 appears to be a factor associated with tumor aggressiveness in CMM. It seems to eliminate an important barrier not only against tumoral invasion but also against proliferation.

  14. ERK1/2 and p38 MAP kinase control MMP-2, MT1-MMP, and TIMP action and affect cell migration : a comparison between mesothelioma and mesothelial cells

    NARCIS (Netherlands)

    Zhong, Jun; Gencay, Mikael M Cornelsen; Bubendorf, Lukas; Burgess, Janette K; Parson, Holly; Robinson, Bruce W S; Tamm, Michael; Black, Judith L; Roth, Michael

    2006-01-01

    Pleural malignant mesothelioma is a locally aggressive tumor of mesothelial cell origin. In other tumor types high expression of matrix metalloproteinase (MMP)-2, together with membrane-type1-MMP (MT1-MMP), and low levels of the tissue inhibitor of MMP (TIMP)-2 have been correlated with aggressive t

  15. Expression of MMP-2,MMP-14 and TIMP-2 in Gastric Cancer and Its Clinical Significance%MMP-2、MMP-14、TIMP-2在胃癌组织中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    张金玲; 费雁; 陈伟; 冯刚

    2013-01-01

    Objective To examine the expression of MMP-2, MMP-14 and TIMP-2 in gastric cancer and its clinical significance. Methods The expression of MMP-2,MMP-14 and TIMP-2 was immunohistochemically detected in 80 samples of gastric cancer tissues and 30 adjacent normal gastric tissues. Results The positive expression rate of MMP-2 and MMP-14 was 75.00% and 82. 50% respectively,in gastric cancer tissues,significantly higher than that in normal gastric tissues(36. 67% and 33. 33% ,respectively) (P<0. 01). The positive expression rate of TIMP-2 was 45. 00% and 40. 00% in gastric cancer tissues and normal gastric tissues with no significant difference being found. The expression of MMP-2 and MMP-14 in gastric cancer tissues was closely correlated to the cell differentiation,invasion,lymph node metastasis and TNM stages(P<0. 01). But the expression of TIMP-2 in gastric cancer tissues was only related to the lymph node metastasis (P<0. 05). The expression levels of MMP-2 and MMP-14 were significantly higher in survival time <2 years group than in survival time ≥2 years group(P<0. 01). Conclusion MMP-2 and MMP-14 are over-expressed in gastric cancer. Combined detection for MMP-2/MMP-14 or MMP-2/MMP-14/TIMP-2 may help evaluate the malignancy of gastric cancer and predict the prognosis of patients with gastric cancer.%目的 探讨基质金属蛋白酶(matrix metalloproteinase,MMP)-2、14及基质金属蛋白酶组织抑制因子-2(tissue inhibitor of metalloproteinase,TIMP-2)在胃癌中的表达及其意义.方法 用免疫组织化学方法 (SP法)检测80例胃癌手术患者胃癌组织及30例同期癌旁≥5 cm正常组织中MMP-2、MMP-14及TIMP-2的表达.结果 ①在胃癌、正常胃组织中,MMP-2阳性表达率分别为75.00%和36.67%,MMP-14阳性表达率分别为82.50%和33.33%,差异有统计学意义(均P<0.01);TIMP-2阳性表达率分别为45.00%和40.00%,差异无统计学意义.②癌组织中,MMP-2、MMP-14的表达与肿瘤分化程度、浸润深度、

  16. Role Of MMP-2 and MMP-9 in Resistance to Drug Therapy in Patients with Resistant Hypertension

    Directory of Open Access Journals (Sweden)

    Leandro Lacerda

    2015-01-01

    Full Text Available Background: Despite the increased evidence of the important role of matrix metalloproteinases (MMP-9 and MMP‑2 in the pathophysiology of hypertension, the profile of these molecules in resistant hypertension (RHTN remains unknown. Objectives: To compare the plasma levels of MMP-9 and MMP-2 and of their tissue inhibitors (TIMP-1 and TIMP-2, respectively, as well as their MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios, between patients with controlled RHTN (CRHTN, n=41 and uncontrolled RHTN (UCRHTN, n=35. In addition, the association of those parameters with clinical characteristics, office blood pressure (BP and arterial stiffness (determined by pulse wave velocity was evaluate in those subgroups. Methods: This study included 76 individuals diagnosed with RHTN and submitted to physical examination, electrocardiogram, and laboratory tests to assess biochemical parameters. Results: Similar values of MMP-9, MMP-2, TIMP-1, TIMP-2, and MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios were found in the UCRHTN and CRHTN subgroups (P>0.05. A significant correlation was found between diastolic BP (DBP and MMP-9/TIMP-1 ratio (r=0.37; P=0.02 and DPB and MMP-2 (r=-0.40; P=0.02 in the UCRHTN subgroup. On the other hand, no correlation was observed in the CRHTN subgroup. Logistic regression models demonstrated that MMP-9, MMP-2, TIMP-1, TIMP-2 and their ratios were not associated with the lack of BP control. Conclusion: These findings suggest that neither MMP-2 nor MMP-9 affect BP control in RHTN subjects.

  17. Expression of GPC3, MMP-9 and MMP-14 in Hepatocellular Carcinoma and Their Influence on the Prognosis of Patients

    Directory of Open Access Journals (Sweden)

    Lei CAI

    2016-03-01

    Full Text Available Objective: To explore the expression of glypican-3 (GPC3, metal matrix proteinase (MMP-9 Methods: Totally 112 paraffin-embedded tissue samples of HCC patients were selected as observation group and 70 normal tissue samples were as control group. The expressions of GPC3, MMP-9 and MMP-14 of two groups were detected using immunohistochemistry assay. The positive rates of two groups were calculated. The relationship between the expression of GPC3, MMP-9 and MMP-14 and clinicopathological features, and their influence on the survival time of HCC patients were compared. Results: The positive expression rates of GPC3, MMP-9 and MMP-14 were higher in observation group that those in control group, the differences were statistically significant (P=0.000; P=0.000; P=0.000. The expression of GPC3 had close relationship with tumor volume, differentiated degree, lymphatic metastasis, and PCNA expression. The expression of MMP-9 had close relationship with tumor volume, lymphatic metastasis, and vascular invasion. The expression of GPC3 had close relationship with tumor volume, differentiated degree, lymphatic metastasis, vascular invasion, and proliferating cell nuclear antigen (PCNA expression.Conclusion: GPC3, MMP-9 and MMP-14 are highly expressed in HCC patients, which shows poor prognosis. Therefore, the detection of GPC3, MMP-9 and MMP-14 after surgery has a certain value on assessment of the prognosis of HCC patients.and MMP-14 in hepatocellular carcinoma (HCC, and their influence on the prognosis of HCC patients. There were positive correlations between GPC3 and MMP-9 (r=0.538, P=0.042, MMP-9 and MMP-14 (r=0.430, P=0.024, and GPC3 and MMP-14Kaplan-Meier method showed that the expressions of GPC3, MMP-9 and MMP-14 were associated with the prognosis of HCC patients, and patients with high expressions of GPC3, MMP-9 and MMP-14 had poor prognosis. (r=0.563, P=0.563.

  18. An ex vivo study on immunohistochemical localization of MMP-7 and MMP-9 in temporomandibular joint discs with internal derangement

    Directory of Open Access Journals (Sweden)

    C. Loreto

    2013-04-01

    Full Text Available Internal derangement (ID is among the most common disorders of the temporomandibular joint (TMJ. Previous research by our group highlighted a correlation between apoptosis and TMJ ID. Metalloproteinases (MMP-7 and -9 have been shown to play an important role in extracellular matrix ECM homeostasis and, through it, in joint disc remodelling. The immunohistochemical expression of MMP-7 and -9 was investigated in discs from patients with TMJ ID and from healthy donors and compared with the degree of histological tissue degeneration. The collagen fibre arrangement in pathological discs exhibited varying degrees of disruption. New vessels were consistently detected; endothelial cells from these vessels were immunolabelled with both MMP-7 and MMP-9. More or less intense MMP-7 and MMP-9 immunolabelling was detected in the cytoplasm of disc cells from all patients. MMP-7 and MMP-9 immunostaining was significantly different between pathological and normal discs and correlated with the extent of histopathological degeneration. MMP-7 and MMP-9 upregulation in discs from patients with TMJ ID demonstrates their involvement in disc damage in this disorder. A greater understanding of these processes could help identify ways to curb MMP overproduction without affecting their tissue remodelling action. The design of specific inhibitors for these MMPs would not only help to gain insights into the biological roles of MMPs, but would also aid in developing therapeutic interventions for diseases associated with abnormal ECM degradation.

  19. Effect of LED irradiation on the expression of MMP-3 and MMP-13 in SW1353 cells in vitro

    Science.gov (United States)

    Zeng, Chang-chun; Guo, Zhou-yi; Zhang, Feng-xue; Deng, Wen-di; Liu, Song-hao

    2007-05-01

    Matrix Metalloproteinase (MMP) plays an active role in remodeling cartilage in osteoarthritic cartilage. To find an effective method of prevention of osteoclasia, this in vitro study focuses on the expression of MMP-3 and MMP-13 in the SW1353 cells by LED irradiation. The human chondrosarcoma cell line SW1353 were stimulated with the proinflammatory cytokine IL-1beta or tumor necrosis factor-alpha (TNF-alpha), and were received the irradiation of LED (632nm, 4mW/cm2). The cell count was assessed over a 96-hour period by using Trypan blue dye exclusion assay, and the cell activity was evaluated with a Cell Counting Kit-8 Assays. The subsequent expression of MMP-3 and MMP-13 was quantified. Results of this experiment showed that the cultural cell activity was decreased, and the expression of MMP-3 and MMP-13 was increased by being stimulated with IL-1beta or TNF-alpha. After received LED irradiation, the death rate of cultural cell was increased and the expression of MMP-3 and MMP-13 was decreased significantly. The present study concluded that particular LED irradiation stimulates SW1353 cell proliferation activity and inhibit the MMP-3 and MMP-13 enzymatic activity. These findings might be clinically relevant, indicating that the low power laser irradiation treatment is likely to achieve the repair of articular cartilage in clinic.

  20. Stomach Cancer: Interconnection between the Redox State, Activity of MMP-2, MMP-9 and Stage of Tumor Growth.

    Science.gov (United States)

    Burlaka, Anatoly P; Ganusevich, Irina I; Gafurov, Marat R; Lukin, Sergey M; Sidorik, Evgeny P

    2016-04-01

    High levels of reactive oxygen (ROS) and nitrogen (RNS) species can lead to the destruction of extracellular matrix facilitating tumor progression. ROS can activate matrix metalloproteinases (MMP), damage DNA and RNA. Therefore, the levels of MMP, ROS and RNS can serve as additional prognostic markers and for the estimation of the effectiveness of tumor therapy. Concerning gastric cancer, the prognostic role of MMP, its connection with the cancer staging remains controversial and correlations between the activity of MMP with the ROS and RNS levels are insufficiently confirmed. Superoxide generation rates, nitric oxide (NO) levels, concentrations of active forms of matrix metalloproteinases MMP-2 and MMP-9 in tumor and adjacent tissues of patients with stomach cancer at different disease stages were measured by electron spin resonance (ESR) including spin-trapping and polyacrylamide gel zymography. It is shown that the activity of MMP-2 and MMP-9 in tumor tissue correlate with the superoxide radicals generation rate and NO levels (r = 0.48÷0.67, p < 0.05). The activity of MMP-2 and MMP-9 in tumor tissues and superoxide radical generation rates correlate positively with the stage of regional dissemination (r = 0.45 and 0.37, correspondingly, p < 0.05), but MMP-2 and MMP-9 activity inversely depends on distant metastatic degree of stomach cancer (r = 0.58; p < 0.05). Additionally, the feasibility of ESR to locally determine oxidative stress is demonstrated.

  1. Expression of MMP-2, MT1-MMP, and TIMP-2 by cultured rabbit corneal fibroblasts under mechanical stretch.

    Science.gov (United States)

    Liu, Chengxing; Feng, Pengfei; Li, Xiaona; Song, Jie; Chen, Weiyi

    2014-08-01

    Refractive surgery not only leads to tissue injury but also evokes mechanical stress increase of the cornea. How the mechanical stress affects the corneal matrix remodeling, specifically, matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of metalloproteinases; TIMPs) is not well understood. In this study, cultured rabbit corneal fibroblasts in vitro were subjected to regimen of 5%, 10%, or 15% equibiaxial stretch at 0.1 Hz for 3 or 24 h. MMP-2 protein level was measured by gelatin zymography and Western blotting. MMP-2, membrane type 1 MMP (MT1-MMP), and TIMP-2 mRNA levels were quantified by real-time quantitative PCR. Extracellular regulated protein kinase (ERK) phosphorylation protein levels were assessed by Western blotting. Our results showed that a 15% stretch resulted in increases in MMP-2 protein, MMP-2 mRNA, and MT1-MMP mRNA levels, but a decrease in TIMP-2 mRNA level. However, a 5% stretch caused decreases in MMP-2 protein and mRNA level, but an increase in TIMP-2 mRNA level, and no change in MT1-MMP mRNA level. A 15% stretch also caused a significant increase in ERK1/2 phosphorylation. Inhibition of the mitogenactivated protein kinase (MEK) pathway with PD98059 attenuated stretch-induced increase in MMP-2 production and ERK activity. These results suggest that small-magnitude stretching may promote corneal matrix synthetic events, whereas large-magnitude stretching promotes corneal matrix degradation by changing the balance between MMPs and TIMPs in corneal fibroblasts. Large-magnitude stretch-induced increase in pro-MMP-2 production was in an ERK-dependent manner. © 2014 by the Society for Experimental Biology and Medicine.

  2. Macrophage migration and invasion is regulated by MMP10 expression.

    Directory of Open Access Journals (Sweden)

    Megan Y Murray

    Full Text Available This study was designed to identify metalloproteinase determinants of macrophage migration and led to the specific hypothesis that matrix metalloproteinase 10 (MMP10/stromelysin-2 facilitates macrophage migration. We first profiled expression of all MMPs in LPS-stimulated primary murine bone marrow-derived macrophages and Raw264.7 cells and found that MMP10 was stimulated early (3 h and down-regulated later (24 h. Based on this pattern of expression, we speculated that MMP10 plays a role in macrophage responses, such as migration. Indeed, using time lapse microscopy, we found that RNAi silencing of MMP10 in primary macrophages resulted in markedly reduced migration, which was reversed with exogenous active MMP10 protein. Mmp10 (-/- bone marrow-derived macrophages displayed significantly reduced migration over a two-dimensional fibronectin matrix. Invasion of primary wild-type macrophages into Matrigel supplemented with fibronectin was also markedly impaired in Mmp10 (-/- cells. MMP10 expression in macrophages thus emerges as an important moderator of cell migration and invasion. These findings support the hypothesis that MMP10 promotes macrophage movement and may have implications in understanding the control of macrophages in several pathologies, including the abnormal wound healing response associated with pro-inflammatory conditions.

  3. Gelatinase B/MMP-9 in Tumour Pathogenesis and Progression

    Energy Technology Data Exchange (ETDEWEB)

    Farina, Antonietta Rosella; Mackay, Andrew Reay, E-mail: andrewreay.mackay@univaq.it [Department of Applied Clinical and Biotechnological Sciences, University of L’Aquila, Via Vetoio, Coppito 2, L’Aquila 67100 (Italy)

    2014-01-27

    Since its original identification as a leukocyte gelatinase/type V collagenase and tumour type IV collagenase, gelatinase B/matrix metalloproteinase (MMP)-9 is now recognised as playing a central role in many aspects of tumour progression. In this review, we relate current concepts concerning the many ways in which gelatinase B/MMP-9 influences tumour biology. Following a brief outline of the gelatinase B/MMP-9 gene and protein, we analyse the role(s) of gelatinase B/MMP-9 in different phases of the tumorigenic process, and compare the importance of gelatinase B/MMP-9 source in the carcinogenic process. What becomes apparent is the importance of inflammatory cell-derived gelatinase B/MMP-9 in tumour promotion, early progression and triggering of the “angiogenic switch”, the integral relationship between inflammatory, stromal and tumour components with respect to gelatinase B/MMP-9 production and activation, and the fundamental role for gelatinase B/MMP-9 in the formation and maintenance of tumour stem cell and metastatic niches. It is also apparent that gelatinase B/MMP-9 plays important tumour suppressing functions, producing endogenous angiogenesis inhibitors, promoting inflammatory anti-tumour activity, and inducing apoptosis. The fundamental roles of gelatinase B/MMP-9 in cancer biology underpins the need for specific therapeutic inhibitors of gelatinase B/MMP-9 function, the use of which must take into account and substitute for tumour-suppressing gelatinase B/MMP-9 activity and also limit inhibition of physiological gelatinase B/MMP-9 function.

  4. Correlation Between Th1, Th2 Cells and Levels of Serum MMP-2, MMP-9 in Children with Asthma

    Directory of Open Access Journals (Sweden)

    Xuan WANG

    2015-12-01

    Full Text Available Abstract Objective: To explore the correlation between Th1 and Th2 cells and the levels of serum matrix metalloproteinase-2 (MMP-2 and MMP-9 in children with asthma. Methods: A total of 89 children with asthma were divided into acute group (n=48 and chronic group (n=41 according to the course of disease, and 40 healthy children at the same term were collected as control group. The ratios of Th1 and Th2 cells as well as levels of MMP-2 and MMP-9 were compared in three groups, and the correlation between Th1 and Th2 cells and levels of MMP-2, MMP-9 was analyzed in acute group and chronic group. Results: When compared with control group, the ratios of Th1 and Th2 cells went down in both acute group and chronic group (P<0.01, while the levels of serum MMP-2 and MMP-9 up (P<0.01. The levels of serum MMP-2 and MMP-9 in acute group were dramatically higher than those in chronic group, and there was statistical significance (P<0.01. Pearson correlation analysis revealed that there was no significant correlation between Th1 and Th2 cells and MMP-2 level (r=0.148, P=0.314, r=0.299, P=0.058; r=0.183, P=0.214, r=0.289, P=0.067, whereas both Th1 and Th2 cells were negatively correlated with MMP-9 level in acute group and chronic group (r=-0.489, P=0.000, r=-0.324, P=0.039; r=-0.352, P=0.014, r=-0.357, P=0.022. Conclusion: Aberrant secretion of Th cells can not only damage the immune function of children with asthma, but also decrease the level of serum MMP-9, consequently affecting the collagen degradation and airway remodeling.

  5. MMP-10 is required for efficient muscle regeneration in mouse models of injury and muscular dystrophy.

    Science.gov (United States)

    Bobadilla, Míriam; Sáinz, Neira; Rodriguez, José Antonio; Abizanda, Gloria; Orbe, Josune; de Martino, Alba; García Verdugo, José Manuel; Páramo, José A; Prósper, Felipe; Pérez-Ruiz, Ana

    2014-02-01

    Matrix metalloproteinases (MMPs), a family of endopeptidases that are involved in the degradation of extracellular matrix components, have been implicated in skeletal muscle regeneration. Among the MMPs, MMP-2 and MMP-9 are upregulated in Duchenne muscular dystrophy (DMD), a fatal X-linked muscle disorder. However, inhibition or overexpression of specific MMPs in a mouse model of DMD (mdx) has yielded mixed results regarding disease progression, depending on the MMP studied. Here, we have examined the role of MMP-10 in muscle regeneration during injury and muscular dystrophy. We found that skeletal muscle increases MMP-10 protein expression in response to damage (notexin) or disease (mdx mice), suggesting its role in muscle regeneration. In addition, we found that MMP-10-deficient muscles displayed impaired recruitment of endothelial cells, reduced levels of extracellular matrix proteins, diminished collagen deposition, and decreased fiber size, which collectively contributed to delayed muscle regeneration after injury. Also, MMP-10 knockout in mdx mice led to a deteriorated dystrophic phenotype. Moreover, MMP-10 mRNA silencing in injured muscles (wild-type and mdx) reduced muscle regeneration, while addition of recombinant human MMP-10 accelerated muscle repair, suggesting that MMP-10 is required for efficient muscle regeneration. Furthermore, our data suggest that MMP-10-mediated muscle repair is associated with VEGF/Akt signaling. Thus, our findings indicate that MMP-10 is critical for skeletal muscle maintenance and regeneration during injury and disease.

  6. MMP-9 and MMP-2 gelatinases and TIMP-1 and TIMP-2 inhibitors in breast cancer: correlations with prognostic factors.

    Science.gov (United States)

    Jinga, D C; Blidaru, A; Condrea, Ileana; Ardeleanu, Carmen; Dragomir, Cristina; Szegli, G; Stefanescu, Maria; Matache, Cristiana

    2006-01-01

    The goal of our study was to analyse the prognostic values for some matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) in breast cancer. We evaluated the activity and the expression levels of MMP-9, MMP-2, TIMP-1 and TIMP-2 in malignant versus benign fresh breast tumor extracts. For this purpose, gelatinzymography, immunoblotting and ELISA were used to analyse the activity and expression of MMPs and TIMPs. We found that MMP-9 expression level and activity are increased in malignant tumors. In addition, MMP-9/TIMP-1 and MMP-2/TIMP-2 ratio values obtained by us were significantly different in malignant tumors compared to benign tumors. We suggest that the abnormal MMP-9/TIMP-1 balance plays a role in the configuration of breast invasive carcinoma of no special type and also in tumor growth, while altered MMP-2/TIMP-2 ratio value could be associated with lymph node invasion and used as a prognostic marker in correlation with Nottingham Prognostic Index. Finally, we showed that in malignant tumors high expression of estrogen receptors is associated with enhanced activity of MMP-2 and increased bcl- 2 levels, while high expression of progesterone receptors is correlated with low TIMP-1 protein levels.

  7. Effect of simulated high altitude hypoxia on matrix metalloproteinase-2 expression in serum of rats with chronic periodontitis%模拟高原缺氧对慢性牙周炎大鼠血清中MMP-2表达的影响

    Institute of Scientific and Technical Information of China (English)

    孔耀; 刘鲁川; 周霞; 刘福玉; 杨苏平; 张海元

    2010-01-01

    目的 研究常氧与缺氧条件下慢性牙周炎大鼠血清中基质金属蛋白酶2(matrix metalloproteinase-2, MMP-2)的表达变化,探讨MMP-2与高原牙周病的关系.方法 模拟高原缺氧建立大鼠慢性牙周炎模型,采用ELISA法测定正常对照组、常氧牙周炎组、缺氧对照组、缺氧牙周炎组大鼠血清中MMP-2表达.结果 正常对照组、常氧牙周炎组、缺氧对照组、缺氧牙周炎组血清中MMP-2浓度分别为(92.00±21.78)、(118.09±25.01)、(102.03±21.30)、(159.19±34.13) ng/ml,常氧牙周炎组与正常对照组、缺氧牙周炎组与缺氧对照组、缺氧对照组与正常对照组、缺氧牙周炎组与常氧牙周炎组比较均有统计学意义(P<0.01).其中缺氧牙周炎组血清中MMP-2浓度升高最为显著(P<0.01).结论 缺氧上调血清中MMP-2的表达,进而促进大鼠牙周组织的破坏.

  8. MMP-1 and MMP-9 regulate epidermal growth factor-dependent collagen loss in human carotid plaque smooth muscle cells.

    Science.gov (United States)

    Rao, Velidi H; Kansal, Vikash; Stoupa, Samantha; Agrawal, Devendra K

    2014-02-01

    Mechanisms underlying the rupture of atherosclerotic plaque, a crucial factor in the development of myocardial infarction and stroke, are not well defined. Here, we examined the role of epidermal growth factor (EGF)-mediated matrix metalloproteinases (MMP) on the stability of interstitial collagens in vascular smooth muscle cells (VSMCs) isolated from carotid endarterectomy tissues of symptomatic and asymptomatic patients with carotid stenosis. VSMCs isolated from the carotid plaques of both asymptomatic and symptomatic patients were treated with EGF. The MMP-9 activity was quantified by gelatin zymography and the analysis of mRNA transcripts and protein for MMP-9, MMP-1, EGFR and collagen types I, Col I(α1) and collagen type III, Col III(α1) were analyzed by qPCR and immunofluorescence, respectively. The effect of EGF treatment to increase MMP-9 activity and mRNA transcripts for MMP-9, MMP-1, and EGFR and to decrease mRNA transcripts for Col I(α1) and Col III(α1) was threefold to fourfold greater in VSMCs isolated from the carotid plaques of symptomatic than asymptomatic patients. Inhibitors of EGFR (AG1478) and a small molecule inhibitor of MMP-9 decreased the MMP9 expression and upregulated Col I(α1) and Col III(α1) in EGF-treated VSMCs of both groups. Additionally, the magnitude in decreased MMP-9 mRNA and increased Col I(α1) and Col III(α1) due to knockdown of MMP-9 gene with siRNA in EGF-treated VSMCs was significantly greater in the symptomatic group than the asymptomatic group. Thus, a selective blockade of both EGFR and MMP-9 may be a novel strategy and a promising target for stabilizing vulnerable plaques in patients with carotid stenosis.

  9. Ginseng and Its Active Components Ginsenosides Inhibit Adipogenesis in 3T3-L1 Cells by Regulating MMP-2 and MMP-9

    Directory of Open Access Journals (Sweden)

    Jaeho Oh

    2012-01-01

    Full Text Available The growth and development of adipose tissue are believed to require adipogenesis, angiogenesis, and extracellular matrix remodeling. As our previous study revealed that ginseng reduces adipose tissue mass in part by decreasing matrix metalloproteinase (MMP activity in obese mice, we hypothesized that adipogenesis can be inhibited by ginseng and its active components ginsenosides (GSs. Treatment of 3T3-L1 adipocytes with Korean red ginseng extract (GE inhibited lipid accumulation and the expression of adipocyte-specific genes (PPARγ, C/EBPα, aP2, and leptin. GE decreased both the mRNA levels and activity of MMP-2 and MMP-9 in 3T3-L1 cells. These effects were further inhibited by total GSs (TGSs and individual GSs. TGSs and individual GSs also significantly decreased MMP-2 and MMP-9 reporter gene activities in the presence of phorbol 12-myristate 13-acetate (PMA, the MMP inducer. Among the GSs, Rb1 most effectively inhibited MMP activity. In addition, PMA treatment attenuated the inhibitory actions of GE and GSs on adipogenesis. Moreover, GE and GSs reduced the expression of NF-κB and AP-1, the transcription factors of MMP-2 and MMP-9. These results demonstrate that ginseng, in particular GSs, effectively inhibits adipogenesis and that this process may be mediated in part through the suppression of MMP-2 and MMP-9. Thus, ginseng and GSs likely have therapeutic potential for controlling adipogenesis.

  10. Occurrence of Two Distinct Types of Tissue Inhibitors of Metallo-proteinases-2 in Fugu rubripes

    Institute of Scientific and Technical Information of China (English)

    Yoshihiro Yokoyama; Hiroshi Tsukamoto; Tohru Suzuki; Shohshi Mizuta; Reiji Yoshinaka

    2005-01-01

    In this study, genes of two distinct tissue inhibitors of metalloproteinases-2 (TIMP-2) from Japanese puffer fish Fugu rubripes, Fugu TIMP-2a and TIMP-2b, were cloned. The open reading frames of Fugu TIMP-2a and TIMP-2b cDNAs are composed of 660 and 657 nucleotides and 220 and 219 amino acids, respectively. Both Fugu TIMP-2s contain 12 cysteine residues, whichmight form six disulfide bonds as in other animals TIMP-2s. Reverse-transcribed polymerase chain reaction analysis showed the mRNAs of Fugu TIMP-2a and TIMP-2b to be expressed in some tissues examined with different expression patterns. These findings suggest that the two distinct Fugu TIMP-2s might perform different functions in Fugu tissues.

  11. Pre-Treatment of Platinum Resistant Ovarian Cancer Cells with an MMP-9/MMP-2 Inhibitor Prior to Cisplatin Enhances Cytotoxicity as Determined by High Content Screening

    Directory of Open Access Journals (Sweden)

    John J. O'Leary

    2013-01-01

    Full Text Available Platinum resistance is a major cause of treatment failure in ovarian cancer. We previously identified matrix metalloproteinase 9 (MMP-9 as a potential therapeutic target of chemoresistant disease. A2780cis (cisplatin-resistant and A2780 (cisplatin-sensitive ovarian carcinoma cell lines were used. The cytotoxic effect of MMP-9/MMP-2 inhibitor, (2R-2-[(4-Biphenylsulfonyl amino]-3 phenylpropionic acid (C21H19NO4S alone or in combination with cisplatin was determined using high content screening. Protein expression was examined using immunohistochemistry and ELISA. Co-incubation of cisplatin and an MMP-9/MMP-2 inhibitor, (2R-2-[(4-Biphenylsulfonyl amino]-3 phenylpropionic acid (C21H19NO4S resulted in significantly greater cytotoxicity as compared to either treatment alone in a cisplatin resistant MMP-9 overexpressing cell line; A2780cis. In addition, pre-incubating with MMP-9i prior to cisplatin further enhances the cytotoxic effect. No significant difference was observed in MMP-9 protein in tissue but a trend towards increased MMP-9 was observed in recurrent serum. We propose that MMP-9/MMP-2i may be utilized in the treatment of recurrent/chemoresistant ovarian cancers that overexpress MMP-9 mRNA but its role in vivo remains to be evaluated.

  12. In vitro modulation of MMP-2 and MMP-9 in human cervical and ovarian cancer cell lines by cytokines, inducers and inhibitors.

    Science.gov (United States)

    Roomi, M W; Monterrey, J C; Kalinovsky, T; Rath, M; Niedzwiecki, A

    2010-03-01

    Matrix metalloproteinases (MMPs) secreted by cervical and ovarian cancer, especially MMP-2 and MMP-9, play crucial roles in tumor invasion and metastasis. We examined the effect of cytokines, mitogens, inducers and inhibitors on MMP-2 and MMP-9 expression in cervical and ovarian cancer cell lines. Human cervical (HeLa and DoTc2-4510) and ovarian (SK-OV-3) cell lines were cultured in appropriate media. At near confluence, the cells were washed with PBS and incubated in serum-free medium with various concentrations of several cytokines, mitogens and inhibitors. After 24 h the media were removed and analyzed for MMP-2 and MMP-9 by gelatinase zymography and quantitated by densitometry. HeLa and SK-OV-3 cell lines expressed MMP-2 whereas DoTc2-4510 cells expressed MMP-9. Treatment of cervical cancer cell lines (HeLa and DoTc2-4510) with PMA had no effect on MMP-2 expression and a moderate stimulatory effect in ovarian cancer cell line SK-OV-3. MMP-9 was stimulated by phorbol 12-myristate 13-acetate in HeLa cells and enhanced in DoTc2-4510. Tumor necrosis factor-alpha and interleukin-1beta, had slight inhibitory effect on HeLa cell expression of MMP-2 while lipopolysaccharide stimulated MMP-2 in HeLa cells. Doxycycline, epigallocatechin gallate, a nutrient mixture, actinomycin-D, cyclohexamide, retinoic acid and dexamethasone inhibited MMP-2 in HeLa and SK-OV-3 cell lines and inhibited MMP-9 in DoTc2-4510. Our results show that cytokines, mitogens, inducers and inhibitors have an up or down regulatory effect on MMP-2 and MMP-9 expression in ovarian and cervical cancer cell lines, suggesting these agents may be effective strategies to treat these cancers.

  13. The expressions and their significances of CD147,matrix metalloproteinase-9 and vascular endothelial growth factor in renal cell carcinoma%CD147、MMP-9与VEGF在肾细胞癌中的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    王苏春; 王禾; 秦卫军; 孙航; 顾汝军

    2011-01-01

    Objective:To study the expressions of CD147 ,Matrix metalloproteinase-9( MMP-9) and vascular endothelial growth factor ( VEGF) in renal cell carcinoma( RCC) ,and explore their correlations with tumor growth,tumor invasion and metastasis. Methods:The expressions of CD147,MMP-9 and VEGF were detected by immunohistochemical SP method in tissue from the carcinoma( RCC,n = 107) and normal renal tissue ( n = 10). The relationships between the expressions and clinical pathology were analyzed, and the correlations among CD147, MMP-9 and VEGF were studied. Results: The expressions of CD147, MMP-9 and VEGF in RCC were significantly higher than those in normal renal tissue (P 0.05) ,but had statistical significances related to clinical staging(P 0. 05 ) . Conclusions: CD147, MMP-9 and VEGF play the important role in the progression and invasion in RCC,the detection of CD147,MMP-9 and VEGF expressions may be the potential biological indexes which reflect tumor growth, invasion and metastasis of RCC.%目的:探讨白细胞分化抗原147(CD147)、基质金属蛋白酶-9(MMP-9)与血管内皮生长因子(VEGF)在肾细胞癌(RCC)组织中的表达及其与肾癌组织发生、肿瘤侵袭和转移的关系.方法:采用免疫组织化学SP法,检测107例RCC组织、10例正常肾组织中CD147、MMP-9与VEGF的表达情况,分析其表达与RCC的临床分期、病理分级的关系及其三者间的相关性.结果:RCC中CD147、MMP-9与VEGF表达均明显高于正常肾组织(P0.05),不同临床分期RCC组织CD147、MMP-9与VEGF表达差异均有统计学意义(P0.05),在肿瘤大小、病理分型、临床分期及肿瘤转移间差异均有统计学意义(P<0.01).结论:CD147、MMP-9与VEGF在RCC的临床演进中起重要作用.检测CD147、MMP-9与VEGF可作为反映RCC发生及侵袭转移潜能的生物学指标.

  14. Characterization of porcine MMP-2 and its association with immune traits

    DEFF Research Database (Denmark)

    Huang, Honggang; Zhao, Weimin; Tang, Zhonglin;

    2009-01-01

    Matrix metalloproteinase-2 (MMP-2) plays important roles in inflammation and immunity besides its basic role in degrading and remodelling extracellular matrix (ECM). The expression of MMP-2 is up-regulated in many human as well as animal models of inflammatory and immune diseases. In this study, we...

  15. Elevation of MMP-3 and MMP-9 in CSF and Blood in Patients with Severe Traumatic Brain Injury

    Science.gov (United States)

    Grossetete, Mark; Phelps, Jeremy; Arko, Leopold; Yonas, Howard; Rosenberg, Gary A.

    2009-01-01

    OBJECTIVE Traumatic brain injury (TBI) causes elevation of matrix metalloproteinases (MMPs), which are associated with neuroinflammation, blood-brain barrier (BBB) disruption, hemorrhage and cell death. We hypothesized that patients with TBI have an increase in MMPs in the ventricular cerebrospinal fluid (CSF) and plasma. METHODS Patients with TBI and a ventricular catheter were entered into the study. Samples of CSF and plasma were collected at the time of catheter placement, and 24 and 72 hrs after admission. Seven TBI patients were entered into the study with six having complete data for analysis. Only patients that had a known time of insult that fell within a six hour window from initial insult to ventriculostomy were accepted into the study. Control CSF came from ventricular fluid in patients undergoing shunt placement for normal pressure hydrocephalus (NPH). Both MMP-2 and MMP-9 were measured with gelatin zymography and MMP-3 with Western immunoblotting. RESULTS We found a significant elevation in the levels of the latent form of MMP-9 (92-kDa) in the CSF obtained at the time of arrival (TOA) (p<0.05). Elevated levels of MMP-2 were detected in plasma at 72 hours, but not in the CSF. Using albumin from both CSF and blood, we calculated the MMP-9 index, which was significantly elevated in the CSF, indicating endogenous MMP production. Western immunoblots showed increased levels of MMP-3 in CSF at all times measured, while MMP-3 was not detected in the CSF of NPH. CONCLUSIONS We show that MMPs are elevated in CSF of TBI patients. Although the number of patients was small, the results were robust and clearly demonstrated elevations of MMP-3 and MMP-9 in ventricular CSF in TBI patients compared to controls. While these preliminary results will need to be replicated, we propose that MMPs may be important in BBB opening and hemorrhage secondary to brain injury in patients. PMID:19834375

  16. Adenovirus-mediated Gene Transfer of MMP-2 into Cultured Porcine Trabecular Meshwork Cells

    OpenAIRE

    2012-01-01

    This study aimed to use adenoviral gene transfer to express matrix metalloproteinase (MMP)-2 in cultured porcine trabecular meshwork cells and to evaluate the duration of adenovirus-mediated MMP-2 expression and its enzymatic activity. MMP-2 cDNA was synthesized by ligating three segments of MMP-2 cDNA obtained by reverse transcription-polymerase chain reaction (RT-PCR) with mRNA extracted from mouse lungs. MMP-2 cDNA was inserted into replication-deficient adenoviral vectors. Western blottin...

  17. AGEs对兔软骨细胞TNF-α和MMP-13表达的影响及其机制研究%The effects of advanced glycation end products on expression of tumor necrosis factor-αand matrix metalloproteinase-13 in rabbit chondrocytes and its mechanism

    Institute of Scientific and Technical Information of China (English)

    陈铖; 马翅; 张莹; 肖钧; 蔡巍; 谭海涛

    2013-01-01

    目的:探讨晚期糖基化终末产物(AGEs)对兔软骨细胞肿瘤坏死因子-α(TNF-α)和基质金属蛋白酶-13(MMP-13)的影响及可能机制。方法:不同浓度的AGEs与兔软骨细胞共孵育48h后采用RT-PCR方法检测TNF-α和MMP-13的mRNA表达量,试剂盒方法检测过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)水平,荧光探针法检测细胞内活性氧(ROS)水平。AGEs与兔软骨细胞共孵育的同时,分别加入AGEs受体的抗体(Anti-RAGE)及核因子-κB(NF-κB)的特异性阻断剂PDTC处理,同法检测TNF-α和MMP-13的mRNA表达量。结果:与AGEs共孵育48h后兔软骨细胞TNF-α及MMP-13表达明显增多,CAT、SOD活性降低,MDA、ROS含量增多,均呈浓度依赖性;分别加入Anti-RAGE 及PDTC 处理后软骨细胞TNF-α及MMP-13表达明显低于AGEs单独处理组(P0.05)。结论:AGEs能诱导软骨细胞TNF-α和MMP-13表达增多,其机制与激活RAGE,诱导ROS生成增多,激活NF-κB信号通路有关。%Objective To detect the effects of advanced glycation end products (AGEs) on expression of tu-mor necrosis factor-α(TNF-α) and matrix metalloproteinase-13(MMP-13) in rabbit chondrocytes and its mecha-nism. Methods The chondrocytes were incubated with different concentrations of AGEs for 48h, the expression of TNF-αand MMP-13 mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR),the lev-el of catalase (CAT), Malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were detected by kits, and the level of ROS is measured by the method of Fluorescent probe. The effect of anti-RAGE(Receptor for AGEs) and PDTC(the inhibitor of NF-KB) on the expression of TNF-αand MMP-13 induced by AGEs were also measured by RT-PCR. Results After chondrocytes were incubated with AGEs, dose-dependly increased the ex-pression of TNF-αand MMP-13 (P0.05). Conclusion AGEs can induce the TNF-αand MMP-13 expression. It's mechanism may

  18. Effects of atorvastatin on the expression levels of tissue inhibitor of metalloproteinases timps/matrix metalloproteinases in bone tissues of femoral head necrosis induced by glucocorticoid%阿托伐他汀对糖皮质激素性股骨头坏死骨组织MMP/TIMP系统的影响

    Institute of Scientific and Technical Information of China (English)

    王建忠; 武永刚; 董慧珍

    2014-01-01

    目的:观察阿托伐他汀对长期应用皮质激素大鼠骨组织中基质金属蛋白酶-2(matrixmetallo-proteinase-2,MMP-2)、基质金属蛋白酶-9( matrix metalloproteinase-9,MMP-9)及其特异性抑制因子基质金属蛋白酶组织抑制剂-1( tissue inhibitor of matrix metalloproteinases-1,TIMP-1)和基质金属蛋白酶组织抑制剂-2( tissue inhibitor of matrix metalloproteinases-2,TIMP-2) mRNA表达的影响,探讨阿托伐他汀预防激素性股骨头坏死的效果及其作用机制。方法健康SD大鼠30只,采用数字随机法,分为激素组、阿托伐他汀组和对照组3个组,每组10只。激素组和阿托伐他汀组给予肌内注射醋酸泼尼松龙12.5 mg/kg,每周2次;阿托伐他汀组同时给予阿托伐他汀1 mg/kg灌胃,每日1次(按每千克体重最大用量给药,每天最大用量是60 mg,实验动物给药量为60 mg/60 kg体重);对照组只给予相同体积生理盐水肌注。给药后4周取左侧股骨头骨组织石蜡包埋,HE 染色,鉴定骨质疏松和股骨头坏死情况;取右侧股骨头骨组织提取总 RNA,采用逆转录聚合酶链反应(RT-PCR)技术检测MMP-2、MMP-9、TIMP-1和TIMP-2的mRNA表达水平。结果激素组与阿托伐他汀组各有1只动物死亡。对照组股骨头骨组织切片HE染色可见骨小梁由板层骨构成,绝大多数小梁骨陷窝内可见骨细胞,小梁间为血管和骨髓。激素组表现为骨小梁稀疏和大量不连续的骨碎片及骨髓坏死,碎片骨陷窝内骨细胞大部分消失,周围有大量炎性肉芽组织,阿托伐他汀组介于两者之间,轻度炎性细胞浸润,骨小梁略纤细,多数小梁骨陷窝内可见骨细胞,小梁间为血管和骨髓。MMP-2在激素组、阿托伐他汀组、对照组中的表达分别为0.15±0.04、0.10±0.09、0.09±0.03;MMP-9在3组中的表达分别为0.13±0.03,0.11±0.05和0.08±0.02;TIMP-1在3组中的表达分别为0.07±0

  19. ADAM12 redistributes and activates MMP-14, resulting in gelatin degradation, reduced apoptosis and increased tumor growth

    DEFF Research Database (Denmark)

    Albrechtsen, Reidar; Hansen, Dorte Stautz; Vikeså, Jonas;

    2013-01-01

    Matrix metalloproteinases (MMPs), in particular MMP-2, MMP-9 and MMP-14, play a key role in various aspects of cancer pathology. Likewise, ADAMs (a disintegrin and metalloproteinases), including ADAM12, are upregulated in malignant tumors and contribute to the pathology of cancers. Here, we show...

  20. Cadmium exposure inhibits MMP2 and MMP9 activities in the prostate and testis

    Energy Technology Data Exchange (ETDEWEB)

    Lacorte, Livia M.; Rinaldi, Jaqueline C.; Justulin, Luis A.; Delella, Flávia K. [Univ Estadual Paulista – UNESP, Institute of Biosciences, Department of Morphology, Extracellular Matrix Laboratory, Botucatu, SP (Brazil); Moroz, Andrei [Univ Estadual Paulista – UNESP, School of Pharmaceutical Sciences, Department of Bioprocess and Biotechnology, Cell Culture Laboratory, Araraquara, SP (Brazil); Felisbino, Sérgio L., E-mail: felisbin@ibb.unesp.br [Univ Estadual Paulista – UNESP, Institute of Biosciences, Department of Morphology, Extracellular Matrix Laboratory, Botucatu, SP (Brazil)

    2015-02-20

    Matrix metalloproteinases (MMPs) are zinc (Zn{sup 2+}) and calcium (Ca{sup 2+}) dependant endopeptidases, capable of degradation of numerous components of the extracellular matrix. Cadmium (Cd{sup 2+}) is a well known environmental contaminant which could impair the activity of MMPs. In this sense, this study was conducted to evaluate if Cd{sup 2+} intake inhibits these endopeptidases activities at the rat prostate and testicles and if it directly inhibits the activity of MMP2 and MMP9 at gelatinolytic assays when present in the incubation buffer. To investigate this hypothesis, Wistar rats (5 weeks old), were given tap water (untreated, n = 9), or 15 ppm CdCl{sub 2} diluted in drinking water, during 10 weeks (n = 9) and 20 weeks (n = 9). The animals were euthanized and their ventral prostate, dorsal prostate, and testicles were removed. These tissue samples were processed for protein extraction and subjected to gelatin zymography evaluation. Additionally, we performed an experiment of gelatin zymography in which 5 μM or 2 mM cadmium chloride (CdCl{sub 2}) was directly dissolved at the incubation buffer, using the prostatic tissue samples from untreated animals that exhibited the highest MMP2 and MMP9 activities in the previous experiment. We have found that CdCl{sub 2} intake in the drinking water led to the inhibition of 35% and 30% of MMP2 and MMP9 (p < 0.05) at the ventral prostate and testis, respectively, in Cd{sup 2+} treated animals when compared to controls. Moreover, the activities of the referred enzymes were 80% and 100% inhibited by 5 μM and 2 mM of CdCl{sub 2}, respectively, even in the presence of 10 mM of CaCl{sub 2} within the incubation buffer solution. These important findings demonstrate that environmental cadmium contamination may deregulate the natural balance in the extracellular matrix turnover, through MMPs downregulation, which could contribute to the toxic effects observed in prostatic and testicular tissue after its

  1. Regulation of E1AF on matrix metaloproteinase-9 in colorectal cancer%大肠癌E1AF基因的表达对MMP-9的调控作用

    Institute of Scientific and Technical Information of China (English)

    朱民; 朱军

    2005-01-01

    目的:探讨大肠癌患者血浆基质金属蛋白酶-9(MMP-9)和肿瘤组织中E1A因子(E1AF)表达的临床意义及两者之间的关系.方法:采用酶链免疫吸附法(ELISA)检测51例大肠癌患者血浆MMP-9浓度,采用RT-PCR方法检测肿瘤组织中E1AF的表达,进行定量和半定量分析.结果:Ⅲ、Ⅳ期大肠癌患者血浆MMP-9浓度和肿瘤组织中E1AF的表达水平高于Ⅰ、Ⅱ期患者(P<0.05),Ⅳ期大肠癌患者MMP-9的浓度高于Ⅲ期患者(P<0.05);血浆MMP-9浓度和肿瘤组织中E1AF的表达呈正相关(r=0.77,P<0.05).结论:患者血浆MMP-9和肿瘤组织中E1AF的高表达与大肠癌浸润转移有关,E1AF参与MMP-9的表达调控.

  2. 基质金属蛋白酶抑制剂2研究进展

    Institute of Scientific and Technical Information of China (English)

    纪明开; 林建银

    2008-01-01

    内源性金属蛋白酶组织抑制剂(tissue inhibitor of metallo- proteinase,TIMPs)为基质金属蛋白酶(matrix metalloprotease,MMP)特异性抑制剂,属半胱氨酸蛋白酶超家族,存在于果蝇、秀丽隐杆线虫、人等多种真核生物中,机体缺乏TIMP会引起胚胎发育异常。TIMPs N端功能区半胱氨酸残基与MMPs的锌离子活性中心结合,C端功能区与MMPs的其他部位结合,以1:1的比例非共价键结合形成MMP-TIMP复合体,

  3. The Expression and Significance of Matrix Metalloproteinase-2 and CD147 in Gastric Carcinoma%CD147及MMP-2在胃癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    黄悦; 赵建华; 樊克武

    2007-01-01

    目的 探讨CD147及基质金属蛋白酶-2(MMP-2)在胃癌中表达特点及其生物学行为关系.方法 应用免疫组织化学S-P法检测58例胃癌组织及20例正常胃黏膜组织中CD147和MMP-2的表达情况.结果 CD147及MMP-2在胃癌组中的阳性表达率分别为77.6%及82 8%,均高于正常胃黏膜组(P<0.01);两者的阳性表达均与胃癌的浸润深度、临床分期及淋巴结转移有关,与分化程度无明显关联;胃癌中CD147及MMP-2的表达具有相关性(P<0.01).结论 胃癌组织中CD147和MMP-2的高表达,可以作为胃癌生物学行为的客观参考指标.

  4. MMP-2、MMP-9在乳腺导管癌中表达及意义%Expression and significance of MMP-2, MMP-9 in breast carcinoma

    Institute of Scientific and Technical Information of China (English)

    苏书娟; 邢鲁奇; 陈登庭; 邓淼

    2011-01-01

    Objective To research the expression and relationship of matrix metalloproteinase-2 ( MMP-2), metrix metalloproteinase-9(MMP-9) and collagen Ⅳ in breast ductal carcinoma. Methods Immunohistochemistry double staining was used to detect the expression of MMP-2, MMP-9 and collagen Ⅳ in 60 cases of breast ductal carcinoma and analyze their correlation. The relations between MMP-2, MMP-9 and breast ductal carcinoma tumor size, lymph node metastasis, and C-erbB-2 were analyzed. Results The positive expression rate of MMP-2 ,MMP-9 in breast ductal carcinoma was 75% and 80% ,respectively,which were significantly higher than the rate of 10% in normal breast tissues. MMP-2 and MMP-9 were expressed more strongly in the cancer cells approaching the basement membrane or breaking through the basement membrane. The positive expressions of MMP-2 and MMp-9 in the tumor diameter > 2em, with lymph node metastasis, and C-erbB-2 positive group were 83.9% and 90. 3% ,87.0% and 91.3% ,and 80. 9% and 85. 1% ,which were significantly higher than 58.6% and 65.5% ,64. 9% and 67. 6% ,and 46. 2% and 53. 8% of the correponding control groups. Collagen Ⅳ expression and the expression of MMP-2, MMP-9 was negatively correlated. Conclusion The degradation of collagen Ⅳ related to MMP-2 and MMP-9 may play an important role in local invasive and distant metastasis of breast ductal carcinoma. The expressions of MMP-2 ,MMP-9 and collagen Ⅳ may have reference value in determining the invasive and metastatic potential of breast carcinoma and evaluating prognosis.%目的 研究乳腺导管癌中基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)的表达及与Ⅳ型胶原(collagenⅣ)的关系.方法 免疫组化双染法检测60例乳腺导管癌中MMP-2、MMP-9和Ⅳ型胶原的表达并分析其相关性及与肿瘤大小、淋巴结转移及人类表皮生长因子受体-2(C-erbB-2)的关系.结果 MMP-2、MMP-9在乳腺导管癌中

  5. The relationship of MMP-12 and the pulmonary disease%MMP-12与肺部疾病的关系

    Institute of Scientific and Technical Information of China (English)

    杨宁

    2010-01-01

    基质金属蛋白酶-12(MMP-12)是基质金属蛋白酶家族中的一员,是细胞外基质降解的主要介质,结构包括前肽域、催化域和血红素结合蛋白样域.MMP-12的作用主要是基因转录和抑制物的调节两个方面.MMP-12的过度表达导致病理的细胞外基质蛋白降解和过度的气道重塑,引起哮喘、肺气肿等许多呼吸系统疾病.%Matrix matalloprotenases-12(MMP-12) is a member of the matrix matalloprotenases family,and it is a main mediator to degradate the extracellular matrix. MMP-12 is composed of an propeptide domain, a catalytic domain and hemopexin-like domain. The function of MMP-12 include the genetic transcription and inhibitor regulation. Overexpress of MMP-12 causes pathological extracellular matrix protein breakdown and excessive airway remodeling,and MMP-12 is impotant in the pathogenesis of the acute and chronic lung injury,which has been implicated in a range of respiratory diseases, including asthma and emphysema.

  6. MT1-MMP expression level status dictates the in vitro action of lupeol on inflammatory biomarkers MMP-9 and COX-2 in medulloblastoma cells.

    Science.gov (United States)

    Annabi, Borhane; Vaillancourt-Jean, Eric; Béliveau, Richard

    2013-02-01

    Local inflammation-induced extracellular matrix structural changes are a prerequisite to neoplastic invasion by pediatric intracranial tumors. Accordingly, increased expression of matrix metalloproteinases MMP-2 and MMP-9, two inflammation-induced matrix metalloproteinases (MMPs), may further aid the transformed cells either to infiltrate adjacent tissues or to enter the peripheral circulation. In the context of neuroinflammation, MMP-9 has been linked to processes such as blood-brain barrier opening and invasion of neural tissue by blood-derived immune cells. Given its reported anti-inflammatory and anticancer properties, we investigated the in vitro pharmacological effects of lupeol, a diet-derived triterpenoid, on MMP-9 and cyclooxygenase (COX)-2 expressions in a pediatric medulloblastoma DAOY cell line model. Lupeol was unable to inhibit the increased MMP-9 and COX-2 expression in phorbol 12-myristate 13-acetate (PMA)-treated cells, but was rather found to synergize with PMA to induce both biomarkers' expression. A contribution of the membrane type-1 (MT1)-MMP was also revealed, since lupeol/PMA treatments triggered proMMP-2 activation, and that MT1-MMP gene silencing reversed the combined effects of lupeol/PMA on both MMP-9 and COX-2. The mRNA stabilizing factor HuR was also found increased in the combined lupeol/PMA treatment, suggesting stabilization processes of the MMP-9 and COX-2 transcripts. We postulate that lupeol's anti-inflammatory properties may exert better pharmacological action within low MT1-MMP expressing tumors. Furthermore, these evidences add up to the new pleiotropic molecular mechanisms of action of MT1-MMP, and prompt for evaluating the future in vitro pharmacological properties of lupeol under pro-inflammatory experimental set-up.

  7. Monitoring and Inhibiting MT1-MMP during Cancer Initiation and Progression

    Directory of Open Access Journals (Sweden)

    Sonia Pahwa

    2014-02-01

    Full Text Available Membrane-type 1 matrix metalloproteinase (MT1-MMP is a zinc-dependent type-I transmembrane metalloproteinase involved in pericellular proteolysis, migration and invasion. Numerous substrates and binding partners have been identified for MT1-MMP, and its role in collagenolysis appears crucial for tumor invasion. However, development of MT1-MMP inhibitors must consider the substantial functions of MT1-MMP in normal physiology and disease prevention. The present review examines the plethora of MT1-MMP activities, how these activities relate to cancer initiation and progression, and how they can be monitored in real time. Examination of MT1-MMP activities and cell surface behaviors can set the stage for the development of unique, selective MT1-MMP inhibitors.

  8. Polymorphisms of the MMP-9 gene and abdominal aortic aneurysm

    Science.gov (United States)

    Smallwood, Linda; Allcock, Richard; van Bockxmeer, Frank; Warrington, Nicole; Palmer, Lyle J; Iacopetta, Barry; Golledge, Jonathan; Norman, Paul E

    2008-01-01

    Background Increased matrix metalloproteinase-9 (MMP-9) activity has been implicated in the formation of abdominal aortic aneurysms (AAAs). The aim of the present study was to explore the association between potentially functional variants of the MMP-9 gene and AAA. Method The −1562C>T and −1811A>T variants of the MMP-9 gene were genotyped in 678 men with AAAs (>30mm in diameter) and 659 controls (aortic diameter 19−22mm) recruited from a population-based trial of screening for AAAs. The levels of MMP-9 were measured in a random subset of 300 cases and 84 controls. The association between genetic variants (including haplotypes) and AAA was assessed using multivariate logistic regression. Results There was no association between the MMP-9 −1562C>T (OR 0.70 95%CI 0.27, 1.82) or −1811A>T (OR 0.71, 95%CI 0.28, 1.85) genotypes, or the most common haplotype (OR 0.81 95%CI 0.62, 1.05), and AAA. The serum MMP-9 concentration (ng/mL) was higher in cases than controls and in minor allele carriers in cases and controls although the differences were not statistically significant. Conclusion The results suggest that a genetic tendency to have higher levels of circulating MMP-9 is not associated with AAAs. PMID:18763261

  9. Macrophage metalloelastase (MMP12) regulates adipose tissue expansion, insulin sensitivity, and expression of inducible nitric oxide synthase.

    Science.gov (United States)

    Lee, Jung-Ting; Pamir, Nathalie; Liu, Ning-Chun; Kirk, Elizabeth A; Averill, Michelle M; Becker, Lev; Larson, Ilona; Hagman, Derek K; Foster-Schubert, Karen E; van Yserloo, Brian; Bornfeldt, Karin E; LeBoeuf, Renee C; Kratz, Mario; Heinecke, Jay W

    2014-09-01

    Macrophage metalloelastase, a matrix metallopeptidase (MMP12) predominantly expressed by mature tissue macrophages, is implicated in pathological processes. However, physiological functions for MMP12 have not been described. Because mRNA levels for the enzyme increase markedly in adipose tissue of obese mice, we investigated the role of MMP12 in adipose tissue expansion and insulin resistance. In humans, MMP12 expression correlated positively and significantly with insulin resistance, TNF-α expression, and the number of CD14(+)CD206(+) macrophages in adipose tissue. MMP12 was the most abundant matrix metallopeptidase detected by proteomic analysis of conditioned medium of M2 macrophages and dendritic cells. In contrast, it was detected only at low levels in bone marrow derived macrophages and M1 macrophages. When mice received a high-fat diet, adipose tissue mass increased and CD11b(+)F4/80(+)CD11c(-) macrophages accumulated to a greater extent in MMP12-deficient (Mmp12(-/-)) mice than in wild-type mice (Mmp12(+/+)). Despite being markedly more obese, fat-fed Mmp12(-/-) mice were more insulin sensitive than fat-fed Mmp12(+/+) mice. Expression of inducible nitric oxide synthase (Nos2) by Mmp12(-/-) macrophages was significantly impaired both in vivo and in vitro, suggesting that MMP12 might mediate nitric oxide production during inflammation. We propose that MMP12 acts as a double-edged sword by promoting insulin resistance while combatting adipose tissue expansion.

  10. Expression of MMP-2 and MMP-9 in the rat trigeminal ganglion during the development of temporomandibular joint inflammation.

    Science.gov (United States)

    Nascimento, G C; Rizzi, E; Gerlach, R F; Leite-Panissi, C R A

    2013-11-18

    Orofacial pain is a prevalent symptom in modern society. Some musculoskeletal orofacial pain is caused by temporomandibular disorders (TMDs). This condition has a multi-factorial etiology, including emotional factors and alteration of the masticator muscle and temporomandibular joints (TMJs). TMJ inflammation is considered to be a cause of pain in patients with TMD. Extracellular proteolytic enzymes, specifically the matrix metalloproteinases (MMPs), have been shown to modulate inflammation and pain. The purpose of this investigation was to determine whether the expression and level of gelatinolytic activity of MMP-2 and MMP-9 in the trigeminal ganglion are altered during different stages of temporomandibular inflammation, as determined by gelatin zymography. This study also evaluated whether mechanical allodynia and orofacial hyperalgesia, induced by the injection of complete Freund's adjuvant into the TMJ capsule, were altered by an MMP inhibitor (doxycycline, DOX). TMJ inflammation was measured by plasma extravasation in the periarticular tissue (Evans blue test) and infiltration of polymorphonuclear neutrophils into the synovial fluid (myeloperoxidase enzyme quantification). MMP expression in the trigeminal ganglion was shown to vary during the phases of the inflammatory process. MMP-9 regulated the early phase and MMP-2 participated in the late phase of this process. Furthermore, increases in plasma extravasation in periarticular tissue and myeloperoxidase activity in the joint tissue, which occurred throughout the inflammation process, were diminished by treatment with DOX, a nonspecific MMP inhibitor. Additionally, the increases of mechanical allodynia and orofacial hyperalgesia were attenuated by the same treatment.

  11. Expression of MMP-2 and MMP-9 in the rat trigeminal ganglion during the development of temporomandibular joint inflammation

    Directory of Open Access Journals (Sweden)

    G.C. Nascimento

    2013-11-01

    Full Text Available Orofacial pain is a prevalent symptom in modern society. Some musculoskeletal orofacial pain is caused by temporomandibular disorders (TMDs. This condition has a multi-factorial etiology, including emotional factors and alteration of the masticator muscle and temporomandibular joints (TMJs. TMJ inflammation is considered to be a cause of pain in patients with TMD. Extracellular proteolytic enzymes, specifically the matrix metalloproteinases (MMPs, have been shown to modulate inflammation and pain. The purpose of this investigation was to determine whether the expression and level of gelatinolytic activity of MMP-2 and MMP-9 in the trigeminal ganglion are altered during different stages of temporomandibular inflammation, as determined by gelatin zymography. This study also evaluated whether mechanical allodynia and orofacial hyperalgesia, induced by the injection of complete Freund's adjuvant into the TMJ capsule, were altered by an MMP inhibitor (doxycycline, DOX. TMJ inflammation was measured by plasma extravasation in the periarticular tissue (Evans blue test and infiltration of polymorphonuclear neutrophils into the synovial fluid (myeloperoxidase enzyme quantification. MMP expression in the trigeminal ganglion was shown to vary during the phases of the inflammatory process. MMP-9 regulated the early phase and MMP-2 participated in the late phase of this process. Furthermore, increases in plasma extravasation in periarticular tissue and myeloperoxidase activity in the joint tissue, which occurred throughout the inflammation process, were diminished by treatment with DOX, a nonspecific MMP inhibitor. Additionally, the increases of mechanical allodynia and orofacial hyperalgesia were attenuated by the same treatment.

  12. MMP-26及HPSE在子宫颈癌中的表达及临床意义%Expressions of matrix metalloproteinase-26 and heparanase in cervical cancer and the clinical significance

    Institute of Scientific and Technical Information of China (English)

    齐彩霞; 杨秀兰; 穆雅琴; 席国萍; 朱壮彦

    2015-01-01

    目的 探讨MMP-26及HPSE在正常子宫颈、原位癌、浸润癌中的表达及临床意义.方法 采用免疫组织化学法检测MMP-26及HPSE在15例正常子宫颈(正常宫颈组)、21例宫颈原位癌(宫颈原位癌组)、36例宫颈早期浸润癌(宫颈早期浸润癌组)患者中的表达情况.结果 原位癌及早期浸润癌组患者中MMP-26及HPSE的表达明显高于正常宫颈组,差异有统计学意义(P<0.01);二者早期浸润癌组中的表达明显高于原位癌组,差异有统计学意义(P<0.05).结论 MMP-26及HPSE在宫颈癌的浸润与转移过程发挥着重要作用,可以作为判断患者预后的指标.

  13. Involvement of TSP1 and MMP9/NGAL in Angiogenesis during Orthodontic Periodontal Remodeling

    Directory of Open Access Journals (Sweden)

    Petra Surlin

    2014-01-01

    Full Text Available In the present study the aim was to measure the levels of Thrombospondin-1 (TSP1 and Lipocalin-2/matrix metalloproteinase 9 (MMP9/NGAL complex in gingival crevicular fluid (GCF at different time points of orthodontic treatment, to determine the relationship between these values and those of total-matrix metalloproteinase 9 (MMP9 and theirs implication in angiogenesis balance, in the situation of a good control of the bacterial plaque, emphasizing the role of TSP1 and MMP9/NGAL complex. GCF samples were collected from 16 young orthodontic patients requiring upper canine distalization (test tooth with first premolar extraction. The contralateral canine (control tooth was free from orthodontic force. For the orthodontic appliance, brackets Roth 0.018 inch with 0.012 inch NiTi archwire and a laceback were used. TSP1, MMP9/NGAL, and MMP9 increased from 1 hour before activation of orthodontic appliance to a maximum at 8 hours for MMP9 and 72 hours for MMP9/NGAL and TSP1. The results show a change in time of TSP1, MMP9/NGAL, and MMP9 levels in GCF of patients with this method of orthodontic treatment. The powerful correlation of MMP9/NGAL with TSP1 suggests their stronger involvement in angiogenesis processes in PDL during orthodontic periodontal remodeling, in the situation of a healthy periodontium and a good control of the bacterial plaque.

  14. SPDEF 调控小鼠成釉细胞MMP-20基因表达的分子机制研究%Study of Transcription Factor SPDEF on Regulating the Expression of Matrix Metalloproteinase-20

    Institute of Scientific and Technical Information of China (English)

    纪虹利; 杜森; 董超; 张世龙; 靖旭; 高玉光

    2013-01-01

    Objective To analyse the influence of the expression of MMP-20 gene by cloning the transcription factor SPDEF gene and constructing eukaryotic expression vector in order to further study the role of transcription factor SPDEF in the formation of the en -amel.Methods The SPDEF gene was acquired from ALC by RT-PCR,Then inserted into pcDNA3.1/myc-HisA vector.The recombinant plasmid was transiently transfected into ALC .Dual-Luciferase Reporter Assay System was performed to detect the transcriptional activity of MMP-20 promoter region.Results ①SPDEF was amplification by RT-PCR and matched the expected fragment about 978bp.The sequencing result was completely matched with the known sequence in GeneBank accession NO .NM:013891.4.②The recombinant plasmid of pcDNA3. 1/myc-SPDEF was transfect into ALC.Then detect the increase of the transcriptional activity of MMP-20.Conclusion The eukaryotic ex-pression vector carried SPDEF is constructed successfully .Preliminary analysis has shown that SPDEF may be combined with the characteris-tic sequence of MMP-20 which up-regulate the expression of MMP-20.%  目的通过克隆转录因子SPDEF以及其真核表达载体的构建,分析该基因对成釉细胞MMP-20基因表达的影响,为进一步研究转录因子SPDEF在牙釉质形成中发挥的作用奠定基础.方法利用RT-PCR技术从小鼠成釉细胞中获得SPDEF基因,测序正确后克隆至pcDNA3.1/myc-HisA载体中,将重组质粒瞬时转染成釉细胞,并用双荧光素酶报告基因系统检测技术来检测MMP-20启动子区域的转录活性.结果①SPDEF基因经过RT-PCR扩增得到的产物与预期片段978bp相符,将所获得的目的基因与GenBank 提供的已知序列(NM:013891.4)完全一致.②重组质粒pcDNA3.1/myc-HisA-SPDEF转染成釉细胞后,用荧光素酶分析系统检测显示MMP-20启动子转录活性升高.结论成功构建了SPDEF真核表达载体,初步研究证明SPDEF可能与MMP-20特征性序列结合从而上调MMP

  15. Association of MMP-9 gene polymorphisms with nephrolithiasis patients.

    Science.gov (United States)

    Mehde, Atheer Awad; Mehdi, Wesen Adel; Yusof, Faridah; Raus, Raha Ahmed; Zainal Abidin, Zaima Azira; Ghazali, Hamid; Abd Rahman, Azlina

    2017-02-15

    Nephrolithiasis is one of the causes which lead to chronic kidney disease (CKD). Matrix metalloproteinases (MMPs) are endopeptidases degrading extracellular matrix which correlate with the pathogenesis of atherosclerosis. The current study was designed to analyze the association of (R279Q, C1562T) polymorphism of MMP-9 with nephrolithiasis patients. Genotyping of MMP-9/R279Q and of MMP-9/C1562T polymorphism were carried out by PCR-based restriction digestion method. Serum level of MMP-9, oxidative stress marker, MDA, and uric acid were measured in patients and control. Allele frequencies of the MMP-9/C1562T polymorphism for C and T allele were 71.25% and 28.75% in patients, 87.08% and 12.92% in control respectively. The homozygote TT was more frequent in the nephrolithiasis patients group, while T allele frequency was significantly higher in the nephrolithiasis patients group than in the control group. The patients with CT and TT genotype showed a significant increase in serum MMP-9, Total Oxidant Status (TOS), Oxidative Stress Index (OSI), Malondialdehyde (MDA), and uric acid when compared to CC genotype in patients with nephrolithiasis. The R279Q polymorphism site with regard to the relationship with nephrolithiasis was not significant. The result indicates that patients with TT genotype had an increased risk of stones. Also, the results demonstrate that TT allele of the C1562T polymorphism in the MMP-9gene is related with an increase of oxidative stress in nephrolithiasis patients and may possibly impose a risk for cardiovascular diseases in patients with TT genotype of MMP-9. © 2017 Wiley Periodicals, Inc.

  16. Accelerated neointima formation after vascular injury in mice with stromelysin-3 (MMP-11) gene inactivation.

    Science.gov (United States)

    Lijnen, H R; Van Hoef, B; Vanlinthout, I; Verstreken, M; Rio, M C; Collen, D

    1999-12-01

    The hypothesis that stromelysin-3 (MMP-11), a unique member of the matrix metalloproteinase (MMP) family, plays a role in neointima formation was tested with the use of a vascular injury model in wild-type (MMP-11(+/+)) and MMP-11-deficient (MMP-11(-/-)) mice. Neointima formation 2 to 3 weeks after electric injury of the femoral artery was significantly enhanced in MMP-11(-/-) as compared with MMP-11(+/+) mice, in both mice of a pure 129SV genetic background (0.014 versus 0.0010 mm(2) at 2 weeks, P<0.001) and those of a 50/50 mixed 129SV/BL6 background (0.030 versus 0.013 mm(2) at 3 weeks, P<0.05). The medial areas were comparable, resulting in intima/media ratios that were significantly increased in MMP-11(-/-) as compared with MMP-11(+/+) arteries, in mice of both the 129SV (1. 0 versus 0.18, P<0.001) and mixed (1.5 versus 0.70, P<0.05) backgrounds. Nuclear cell counts in cross-sectional areas of the intima of the injured region were higher in arteries from MMP-11(-/-) mice than in those from MMP-11(+/+) mice (210 versus 48, P<0.001, in pure 129SV mice and 290 versus 150, P<0.01, in mice of the mixed genetic background). Immunocytochemical analysis revealed that alpha-actin-positive and CD45-positive cells were more abundant in intimal sections of MMP-11(-/-) mice. Degradation of the internal elastic lamina was more extensive in arteries of MMP-11(-/-) mice than in those of MMP-11(+/+) mice (39% versus 6.8% at 3 weeks, P<0. 005). The mechanisms by which MMP-11 could impair elastin degradation and cellular migration in this model remain, however, unknown.

  17. 下肢曲张静脉壁MMP-2、MMP-9和胶原含量的临床研究%Clinical study of MMP-2, MMP-9 and collagen in lower limb varicose veins

    Institute of Scientific and Technical Information of China (English)

    邵拥军; 朱化刚; 周静

    2012-01-01

    目的研究曲张大隐静脉管壁基质金属蛋白酶(MMP)-2、MMP-9的表达及其与临床症状和体征分期的关系,并检测曲张静脉壁总的胶原含量,探讨MMP-2、MMP-9和胶原在曲张静脉重塑时的作用.方法采用免疫组化SP法检测曲张静脉壁和正常大隐静脉壁MMP-2和MMP-9的表达,采用Masson染色法检测曲张静脉壁和正常对照组总的胶原含量,采用χ2检验和t检验进行统计学分析处理.结果 MMP-2和MMP-9在正常静脉壁和曲张静脉壁均有表达,但曲张静脉壁MMP-2和MMP-9的阳性率明显升高(P<0.05);C4-C6期MMP-2和MMP-9的阳性率明显高于C1-C3期(P<0.05);曲张静脉壁总的胶原含量显著高于正常对照组(P<0.05).结论 MMP-2、MMP-9的异常表达导致了静脉壁总的胶原含量增加,参与了曲张静脉的重塑.%Objective To investigate the expression of matrix metalloproteinase -2 and -9 in varicose veins, and their relationships with clinical stages , and to discuss the changes of total collagen in varicose veins, and their significance. Methods The expression of MMP-2 and MMP-9 was examined by immunohistochemistry SP methods in varicose veins (VV) and normal veins (NVV), the total collagen was detected using Masson staining in varicose veins and normal veins, and chi-square test and T- test was used for statistical analysis. Results MMP-2 and -9 were both expressed in NVV and VV, and the positive expression rate in VV was significantly higher than that in NVV (P< 0.05). The positive expression of MMP-2 and -9 was significantly higher in C4-C6 stages than in C1-C3 stages (P <0.05), and the collagen was significantly increased in varicose veins (P <0.05). Conclusion The abnormal expression of MMP-2 and -9 leads to collagen increasing in varicose veins, which may participate in vascular remodeling in varicose veins.

  18. Active G protein-coupled receptors (GPCR), matrix metalloproteinases 2/9 (MMP2/9), heparin-binding epidermal growth factor (hbEGF), epidermal growth factor receptor (EGFR), erbB2, and insulin-like growth factor 1 receptor (IGF-1R) are necessary for trenbolone acetate-induced alterations in protein turnover rate of fused bovine satellite cell cultures.

    Science.gov (United States)

    Thornton, K J; Kamanga-Sollo, E; White, M E; Dayton, W R

    2016-06-01

    Trenbolone acetate (TBA), a testosterone analog, increases protein synthesis and decreases protein degradation in fused bovine satellite cell (BSC) cultures. However, the mechanism through which TBA alters these processes remains unknown. Recent studies indicate that androgens improve rate and extent of muscle growth through a nongenomic mechanism involving G protein-coupled receptors (GPCR), matrix metalloproteinases (MMP), heparin-binding epidermal growth factor (hbEGF), the epidermal growth factor receptor (EGFR), erbB2, and the insulin-like growth factor-1 receptor (IGF-1R). We hypothesized that TBA activates GPCR, resulting in activation of MMP2/9 that releases hbEGF, which activates the EGFR and/or erbB2. To determine whether the proposed nongenomic pathway is involved in TBA-mediated alterations in protein turnover, fused BSC cultures were treated with TBA in the presence or absence of inhibitors for GPCR, MMP2/9, hbEGF, EGFR, erbB2, or IGF-1R, and resultant protein synthesis and degradation rates were analyzed. Assays were replicated at least 9 times for each inhibitor experiment utilizing BSC cultures obtained from at least 3 different steers that had no previous exposure to steroid compounds. As expected, fused BSC cultures treated with 10 n TBA exhibited increased ( BSC cultures with 10 n TBA in the presence of inhibitors for GPCR, MMP2/9, hbEGF, EGFR, erbB2, or IGF-1R suppressed ( 0.05) effect on TBA-mediated decreases in protein degradation. However, inhibition of both EGFR and erbB2 in the presence of 10 n TBA resulted in decreased ( BSC cultures treated with 10 n TBA exhibit increased ( BSC cultures.

  19. Grafting MAP peptide to dental polymer inhibits MMP-8 activity.

    Science.gov (United States)

    Dixit, Namrata; Settle, Jenifer K; Ye, Qiang; Berrie, Cindy L; Spencer, Paulette; Laurence, Jennifer S

    2015-02-01

    Matrix metalloproteinases (MMPs) are a class of zinc and calcium-dependent endopeptidases responsible for degrading extracellular matrix (ECM) components. Their activity is critical for both normal biological function and pathological processes (Dejonckheere et al., Cytokine Growth Factor Rev 2011;22:73-81). In dental restorations, the release and subsequent acid activation of MMPs contributes to premature failure. In particular, MMP-8 accelerates degradation by cleaving the collagen matrix within the dentin substrate in incompletely infiltrated aged bonded dentin (Buzalaf et al., Adv Dent Res 2012;24:72-76), hastening the need for replacement of restorations. Therefore, development of a dental adhesive that better resists MMP-8 activity is of significant interest. We hypothesize that modification of the polymer surface with an inhibitor would disable MMP-8 activity. Here, we identify the metal abstraction peptide (MAP) as an inhibitor of MMP-8 and demonstrate that tethering MAP to methacrylate polymers effectively inhibits catalysis. Our findings indicate complete inhibition of MMP-8 is achievable using a grafting approach. This strategy has potential to improve longevity of dental adhesives and other polymers and enable rational design of a new generation of biocompatible materials.

  20. Effect of human osteopontin on proliferation, transmigration and expression of MMP-2 and MMP-9 in osteosarcoma cells

    Institute of Scientific and Technical Information of China (English)

    刘思金; 胡国法; 刘亚军; 刘思国; 高虹; 张传生; 魏影允; 薛延; 劳为德

    2004-01-01

    Background To explore the effect of human osteopontin (hOPN) on the proliferation, transmigration and expression of matrix metallproteinase-2 (MMP-2) and matrix metallproteinase-9 (MMP-9) in osteosarcoma (OS) cells in vitro. Methods The prokaryotic-expression vector of hOPN was produced, hOPN was then subcloned into E. coli BL21 (DE3) cells and purified with ProBondTM Columns. The proliferation, cell cycle and the expression of cyclin A in OS cells were investigated by using MTT assay, flow cytometry and Western blot respectively. The transmigration of OS cells was checked by using transwell cell culture chamber. The micro-pore-filter-membrane system was used to study the chemiotaxis of hOPN to OS cells. The levels of total protein were examined according to Coomassie Brilliant Blue manuals. The expression of MMP-2 and MMP-9 were evaluated by detecting the volume of degradation of gelatin on SDS-PAGE gel.Results The prokaryotic-expression vector of hOPN and purified hOPN protein were achieved hOPN promoted OS cells proliferation in a dose-dependent manner, and stimulated cyclin A expression in OS cells to accelerate cell division cycle, hOPN facilitated the trans-membrane migration of OS cells. hOPN also enhanced the secretion of MMP-2 and MMP-9 in OS cells. Conclusion hOPN could stimulate cyclin A expression in OS cells, hOPN has chemiotaxis to OS cells and increases their transmigration, hOPN enhances the secretion of MMP-2 and MMP-9 in OS cells.

  1. Changes in expression of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in pathological scar tissue of rabbit ears after scar-skin replantation%兔耳病理性瘢痕皮回植术后组织中基质金属蛋白酶-1及金属蛋白酶组织抑制剂-1表达的变化

    Institute of Scientific and Technical Information of China (English)

    赵雪莲; 苏晓光; 张锤; 张春莉; 张卓男; 郑宝恒

    2014-01-01

    目的 通过建立兔耳模型观察基质金属蛋白酶-1 (matrix metalloproteinase-1,MMP-1)及金属蛋白酶组织抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)在病理性瘢痕皮回植术后组织中表达的变化,探讨瘢痕皮回植治疗病理性瘢痕的机制.方法 建立兔耳病理性瘢痕模型,共分为3组:正常皮肤组(对照组,A组)、病理性瘢痕组(B组)及瘢痕皮回植组(C组).切取标本行HE染色和Masson特殊组织化学染色及免疫组织化学染色,观察各组标本MMP-1、TIMP-1的表达情况.结果 病理性瘢痕经瘢痕皮回植术后,MMP-1及TIMP-1均较A组明显升高(P<0.01),MMP-1的表达较TIMP-1明显增强(P<0.01).结论 瘢痕皮回植术治疗瘢痕的机制与瘢痕组织内MMP-1和TIMP-1相互作用的失衡有关.%Objective To observe the changes of MMP-1 and TIMP-1 expression in the pathological scar tissue after scar-skin replantation and to explore the mechanism of treating pathological scars with scar-skin replantation through a rabbit ear model.Methods Rabbit ears were used to establish the hypertrophic scar animal model in this study.Specimens were taken for three times:normal skin,hypertrophic scar and scar-skin replantion separately.We then performed HE staining,Masson staining and immunohistochemical staining to observe the expression of MMP-1 and TIMP-1 in these three groups of specimens.Results The expression of both MMP-1 and TIMP-1 significantly increased in the hypertrophic scar tissue after scar-skin replantation compared with the control group (P<0.01).The expression of MMP-1 increased more significantly than that of TIMP-1 (P<0.01).Conclusions The mechanism of scar-skin replantation's effect in the treatment for hypertrophic scar is relevant to the imbalance in the interaction between MMP-1 and TIMP-1 in the scar tissue.

  2. Functions of KLK4 and MMP-20 in dental enamel formation

    Science.gov (United States)

    Lu, Yuhe; Papagerakis, Petros; Yamakoshi, Yasuo; Hu, Jan C-C.; Bartlett, John D.; Simmer, James P.

    2009-01-01

    Two proteases are secreted into the enamel matrix of developing teeth. The early protease is enamelysin (MMP-20). The late protease is kallikrein 4 (KLK4). Mutations in MMP20 and KLK4 both cause autosomal recessive amelogenesis imperfecta, a condition featuring soft, porous enamel containing residual protein. MMP-20 is secreted along with enamel proteins by secretory stage ameloblasts. Enamel protein cleavage products accumulate in the space between the crystal ribbons, helping to support them. MMP-20 steadily cleaves accumulated enamel proteins, so their concentration decreases with depth. Kallikrein 4 is secreted by transition and maturation stage ameloblasts. KLK4 aggressively degrades the retained organic matrix following the termination of enamel protein secretion. The principle functions of MMP-20 and KLK4 in dental enamel formation are to facilitate the orderly replacement of organic matrix with mineral, generating an enamel layer that is harder, less porous, and unstained by retained enamel proteins. PMID:18627287

  3. 类风湿关节炎和骨关节炎滑膜滑液中MMP-2和MMP-9的比较%Comparison of the MMP-2 and MMP-9 in the Knee Joint Synovial Fluid and Synovial Membrane in Patients with active Rheumatoid Arthritis and Osteoarthritis

    Institute of Scientific and Technical Information of China (English)

    刘荣清; 林佳静; 孙伯坚; 李海波; 宋婷阁; 韩梅

    2013-01-01

    目的 检测基质金属蛋白酶-2(matrix metalloproteinases-2,MMP-2)和基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)在类风湿关节炎活动期(rheumatoid arthritis,RA)和骨关节炎(osteoarthritis,OA)活动期膝关节滑液及滑膜中的含量,并探讨两者在RA和OA病变中的作用.方法 收集宁夏医科大学总医院风湿免疫科和骨科确诊的14例RA活动期患者和24例OA活动期患者的膝关节滑液和滑膜组织,用酶联免疫吸附实验(ELISA)检测滑液中MMP-2和MMP-9的含量,用免疫组化法检测滑膜组织中MMP-9的相对表达量,组间比较采用秩和检验.结果 RA活动期患者膝关节滑液中的MMP-2和MMP-9浓度明显高于骨关节炎活动期患者(P<0.01);RA活动期患者膝关节滑膜中MMP-9的相对表达量明显高于OA活动期患者(P<0.01).结论 MMP-2和MMP-9在RA活动期患者滑液中高表达,MMP-9在RA活动期患者滑膜中高表达,可能参与RA发生发展过程中滑膜炎、骨质破坏.%Objective To compare matrix metalloproteinases - 2 ( MMP - 2) and matrix metalloproteinases - 9 ( MMP - 9) in knee joint synovial fluid and synovial membrane in patients with active rheumatoid arthritis ( RA) and active osteoarthritis ( OA) , and to explore the effects of MMP -2 and MMP -9 on lesion of RA and OA. Methods Knee joint synovial fluid and synovial membrane were collected from 14 and 24 patients with final diagnosis of RA and OA, and underwent surgery at the General Hospital of Ningxia Medical University. MMP - 2 and MMP - 9 in the synovial fluid were measured by enzyme linked immunosorbent assay (ELISA). The distributions of MMP - 9 in the synovial membrane were analyzed using immunohistochemistry. Student's t - test was used for intergroup comparison. Results The level of MMP - 2 and MMP - 9 in synovial fluid of active RA was significantly higher than that of active OA(P <0. 01) ; the relative expression of MMP -9 in synovial membrane was stronger in active RA

  4. Early MT-1 MMP expression following elastase exposure is associated with increased cleaved MMP-2 activity in experimental rodent aortic aneurysms.

    Science.gov (United States)

    Sinha, Indranil; Hannawa, Kevin K; Eliason, Jonathan L; Ailawadi, Gorav; Deogracias, Michael P; Bethi, Siddharth; Ford, John W; Roelofs, Karen J; Grigoryants, Vladimir; Henke, Peter K; Stanley, James C; Upchurch, Gilbert R

    2004-08-01

    The objective of this study was to determine the significance of membrane type 1 matrix metalloproteinase (MT1-MMP) activation of MMP-2 in experimental abdominal aortic aneurysms. Rat aortas were perfused with either saline as a control or elastase, and harvested on 2, 4, or 7 days after perfusion (n = 5 per treatment group/day). Aortic MT1-MMP and MMP-2 expression and protein were determined by real time polymerase chain reaction and Western blotting, respectively. Aortic explants were used to measure MMP-2 activity by zymography. Rat aortic smooth muscle cells in vitro were exposed to increasing doses of elastase and analyzed for MT-1 MMP expression. Aneurysms formed in 80% of the elastase-perfused aortas at 7 days, whereas none formed in the saline-perfused aortas. Significantly increased MT1-MMP expression was observed only on day 4, when levels were 6.5-fold higher in elastase-perfused aortas compared with saline-perfused aortas (P < .01). By day 7, MT1-MMP protein was present only in the elastase-perfused aortas (P = .02). By immunohistochemistry, MT1-MMP was detectable only in the elastase-perfused group at day 7. Cleaved MMP-2 activity (P = .045) was increased in elastase-perfused aortas compared with saline perfused aortas at day 7. In rat aortic smooth muscle cells, MT-1 MMP expression increased in response to elastase (P = .02). The rodent aortic aneurysm model exhibits upregulation of MT1-MMP expression and protein with subsequent increased conversion of MMP-2 from the latent to the cleaved form. Copyright 2004 Elsevier Inc.

  5. 子宫内膜腺癌中MMP-2、MMP-9表达的免疫组化研究%Metalloproteinase-9(MMP-9)in Endometrial Adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    仲肖静; 颜丽丽

    2014-01-01

    目的:探讨基质金属蛋白酶-2、-9(Matrix metalloproteinases,MMP)在子宫内膜腺癌组织中的表达及其与临床病理关系。方法:应用免疫组化SP法检测MMP-2、MMP-9在不同子宫内膜组织中的表达情况。结果:MMP-2在正常子宫内膜、子宫内膜增生症和子宫内膜腺癌组织中的阳性表达率分别为15.0%、26.7%、88.0%, MMP-9的阳性表达率分别为25.0%、33.3%、95.0%,MMP-2和MMP-9的表达在子宫内膜腺癌中均明显高于正常子宫内膜、子宫内膜增生症,比较差异均有统计学意义(P<0.05)。MMP-2、MMP-9的表达强度与子宫内膜腺癌的手术-病理分期、组织学分级、肌层浸润和淋巴结转移相关,差异均有统计学意义(P<0.05)。结论:MMP-2、MMP-9在子宫内膜腺癌的发生、发展过程中起促进作用,且两者表达与子宫内膜腺癌的浸润深度和转移程度有关,为早期诊断子宫内膜腺癌以及子宫内膜腺癌的预后判断、靶向治疗提供了可靠的依据。%Objective:To investigate the expression of MMP-2 and MMP-9 in endometrial adenocarcinoma tissue and its relation to clinical pathological characteristics.Method:Expression of MMP-2 and MMP-9 was examined by immunohistochemistry(SP method)in three different kinds of endometrium.Result:The results showed that the positive rates of MMP-2 in normal endometria,endometrial hyperplasias and endometrial adenocarcinomas were 15.0%,26.7%and 88.0%respectively,the positive rates of MMP-9 were 25.0%,33.3%and 95.0%respectively.Expression of MMP-2 and MMP-9 were significantly higher in endometrial adenocarcinoma than those in hyperplastic and normal endometria, the differences were statistically significant(P<0.05).In endometrial adenocarcinoma tissue,the positive expression of MMP-2 and MMP-9 was correlated with clinical pathological characteristics,histodifferentiation,myometrial invasion and lymphatic metastasis,the differences were statistically

  6. Concomitant lack of MMP9 and uPA disturbs physiological tissue remodeling

    DEFF Research Database (Denmark)

    Lund, Ida K; Nielsen, Boye S; Almholt, Kasper

    2011-01-01

    Urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP9, gelatinase B) have separately been recognized to play important roles in various tissue remodeling processes. In this study, we demonstrate that deficiency for MMP9 in combination with ablation of either uPA- or tissue...

  7. Tumor MMP-1 Activates Endothelial PAR1 to Facilitate Vascular Intravasation and Metastatic Dissemination

    DEFF Research Database (Denmark)

    Juncker-Jensen, Anna; Deryugina, Elena I; Rimann, Ivo

    2013-01-01

    non-tumor-cell/non-matrix target, activation of which by carcinoma-produced MMP-1 regulates endothelial permeability and transendothelial migration. The inhibitory effects of specific PAR1 antagonists in live animals have also indicated that the mechanisms of MMP-1-dependent vascular permeability...

  8. MMP-13 regulates growth of wound granulation tissue and modulates gene expression signatures involved in inflammation, proteolysis, and cell viability.

    Directory of Open Access Journals (Sweden)

    Mervi Toriseva

    Full Text Available Proteinases play a pivotal role in wound healing by regulating cell-matrix interactions and availability of bioactive molecules. The role of matrix metalloproteinase-13 (MMP-13 in granulation tissue growth was studied in subcutaneously implanted viscose cellulose sponge in MMP-13 knockout (Mmp13(-/- and wild type (WT mice. The tissue samples were harvested at time points day 7, 14 and 21 and subjected to histological analysis and gene expression profiling. Granulation tissue growth was significantly reduced (42% at day 21 in Mmp13(-/- mice. Granulation tissue in Mmp13(-/- mice showed delayed organization of myofibroblasts, increased microvascular density at day 14, and virtual absence of large vessels at day 21. Gene expression profiling identified differentially expressed genes in Mmp13(-/- mouse granulation tissue involved in biological functions including inflammatory response, angiogenesis, cellular movement, cellular growth and proliferation and proteolysis. Among genes linked to angiogenesis, Adamts4 and Npy were significantly upregulated in early granulation tissue in Mmp13(-/- mice, and a set of genes involved in leukocyte motility including Il6 were systematically downregulated at day 14. The expression of Pdgfd was downregulated in Mmp13(-/- granulation tissue in all time points. The expression of matrix metalloproteinases Mmp2, Mmp3, Mmp9 was also significantly downregulated in granulation tissue of Mmp13(-/- mice compared to WT mice. Mmp13(-/- mouse skin fibroblasts displayed altered cell morphology and impaired ability to contract collagen gel and decreased production of MMP-2. These results provide evidence for an important role for MMP-13 in wound healing by coordinating cellular activities important in the growth and maturation of granulation tissue, including myofibroblast function, inflammation, angiogenesis, and proteolysis.

  9. 胎盘组织中基质金属蛋白酶-9的表达与早产的关系%Relation of expressions of matrix metalloproteinase (MMP)-9 in human placenta and to preterm labor

    Institute of Scientific and Technical Information of China (English)

    段永红; 伍招娣

    2006-01-01

    目的探讨胎盘组织中MMp-9的表达与早产分娩发动的关系.方法采用免疫组化法(SABC技术)检测早产临产(PL)组(n=20)、早产未临产(PNL)组(n=20)、足月临产(TL)组(n=20)和足月未临产(TNL)组(n=20)4组胎盘组织中MMP-9的表达,根据Bresalier'半计量公式计算其平均染色强度.结果MMP-9在PL组、PNL组、TL组、TNL组胎盘组织中的平均染色强度分别是17.54±3.32、10.39±2.70、18.63±2.91和10.05±2.69,其中PL组与PNL组、TL组与TNL组之间的差异均有统计学意义(P<0.01);PL组与TL组、PNL组与TNL组之间的差异均无统计学意义(P>0.05).结论MMP-9的变化规律提示胎盘组织中MMP-9与分娩发动(早产或足月产)有关;早产与足月产具有类似的发动机制.

  10. 慢性阻塞性肺疾病患者肺血管重建与MMP-9及TIMP-1表达的关系%Relationships between the expressions of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 and lung revascularization in patients with chronic obstructive pulmonary disease

    Institute of Scientific and Technical Information of China (English)

    丛金鹏; 董蕾; 张馨文; 于文成

    2013-01-01

    目的:探讨慢性阻塞性肺疾病患者肺血管中基质金属蛋白酶-9(MMP-9)及其组织金属蛋白酶抑制剂-1(TIMP-1)的表达与肺血管重建的关系及临床意义。方法取60例因肺鳞癌行肺叶切除的癌旁组织标本,其中慢性阻塞性肺疾病组30例和对照组30例,图像分析系统测定肺动脉管壁面积/管总面积(WA%)、管壁厚度/血管外径(WT%)。免疫组化法测定肺血管平滑肌细胞MMP-9、TIMP及平滑肌细胞增殖细胞核抗原(PCNA)表达,并分析其与血管重建相关性。结果(1)慢性阻塞性肺疾病组肺血管炎症程度、WA%、WT%显著高于对照组(P<0.01)。(2)肺血管平滑肌细胞PCNA、MMP-9、TIMP-1的表达水平与对照组相比明显升高(P<0.01)。(3)相关分析:慢性阻塞性肺疾病组MMP-9、TIMP-1表达水平与血管炎症程度、WA%、WT%及PCNA呈显著正相关(P<0.01),多重线性回归分析显示MMP-9、TIMP-1表达水平的上调是影响肺血管重建(WA%、WT%)的主要因素(P<0.01)。结论 MMP-9、TIMP-1表达增强引起细胞外基质异常降解,诱导平滑肌增殖,促进炎症细胞迁移,在肺血管重塑过程中起到了重要作用。%Objective To explore the correlation between the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) and lung revascularization in patients with chronic obstructive pulmonary disease(COPD). Methods Lung tissues from patients with COPD(COPD group, n=30) and those without COPD(non-COPD group, n=30) were obtained from surgically resected specimens.The ratio of the thickness of the wall to the external diameter of the pulmonary arterioles(WT%) and the ratio of the area of the wall to that of the pulmonary arterioles(WA%) were analyzed by computer-based image analysis system. Immunohistochemical technique was applied to investigate the expressions of proliferating cell nuclear

  11. JNK/c-Jun signaling pathway mediates the fluoride-induced down-regulation of MMP-20 in vitro

    Science.gov (United States)

    Zhang, Yan; Li, Wu; Chi, Hae Sun; Chen, James; DenBesten, Pamela K.

    2008-01-01

    Delayed removal of amelogenins, which are initially hydrolyzed by matrix metalloproteinase MMP-20, is a characteristic of enamel fluorosis. In this study, we investigated the regulation of MMP-20 and possible effects of fluoride on MMP-20 expression in human ameloblast lineage cells. Protein expression and signaling pathways of human ameloblast lineage cells, exposed to 10 μM fluoride, were compared to control cells without fluoride exposure. The role of activator protein-1 in MMP-20 regulation was analyzed by DNA-protein affinity precipitation and luciferase reporter gene assays. MMP-20 protein levels in human ameloblast lineage cells decreased in the presence of fluoride, while amelogenin and TIMP-2 were not altered. Fluoride also decreased the transcription of a luciferase reporter gene driven by the MMP-20 promoter. Down-regulation of MMP-20 by fluoride was related to suppression of JNK/c-Jun phosphorylation. In contrast, the JNK activator elevated the expression of MMP-20. Three c-Jun binding sites on the MMP-20 promoter were identified for the first time, and were occupied by c-Jun as MMP-20 was induced. Deletion of any one of AP-1 binding sites on the MMP-20 promoter significantly reduced the transcription of downstream luciferase reporter. These in vitro findings suggest that c-Jun is a key regulatory element for MMP-20 expression, and human ameloblast lineage cells can respond to fluoride by down-regulating MMP-20 transcription through the JNK/c-Jun signaling pathway. PMID:17611094

  12. Expression Optimization and Characterization of the Catalytic Domain of Human MT3-MMP

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Matrix metalloproteinases(MMPs) are a family of proteases that are required for many biological processes and are also elevated in many pathological conditions. MMP inhibitors (MMPIs) may therefore be useful as therapeutic agents in treating a number of diseases including cancer, cardiovascular diseases and arthritis. Attempts have been made to develop MMPIs. Recombinant MMPs have been used to screening MMPs in vitro assays. In this work, we report the expression of MMP-16 in E. coli and the characterization of the recombinant MMP-16 with a commonly used MMP substrate DQ-gelatin.

  13. Detection of MMP-2 and MMP-9 in the Lesions of Psoriasis%银屑病皮损中MMP-2 MMP-9的检测

    Institute of Scientific and Technical Information of China (English)

    陈晋广; 任小丽; 胡雅玉; 陈祥恩

    2005-01-01

    目的探讨MMP-2,MMP-9在细胞外基质(ECM)降解和银屑病发病机理中的作用及意义.方法采用免疫组化ABC法检测银屑病患者皮损中MMP-2,MMP-9.结果银屑病皮损中既可检测到MMP-2,又可检测到MMP-9,而在正常皮肤则为阴性.结论MMP-2,MMP-9参与了银屑病的发病过程.

  14. IL-8、 MMP-9、 INF-γ的检测对结核性脑膜炎及病毒性脑膜炎发病的意义%Detection of interleukin-8, matrix metalloproteinase-9 and interferon gamma levels in the cerebrospinal fluid of patients with tuberculous meningitis and viral meningitis

    Institute of Scientific and Technical Information of China (English)

    朱飞; 张家堂; 邢小微; 贺路星; 赵威; 郎森阳; 于生元

    2012-01-01

    目的 探讨脑脊液中白细胞介素-8(IL-8)、基质金属蛋白酶-9(MMP-9)、干扰素-γ(INF-γ)含量的检测对结核性脑膜炎及病毒性脑膜炎的临床诊断价值. 方法 选取解放军总医院、解放军第三0九医院自2010年8月至2011年11月住院的患者,其中结核性脑膜炎组20例,病毒性脑膜炎组15例,非感染性神经系统疾病组20例.用ELISA法检测3组患者脑脊液IL-8、MMP-9、INF-γ含量,并进行比较分析. 结果 结核性脑膜炎组患者脑脊液中IL-8、MMP-9、INF-γ的含量高于病毒性脑膜炎组和非感染性神经系统疾病组差异有统计学意义(P<0.05).病毒性脑膜炎组患者脑脊液中IL-8、MMP-9含量高于非感染性神经系统组(P<0.05).病毒性脑膜炎组患者脑脊液中INF-γ含量与非感染性神经系统疾病组比较差异无统计学意义(P>0.05). 结论 脑脊液中IL-8、MMP-9、INF-γ含量的检测对结核性脑膜炎具有一定的辅助诊断意义.IL-8、MMP-9在病毒性脑膜炎的发病和进展中亦起到一定作用.临床上若在患者脑脊液中检测到高水平的INF-γ,较之IL-8、MMP-9对于结核性脑膜炎更具诊断价值.%Objective To investigate the diagnostic values of interleukin-8 (IL-8), matrix metalloproteinase-9 (MMP-9) and interferon gamma (INF-γ) levels in patients with tuberculous meningitis and viral meningitis by detecting the contents of these biomarkers in the cerebrospinal fluid (CSF). Methods Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of IL-8,MMP-9 and INF-γ in the CSF of patients with tuberculous meningitis (n=20),viral meningitis (n=15) and noninfectious neurologic diseases (n=20) who admitted to our hospital from August 2010 to November 2011. Results The IL-8,MMP-9 and INF-γlevels in the samples from the tuberculous meningitis patients were significantly higher than those from either viral meningitis or noninfectious neurologic diseases (P<0.05).The contents of IL-8

  15. MMP-2 -1575G/A polymorphism modifies the onset of optic neuritis as a first presenting symptom in MS?

    Science.gov (United States)

    Gašparović, Iva; Čizmarević, Nada Starčević; Lovrečić, Luca; Perković, Olivio; Lavtar, Polona; Sepčić, Juraj; Jazbec, Saša Šega; Kapović, Miljenko; Peterlin, Borut; Ristić, Smiljana

    2015-09-15

    Previous studies show altered activities of matrix metalloproteinase (MMP)-2 and MMP-9 in serum and cerebrospinal fluid of multiple sclerosis (MS) and neuromyelitis optica (NMO) patients. Optic neuritis (ON) is a common symptom of both disorders. Here we investigated the impacts of MMP-2 -1575G/A and MMP-9 -1562 C/T gene polymorphisms on disease phenotype in 100 MS patients with ON as a first symptom and 376 MS patients with other initial symptomatology. The MMP-2 -1575G/A polymorphism led to a 5-year-earlier age of disease onset in MS patients with ON as a first symptom (p=0.009).

  16. Effects of Etch-and-Rinse and Self-etch Adhesives on Dentin MMP-2 and MMP-9

    Science.gov (United States)

    Mazzoni, A.; Scaffa, P.; Carrilho, M.; Tjäderhane, L.; Di Lenarda, R.; Polimeni, A.; Tezvergil-Mutluay, A.; Tay, F.R.; Pashley, D.H.; Breschi, L.

    2013-01-01

    Auto-degradation of collagen matrices occurs within hybrid layers created by contemporary dentin bonding systems, by the slow action of host-derived matrix metalloproteinases (MMPs). This study tested the null hypothesis that there are no differences in the activities of MMP-2 and -9 after treatment with different etch-and-rinse or self-etch adhesives. Tested adhesives were: Adper Scotchbond 1XT (3M ESPE), PQ1 (Ultradent), Peak LC (Ultradent), Optibond Solo Plus (Kerr), Prime&Bond NT (Dentsply) (all 2-step etch-and-rinse adhesives), and Adper Easy Bond (3M ESPE), Tri-S (Kuraray), and Xeno-V (Dentsply) (1-step self-etch adhesives). MMP-2 and -9 activities were quantified in adhesive-treated dentin powder by means of an activity assay and gelatin zymography. MMP-2 and MMP-9 activities were found after treatment with all of the simplified etch-and-rinse and self-etch adhesives; however, the activation was adhesive-dependent. It is concluded that all two-step etch-and-rinse and the one-step self-etch adhesives tested can activate endogenous MMP-2 and MMP-9 in human dentin. These results support the role of endogenous MMPs in the degradation of hybrid layers created by these adhesives. PMID:23128110

  17. Combined spectroscopy and molecular modeling studies on the binding of galbanic acid and MMP9.

    Science.gov (United States)

    Kiani, Amir; Almasi, Khadijeh; Shokoohinia, Yalda; Sadrjavadi, Komail; Nowroozi, Amin; Shahlaei, Mohsen

    2015-11-01

    The molecular mechanism of galbanic acid (GBA) binding to matrix metalloproteinase 9 (MMP9) was investigated by fluorescence quenching, absorption spectroscopy, FT-IR, molecular docking and molecular dynamics (MD) simulation procedures. The fluorescence emission of MMP9 was quenched by GBA. The titration of MMP9 by various amount of GBA was also followed by UV-Vis absorption spectroscopy. The results revealed that GBA, as a biologically active sesquiterpene coumarin derivative, has an ability to bind strongly to MMP9. Molecular docking results indicated that the main active binding site for GBA has been located in a hydrophobic cavity in the vicinity of Zn atom. Moreover, MD simulation results suggested that GBA as a coumarin derivative can interact with MMP9, without affecting the secondary structure of MMP9. MD simulations, molecular docking as computational methods from one hand and experimental data from other hand reciprocally supported each other.

  18. Mechanical stretch induces MMP-2 release and activation in lung endothelium: role of EMMPRIN.

    Science.gov (United States)

    Haseneen, Nadia A; Vaday, Gayle G; Zucker, Stanley; Foda, Hussein D

    2003-03-01

    High-volume mechanical ventilation leads to ventilator-induced lung injury. This type of lung injury is accompanied by an increased release and activation of matrix metalloproteinases (MMPs). To investigate the mechanism leading to the increased MMP release, we systematically studied the effect of mechanical stretch on human microvascular endothelial cells isolated from the lung. We exposed cells grown on collagen 1 BioFlex plates to sinusoidal cyclic stretch at 0.5 Hz using the Flexercell system with 17-18% elongation of cells. After 4 days of cell stretching, conditioned media and cell lysate were collected and analyzed by gelatin, casein, and reverse zymograms as well as Western blotting. RT-PCR of mRNA extracted from stretched cells was performed. Our results show that 1) cyclic stretch led to increased release and activation of MMP-2 and MMP-1; 2) the activation of MMP-2 was accompanied by an increase in membrane type-1 MMP (MT1-MMP) and inhibited by a hydroxamic acid-derived inhibitor of MMPs (Prinomastat, AG3340); and 3) the MMP-2 release and activation were preceded by an increase in production of extracellular MMP inducer (EMMPRIN). These results suggest that cyclic mechanical stretch leads to MMP-2 activation through an MT1-MMP mechanism. EMMPRIN may play an important role in the release and activation of MMPs during lung injury.

  19. Serotonin-Exacerbated DSS-Induced Colitis Is Associated with Increase in MMP-3 and MMP-9 Expression in the Mouse Colon.

    Science.gov (United States)

    Chen, Menglu; Gao, Lei; Chen, Pan; Feng, Dandan; Jiang, Yalin; Chang, Yongchao; Jin, Jianjun; Chu, Fong-Fong; Gao, Qiang

    2016-01-01

    Background. 5-HT enhances dextran sulfate sodium- (DSS-) induced colitis and is involved in inflammatory bowel disease (IBD). Matrix metalloproteinases (MMPs) play roles in the process of intestinal inflammation. Aims. To examine whether 5-HT induces MMPs expression in mouse colon to enhance DSS-induced colitis. Materials and Methods. C57BL/6J (B6) mice were treated with either low-dose (1.0 mg/kg) or high-dose (2.0 mg/kg) 5-HT by enema, low-dose (1.0%) or high-dose (2.5%) DSS, or combined low-dose (1.0%) DSS and (1.0 mg/kg) 5-HT. Mouse colitis was analyzed. MMPs and tissue inhibitors of MMPs (TIMPs) mRNA were measured by real-time quantitative RT-PCR in mouse colon and in human Caco-2 cells and neutrophils. MMP-3 and MMP-9 protein levels were quantified from immunohistochemistry (IHC) images of mouse colons. Results. 5-HT exacerbated DSS-induced colitis, low-dose 5-HT induces both MMP-3 and MMP-9, and high-dose 5-HT only increased MMP-3 mRNA expression in mouse colon. Mouse colon MMP-3 and MMP-9 protein levels were also elevated by 5-HT treatment. The MMP-2, TIMP-1, and TIMP-2 mRNA levels were increased in the inflamed colon. 5-HT induced MMP-3 and MMP-9 mRNA expression in Caco-2 and human neutrophils, respectively, in vitro. Conclusion. 5-HT induced MMP-3 and MMP-9 expression in mouse colon; these elevated MMPs may contribute to DSS-induced colitis.

  20. Expression and significance of MMP-2,MMP-3 and MMP-9 in the gastric carcinoma%MMP-2、MMP-3和 MMP-9在胃癌患者中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    杨云鹏; 刘莉; 李慧

    2016-01-01

    目的:研究 MMP-2、MMP-3和 MMP-9在胃癌细胞转移中作用,进一步探讨 MMP-2、MMP-3和 MMP-9的临床意义。方法体外培养 SGC-7901胃癌细胞和 GES-1胃上皮细胞,划痕损伤实验模型和 Transwell 小室模型法检测这2种细胞的迁移, ELISA 法检测培养液中 MMP-2、MMP-3和 MMP-9的含量。免疫组化检测胃癌、胃炎患者和正常人 MMP-3的阳性率,ELISA 检测血清中 MMP-2、MMP-3和 MMP-9的含量。结果细胞划痕实验和 Transwell 实验均显示,SGC-7901细胞迁移能力强于 GES-1;SGC-7901分泌的3种蛋白含量均高于 GES-1;免疫组化结果显示,胃癌患者 MMP-3的阳性率84.4%,高于胃炎患者和正常人。其血清中 MMP-2、MMP-3和 MMP-9也明显高于胃炎患者和正常人。结论胃癌细胞的转移能力与 MMP-2、MMP-3和 MMP-9的含量相关,3种蛋白可用于胃癌的早期诊断。%Objecitive The research was in order to explore the roles of MMP-2,MMP-3 and MMP 9 which played in the gastric cancer cell metastasis,and make a further study of the clinical signifance.Methods Both SGC-7901 and GES-1 were cultured in vivo.The migra-tion were examined by Wound-healing and the Transwell assay.The expression of MMP-2,MMP-3 and MMP 9 in the medium were detected by ELISA.The positive rate of MMP-3 in the gastric cancer,gastritis and normal was used the IHC,and the serum of MMP-2,MMP-3 and MMP-9 were also examined by ELISA.Results Both Wound-healing and Transwell assay showed that the migration of SGC-7901 cells was more quicker than the GES-1.Comparing to GES-1,the MMP-2,MMP-3 and MMP-9 were higher in the SGC-7901.The immunohistochemical results showed that the positive rate of MMP-3 in the gastric cancer patients was 84.4%,it was higher than the latter.The serum levels of MMP-2 MMP-3 and MMP-9 were also significantly higher than others.Conclusion The migration of gastric cancer cells was depended on the MMP-2,MMP-3 and MMP-9.The 3 proteins can be used

  1. Genetic polymorphism of MMP family and coronary disease susceptibility: a meta-analysis.

    Science.gov (United States)

    Li, Min; Shi, Jingpu; Fu, Lingyu; Wang, Hailong; Zhou, Bo; Wu, Xiaomei

    2012-03-01

    The issue that genetic polymorphism of matrix metalloproteinase (MMP) family is in association with coronary disease is controversial. So we did a meta-analysis to clarify it clearly. We made a literature search of PubMed, the Web of Science, and Cochrane Collaboration's database to identify eligible reports. The methodological quality of each included studies was assessed. We calculated the pooled ORs with their 95%CI for each genetic polymorphism in STATA 11 software. Separate analysis was performed to address the consistency of results across the subgroup with different continents. A total of 39 studies were included, with a sample of 42269 individuals. This meta-analysis provided evidence that genetic polymorphism of MMP1-1607 1G/2G, MMP3-Gly45lys, MMP3-376 G/C, MMP3-1171 5A/6A, MMP9-1562 C/T and MMP9-R279Q have a small to medium effect on incidence of coronary disease. There was no evidence that MMP1-519 A/G, MMP1-340 T/C and MMP2-1306 C/T polymorphism could increase risk of coronary disease. Results from subgroup analysis supported a relation between MMP3-1711 5A allele, MMP9-1562 C allele and coronary disease especially in Asian population. The results provide moderate association between the six common genetic polymorphism of matrix metalloproteinase family and coronary disease. However, the challenge for researcher is identifying separate effect on different races.

  2. 大鼠分泌性中耳炎中细胞外基质重塑现象的观察

    Institute of Scientific and Technical Information of China (English)

    刘俞; 祝威; 付涛

    2012-01-01

    目的 观察大鼠分泌性中耳炎(secretory otitis media,SOM)模型中是否存在细胞外基质重塑现象及相关细胞因子的表达情况.方法 24只大鼠听泡内注入脂多糖和纤维蛋白封闭剂制作SOM模型,1、7、20 d取听泡;8只大鼠听泡内注入生理盐水为对照组.Masson染色测定中耳黏膜内胶原成分的比例;免疫组化检测转化生长因子β(transforming growth factor-β,TGFβ1)、基质金属蛋白酶2 (matrix metallo proteinases,MMP2)、金属蛋白酶组织抑制因子1 (tissue inhibitors of metallo proteinases,TIMP1)在中耳黏膜表达情况.结果 细胞外胶原随病程的延长增多;SOM组与对照组比较MMP2、TIMP1、TGF-β1蛋白在中耳黏膜表达明显增强,MMP2、TIMP1在7d时表达最强,TGF-β1在20 d时表达最强.结论 大鼠SOM模型的中耳黏膜内存在细胞外基质重塑的现象,这种现象的发生可能与TGF-β与MMPs蛋白的平衡失调有关.

  3. Overexpression of MMP13 Is Associated with Clinical Outcomes and Poor Prognosis in Oral Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Vui King Vincent-Chong

    2014-01-01

    Full Text Available Matrix metalloproteinase 13 (MMP13 plays a central role in the MMP activation cascade that enables degradation of the extracellular matrix and basement membranes, and it is identified as a potential driver in oral carcinogenesis. Therefore, this study aims to determine the copy number, mRNA, and protein expression of MMP13 in oral squamous cell carcinoma (OSCC and to associate these expressions with clinicopathological parameters. Copy number, mRNA, and protein expression analysis of MMP13 were determined using real-time quantitative PCR and immunohistochemistry methods in OSCC samples. The correlations between MMP13 expressions and clinicopathological parameters were evaluated, and the significance of MMP13 as a prognostic factor was determined. Despite discrepancies between gene amplification and mRNA and protein overexpression rates, OSCC cases showed high amplification of MMP13 and overexpression of MMP13 at both mRNA and protein levels. High level of MMP13 protein expression showed a significant correlation with lymph node metastasis (P=0.011 and tumor staging (P=0.002. Multivariate Cox regression model analysis revealed that high level of mRNA and protein expression of MMP13 were significantly associated with poor prognosis (P<0.050. Taken together, these observations indicate that the MMP13 protein overexpression could be considered as a prognostic marker of OSCC.

  4. Expression of matrix metalloprotienase-9 and extracellular matrix metalloproteinase inducers in the labial gland of patients with primary Sjiigrens's syndrome%原发性干燥综合征患者唇腺组织中MMP-9与CD147的表达及意义

    Institute of Scientific and Technical Information of China (English)

    竺红; 宫怡; 王莉; 朱蓉; 池淑红

    2008-01-01

    Objective patients with pSS and 30 cases of healthv contrels.The streptavidin immunohistochemical staining was performed to detect the expression and distribution of MMP-9 and CD147 in labial salivary glands.Quantitative analysis was performed by image analysis software-image plus 5.0 at the site of positive expression of MMP-9 and CD147.The correlation between their expression and the infiltrating lymphocyte foci per 4 mm2 of labial gland was analyzed by SPSS software as well as the correlation between the expression of MMP-9 and CD 147 in the salivary glands of patients with pSS.Results MMP-9 was hiKhly expressed in labial salivary glands from 52 patients with pSS and 30 healthy controls,but the expression of MMP-9 in pSS was stronger compared with that of healthy controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of MMP-9 in labial salivary glands from 52 patients with pSS(P<0.01).CD147 was highly expressed in labial salivary glands from 52 patients with pSS and 23 healthy controls,but over-expression of CD147 in PSS was more Drominent compared with that of controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of CD147 in labial salivary glands from 52 patients with pSS(P<0.01).The expression of MMP-9 and CDl47 was detected in ductal and acinar epithelial cells,lymphocyte foci in pSS.There was linear correlation between the expression of MMP-9 and CDl47 in the salivary glands of patients with pSS(P<0.01).Conclusion The results of this study suggest that the abnormal expression of MMP-9 and CD 147 is involved in the pathogenesis of pSS and play a crucial role.The interaction of MMP-9 and CD147 may be one of the important mechanisms leading to labial salivary glands destruction found in pSS.%目的 探讨基质金属蛋白酶(MMP)-9、细胞外基质金属蛋白酶诱导因子(CD147)在原发性干燥综合征(pSS)发病过

  5. Antisense MMP-9 RNA inhibits malignant glioma cell growth in vitro and in vivo

    Institute of Scientific and Technical Information of China (English)

    Cuiyun Sun; Qian Wang; Hongxu Zhou; Shizhu Yu; Alain R.Simard; Chunsheng Kang; Yanyan Li

    2013-01-01

    The matrix-degrading metalloproteinases (MMPs),particularly MMP-9,play important roles in the pathogenesis and development of malignant gliomas.In the present study,the oncogenic role of MMP-9 in malignant glioma cells was investigated via antisense RNA blockade in vitro and in vivo.TJ905 malignant glioma cells were transfected with pcDNA3.0 vector expressing antisense MMP-9 RNA (pcDNA-ASMMP9),which significantly decreased MMP-9 expression,and cell proliferation was assessed.For in vivo studies,U251 cells,a human malignant glioma cell line,were implanted subcutaneously into 4-to 6-week-old BALB/c nude mice.The mice bearing well-established U251 gliomas were treated with intratumoral pcDNA-AS-MMP9-Lipofectamine complex (AS-MMP-9-treated group),subcutaneous injection of endostatin (endostatin-treated group),or both (combined therapy group).Mice treated with pcDNA (empty vector)-Lipofectamine served as the control group.Four or eight weeks later,the volume and weight of tumor,MMP-9 expression,microvessel density and proliferative activity were assayed.We demonstrate that pcDNA-AS-MMP9 significantly decreased MMP-9 expression and inhibited glioma cell proliferation.Volume and weight of tumor,MMP-9 expression,microvessel density and proliferative activity in the antisense-MMP-9-treated and therapeutic alliance groups were significantly lower than those in the control group.The results suggest that MMP-9 not only promotes malignant glioma cell invasiveness,but also affects tumor cell proliferation.Blocking the expression of MMP-9 with antisense RNA substantially suppresses the malignant phenotype of glioma cells,and thus can be used as an effective therapeutic strategy for malignant gliomas.

  6. 芝麻素对代谢综合症大鼠心肌NT、NF-κB和MMP-9蛋白表达的影响%Sesamin represses protein expression of nitrotyrosine, NF-κB and matrix metalloproteinase-9 in myocardium of metabolic syndrome rats

    Institute of Scientific and Technical Information of China (English)

    孔祥; 杨解人; 张明义; 吴向起

    2011-01-01

    目的 探讨芝麻素改善代谢综合症(metabolic syndrome,MS)大鼠心肌重构的作用机制.方法 采用两肾一夹术伴高脂高糖饮食制备MS大鼠模型,灌服不同剂量芝麻素(120、60、30 mg·kg-1·d-1)8周后处死动物,称全心湿重和左心室湿重,测血清总抗氧化能力(total antioxidant capability,T-AOC),Western blot检测心肌硝基酪氨酸(nitrotyrosine,NT)、核因子-κB(NF-κB)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白表达,HE和Masson染色观察心肌病理变化并测定胶原容积分数.结果 与模型组相比,60、120 mg·kg-1芝麻素组全心湿重降低,T-AOC升高,心肌NT、NF-κB和MMP-9蛋白表达降低.120 mg·kg-1芝麻素组左心室湿重和胶原容积分数降低,心肌病理损伤明显改善.结论 芝麻素具有改善MS大鼠心肌重构的作用,其机制除抗氧化应激外,还与下调NF-κB和MMP-9蛋白表达有关.%Aim To investigate the possible in vivo protective effects of sesamin on myocardial remodeling in metabolic syndrome ( MS ) rats.Methods MS rat model was induced by operation of two-kidneys with one-clip and fed with a high-fat, high-sucrose diet.sesamin ( 120, 60 and 30 mg ·kg -1· d-1 ) was orally administered in MS rats.Eight weeks later.rats were sacrificed, heart wet weight ( HWW ) and left ventricle wet weight ( LVWW ) were recorded , total-antioxidative activity ( T-AOC ) in serum was measured, and protein expression of nitrotyrosine ( NT ), NF-KB and matrix metalloproteinase-9 ( MMP-9 ) in myocardium was detected by Western blot.In addition, pathological changes in myocardium were observed with HE and Masson staining and collagen volume fraction ( CVF ) was measured.Results Compared with MS rats, treatment with 120 and 0 mg·kg-1 sesamin decreased HWW.enhanced T-AOC, and reduced NT, NF-KB and MMP9 protein expression.Treatment with 120 mg· kg-1 sesamin decreased LVWW and CVF.and obviously ameliorated myocardial pathologic lesions

  7. [Reference ranges of matrix metalloproteinase-1, -2, -9 and tissue inhibitor of matrix metalloproteinases-1 concentrations in amniotic fluid in physiological pregnancy].

    Science.gov (United States)

    Korenovsky, Yu V; Remneva, O V

    2016-01-01

    The aim of this study was to determine reference values of matrix metalloproteinase-1 (MMP-1), MMP-2, MMP-9 and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) in the amniotic fluid at the first stage of labor in physiological pregnancy. 89 women at the first stage of term labor have been examined. Samples of amniotic fluid were taken at the first period of labor by vaginal amniotomy. Concentrations ofMMP-1, MMP-2, MMP-9, and TIMP-1 were investigated in amniotic fluid by ELISA kits. We have determined normal concentration ranges for MMP-1, MMP-2, MMP-9, TIMP-1, and ratios of concentrations of MMPs and TIMP-1 (MMP-1/TIMP-1, MMP-2/TIMP-1, MMP-9/TIMP-1) in the amniotic fluid at the first period of labor in physiological pregnancy. These included: MMP-1--5.1-16.8 pg/mg of protein, MMP-2--238.3-374.1 pg/mg of protein, MMP-9--66.1-113.3 pg/mg of protein, TIMP-1--4.7-13.6 pg/mg of protein, ratio of MMP-1/TIMP-1--0.1-2.2, ratio of MMP-2/TIMP-1--19.9-55.7, ratio of MMP-9/TIMP-1--4.2-17.2.

  8. Immunohistochemical expression of MMP-14 and MMP-2, and MMP-2 activity during human ovarian follicular development

    OpenAIRE

    2014-01-01

    BACKGROUND: The aim of this study was to investigate the presence of MMP-14 and MMP-2 during human ovarian follicular development using immunohistochemistry, and the activity of MMP-2 in follicular fluid using zymography. METHODS: Ovarian tissue collected from the archives of the Department of Pathology was examined and medical records and histopathology were reviewed. Follicular fluids were collected at the IVF-department and analyzed using zymography. RESULTS: MMP-14 and MMP-2 were increasi...

  9. Inhibition of pro-/active MMP-2 by green tea catechins and prediction of their interaction by molecular docking studies.

    Science.gov (United States)

    Chowdhury, Animesh; Nandy, Suman Kumar; Sarkar, Jaganmay; Chakraborti, Tapati; Chakraborti, Sajal

    2017-03-01

    Matrix metalloproteinases (MMPs) play a crucial role in developing different types of lung diseases, e.g., pulmonary arterial hypertension (PAH). Green tea polyphenolic catechins such as EGCG and ECG have been shown to ameliorate various types of diseases including PAH. Our present study revealed that among the four green tea catechins (EGCG, ECG, EC, and EGC), EGCG and ECG inhibit pro-/active MMP-2 activities in pulmonary artery smooth muscle cell (PASMC) culture supernatant. Based on the above, we investigated the interactions of pro-/active MMP-2 with the green tea catechins by computational methods. In silico analysis revealed a strong interaction of pro-/active MMP-2 with EGCG/ECG, and galloyl group has been observed to be responsible for this interaction. The in silico analysis corroborated our experimental observation that EGCG and ECG are active in preventing both the proMMP-2 and MMP-2 activities. Importantly, these two catechins appeared to be better inhibitors for proMMP-2 in comparison to MMP-2 as revealed by gelatin zymogram and also by molecular docking studies. In many type of cells, activation of proMMP-2 occurs via an increase in the level of MT1-MMP (MMP-14). We, therefore, determined the interactions of MT1-MMP with the green tea catechins by molecular docking analysis. The study revealed a strong interaction of MT1-MMP with EGCG/ECG, and galloyl group has been observed to be responsible for the interaction.

  10. MMP-1 serum levels predict coronary atherosclerosis in humans

    Directory of Open Access Journals (Sweden)

    Reiser Maximilian

    2009-09-01

    Full Text Available Background Myocardial infarction results as a consequence of atherosclerotic plaque rupture, with plaque stability largely depending on the lesion forming extracellular matrix components. Lipid enriched non-calcified lesions are considered more instable and rupture prone than calcified lesions. Matrix metalloproteinases (MMPs are extracellular matrix degrading enzymes with plaque destabilisating characteristics which have been implicated in atherogenesis. We therefore hypothesised MMP-1 and MMP-9 serum levels to be associated with non-calcified lesions as determined by CT-angiography in patients with coronary artery disease. Methods 260 patients with typical or atypical chest pain underwent dual-source multi-slice CT-angiography (0.6-mm collimation, 330-ms gantry rotation time to exclude coronary artery stenosis. Atherosclerotic plaques were classified as calcified, mixed or non-calcified. Results In multivariable regession analysis, MMP-1 serum levels were associated with total plaque burden (OR: 1.37 (CI: 1.02-1.85; p Conclusion MMP-1 serum levels are associated with total plaque burden but do not allow a specification of plaque morphology.

  11. 苦参碱对体外培养人增生性瘢痕成纤维细胞 MMP-1、MMP-9表达的影响%Effects of Matrine on Expressions of MMP-1,MMP-9 in Human Hypertropic Scar Fibroblasts in vitro

    Institute of Scientific and Technical Information of China (English)

    陈小婷; 欧斌贤; 唐屈; 黄积荣; 刘达恩; 农庆文

    2014-01-01

    Objective To study the effects of matrine on the expressions of matrix metalloproteinase (MMP)-1, MMP-9 in human hypertropic scar fibroblasts ( HSFb) in vitro.Methods HSFb was cultured in vitro by being treated with different concentrations of matrine[0 mmol/L(control group),2.5 mmol/L,5.0 mmol/L,10.0 mmol/L].Enzyme-linked immunoadsorbent assay(ELISA) was used to detect the expressions of MMP-1,MMP-9 in the treated supernatants.Results The expression levels of MMP-1,MMP-9 in groups of different concentrations of matrine were higher than those in the control group 24 or 48 hours after matrine treatment(P<0.05),the expression levels of MMP-1,MMP-9 in the 5.0 mmol/L matrine group were higher than those in the 2.5,10.0 mmol/L matrine groups ( P<0.05).Conclusion Matrine can increase the expression of MMP-1,MMP-9.It can be applied to hyperplastic scar prevention and treatment .%目的:探讨苦参碱对体外培养人增生性瘢痕成纤维细胞( HSFb )基质金属蛋白酶( MMP-1、MMP-9)表达的影响。方法将浓度分别为0 mmol/L(对照组)、2.5 mmol/L、5.0 mmol/L、10.0 mmol/L苦参碱作用于体外培养人HSFb,采用酶联免疫吸附试验检测加药后上清液中MMP-1、MMP-9表达量。结果苦参碱作用24 h、48 h时各浓度组MMP-1、MMP-9的表达量均高于对照组(P<0.05),5.0 mmol/L组表达量明显高于2.5 mmol/L组、10.0 mmol/L组( P<0.05)。结论苦参碱可增加HSFb MMP-1、MMP-9的表达,抗瘢痕作用好,在增生性瘢痕防治上具有一定的应用前景。

  12. MMP-2 Alters VEGF Expression via αVβ3 Integrin-Mediated PI3K/AKT Signaling in A549 Lung Cancer Cells

    OpenAIRE

    2010-01-01

    Vascular endothelial growth factor (VEGF) is one of the most important angiogenic growth factors for tumor angiogenesis. Here, we sought to explore whether RNA interference (RNAi) targeting Matrix metalloproteinase-2 (MMP-2) could disrupt VEGF mediated angiogenesis in lung cancer. MMP-2 siRNA inhibited lung cancer cell-induced tube formation of endothelial cells in vitro; addition of recombinant human-MMP-2 restored angiogenesis. MMP-2 transcriptional suppression decreased VEGF, PI3K protein ...

  13. MMP-2 detective silicon nanowire biosensor using enzymatic cleavage reaction.

    Science.gov (United States)

    Choi, Jin-Ha; Kim, Han; Kim, Hyun-Soo; Um, Soong Ho; Choi, Jeong-Woo; Oh, Byung-Keun

    2013-04-01

    Matrix metalloproteinases are proteolytic enzymes that play a significant role in tissue remodeling related with various pathological and physiological processes such as tissue repair, angiogenesis, cirrhosis, morphogenesis, arthritis, and metastasis. Especially, MMP-2 has been shown to be related with benign prostatic hyperplasia and prostate cancer. Therefore, there is a need to make sensors with high sensitivity that can measure MMP-2 concentrations precisely. Silicon nanowires have been used in the development of high sensitive chemical sensors and biosensors. The high sensitivity of silicon nanowire based sensor originates in its high surface to volume ratio and ability to field-effect induced local charge transfers. In this study, 100 nm silicon nanowire based field-effect transistors (FET) device was fabricated by electron-beam lithography and MMP-2 was successfully measured by conductance versus time characteristics within 1 pM to 100 nM.

  14. Matrix Metalloproteinases Expression in Choroidal Neovascular Membranes

    Institute of Scientific and Technical Information of China (English)

    Jun Zeng; Deyong Jiang; Xiangping Liu; Xiaohua Zhu; Luosheng Tang

    2004-01-01

    Purpose: To investigate the expression of matrix metalloproteinases (MMPs) in choroidal neovascular membranes with age-related macular degeneration (AMD).Methods: Seventeen choroidal neovascular membranes surgically removed from AMD patients with pars plana vitrectomy and subretinal membranes peeling were investigated.The expression of MMP-2 and MMP-9 was determined with immunohistochemical technique.Results: Immunohistochemistry staining in choroidal neovascular membranes for MMP2 and MMP-9 was observed in 17 specimens. There was no detective of MMP-2 and MMP-9 in normal retinas.Conclusions: MMP-2 and MMP-9 were found in choroidal neovascular membranes, may degrade the Bruch membrane and be associated with the perforation of new vessels into Bruch membrane, involving a basic pathogenic process of AMD.

  15. Recognition of Streptococcus pneumoniae and muramyl dipeptide by NOD2 results in potent induction of MMP-9, which can be controlled by lipopolysaccharide stimulation.

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    Vissers, Marloes; Hartman, Yvonne; Groh, Laszlo; de Jong, Dirk J; de Jonge, Marien I; Ferwerda, Gerben

    2014-12-01

    Matrix metallopeptidase 9 (MMP-9) is a protease involved in the degradation of extracellular matrix collagen. Evidence suggests that MMP-9 is involved in pathogenesis during Streptococcus pneumoniae infection. However, not much is known about the induction of MMP-9 and the regulatory processes involved. We show here that the Gram-positive bacteria used in this study induced large amounts of MMP-9, in contrast to the Gram-negative bacteria that were used. An important pathogen-associated molecular pattern (PAMP) for Gram-positive bacteria is muramyl dipeptide (MDP). MDP is a very potent inducer of MMP-9 and showed a dose-dependent MMP-9 induction. Experiments using peripheral blood mononuclear cells (PBMCs) from Crohn's disease patients with nonfunctional NOD2 showed that MMP-9 induction by Streptococcus pneumoniae and MDP is NOD2 dependent. Increasing amounts of lipopolysaccharide (LPS), an important PAMP for Gram-negative bacteria, resulted in decreasing amounts of MMP-9. Moreover, the induction of MMP-9 by MDP could be counteracted by simultaneously adding LPS. The inhibition of MMP-9 expression by LPS was found to be regulated posttranscriptionally, independently of tissue inhibitor of metalloproteinase 1 (TIMP-1), an endogenous inhibitor of MMP-9. Collectively, these data show that Streptococcus pneumoniae is able to induce large amounts of MMP-9. These high MMP-9 levels are potentially involved in Streptococcus pneumoniae pathogenesis.

  16. Dieckol from Ecklonia cava Regulates Invasion of Human Fibrosarcoma Cells and Modulates MMP-2 and MMP-9 Expression via NF-κB Pathway

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    Chen Zhang

    2011-01-01

    Full Text Available The matrix metalloproteinase (MMP family is involved in the breakdown of extracellular matrix in normal physiological processes, as well as in the disease processes such as arthritis and cancer metastasis. In the present study, dieckol was obtained with high yield from marine brown alga Ecklonia cava (EC, and its effect was assessed on the expression of MMP-2 and -9 and morphological changes in human fibrosarcoma cell line (HT1080. Dieckol inhibited the expression of MMP-2 and -9 in a dose-dependent manner and also suppressed the cell invasion and the cytomorphology in 3D culture system on HT1080 cells. Moreover, dieckol may influence nuclear factor kappa B (NF-κB pathway without obvious influence on activator protein-1 (AP-1 pathway and tissue inhibitor of metalloproteinases (TIMPs. In conclusion, dieckol could significantly suppress MMP-2 and -9 expression and alter cytomorphology of HT1080 cell line via NF-κB pathway.

  17. Local Inflammation Alters MMP-2 and MMP-9 Gelatinase Expression Associated with the Severity of Nifedipine-Induced Gingival Overgrowth: a Rat Model Study.

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    Li, Wu-Li; Wu, Cheng-Hai; Yang, Jun; Tang, Min; Chen, Long-Jie; Zhao, Shou-Liang

    2015-08-01

    Nifedipine-induced gingival overgrowth (NIGO) is characterized by cell proliferation and extracellular matrix (ECM) component accumulation in gingival connective tissues, with varying degrees of inflammation and fibrosis. Impaired collagen and ECM homeostasis may be among the underlying molecular mechanisms that lead to the fibrotic changes that occur in drug-induced gingival overgrowth (DIGO). Because matrix metalloproteinases (MMPs) play vital roles in regulating collagen and ECM metabolism, many studies have been performed to reveal the relationship between MMPs and DIGO. It is thought that the gelatinases MMP-2 and MMP-9, both type IV collagenases, are involved in the development of tissue inflammation and organ fibrosis. However, the few studies regarding gelatinase expression in DIGO are controversial. Recent studies have demonstrated the inhibitory effect of cyclosporine A (CsA) on gelatinase expression and/or activity; however, similar changes have yet to be detected in Nif-treated gingival tissues. In this study, we verified that Nif treatment could lead to gingival overgrowth in rats and that gingival inflammation played a pro-proliferative role in NIGO development. Additionally, we examined the temporal expression of gelatinases on days 0, 7, 14, 21, 30, and 40 during NIGO development. The aim was to investigate whether MMP-2 and MMP-9 played significant roles in regulating NIGO development and progression. MMP-2 gene expression was not altered by Nif treatment alone but was significantly inhibited by Nif treatment for 30 days in the presence of local inflammation. However, no significant alterations in MMP-2 protein expression were detected in the Nif-treated gingival tissue, regardless of the presence or absence of local inflammation. Moreover, Nif treatment could lead to transient and significant increases in MMP-9 gene and protein expression levels in the presence of local inflammation. In particular, active MMP-9 expression increased significantly

  18. MMP-2 is localized to the mitochondria-associated membrane of the heart.

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    Hughes, Bryan G; Fan, Xiaohu; Cho, Woo Jung; Schulz, Richard

    2014-03-01

    Matrix metalloproteinase-2 (MMP-2) has been extensively studied in the context of extracellular matrix remodeling but is also localized within cells and can be activated by prooxidants to proteolyze specific intercellular targets. Although there are reports of MMP-2 in mitochondria, a critical source of cellular oxidative stress, these studies did not take into account the presence within their preparations of the mitochondria-associated membrane (MAM), a subdomain of the endoplasmic reticulum (ER). We hypothesized that MMP-2 is situated in the MAM and therefore investigated its subcellular distribution between mitochondria and the MAM. Immunogold electron microscopy revealed MMP-2 localized in mitochondria of heart sections from mice. In contrast, immunofluorescence analysis of an MMP-2:HaloTag fusion protein expressed in HL-1 cardiomyocytes showed an ER-like distribution, with greater colocalization with an ER marker (protein disulfide isomerase) relative to the mitochondrial marker, MitoTracker red. Although MMP-2 protein and enzymatic activity were present in crude mitochondrial fractions, once these were separated into purified mitochondria and MAM, MMP-2 was principally associated with the latter. Thus, although mitochondria may contain minimal levels of MMP-2, the majority of MMP-2 previously identified as "mitochondrial" is in fact associated with the MAM. We also found that calreticulin, an ER- and MAM-resident Ca(2+) handling protein and chaperone, could be proteolyzed by MMP-2 in vitro. MAM-localized MMP-2 could therefore potentially impact mitochondrial function by affecting ER-mitochondrial Ca(2+) signaling via its proteolysis of calreticulin.

  19. Regulation of MMP-3 expression and secretion by the chemokine eotaxin-1 in human chondrocytes

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    Chao Pin-Zhir

    2011-11-01

    Full Text Available Abstract Background Osteoarthritis (OA is characterized by the degradation of articular cartilage, marked by the breakdown of matrix proteins. Studies demonstrated the involvement of chemokines in this process, and some may potentially serve as diagnostic markers and therapeutic targets; however, the underlying signal transductions are not well understood. Methods We investigated the effects of the CC chemokine eotaxin-1 (CCL11 on the matrix metalloproteinase (MMP expression and secretion in the human chondrocyte cell line SW1353 and primary chondrocytes. Results Eotaxin-1 significantly induced MMP-3 mRNA expression in a dose-dependent manner. Inhibitors of extracellular signal-regulated kinase (ERK and p38 kinase were able to repress eotaxin-1-induced MMP-3 expression. On the contrary, Rp-adenosine-3',5'-cyclic monophosphorothioate (Rp-cAMPs, a competitive cAMP antagonist for cAMP receptors, and H-89, a protein kinase A (PKA inhibitor, markedly enhanced eotaxin-1-induced MMP-3 expression. These results suggest that MMP-3 expression is specifically mediated by the G protein-coupled eotaxin-1 receptor activities. Interestingly, little amount of MMP-3 protein was detected in the cell lysates of eotaxin-1-treated SW1353 cells, and most of MMP-3 protein was in the culture media. Furthermore we found that the eotaxin-1-dependent MMP-3 protein secretion was regulated by phospholipase C (PLC-protein kinase C (PKC cascade and c-Jun N-terminal kinase (JNK/mitogen-activated protein (MAP kinase pathways. These data indicate a specific regulation of MMP-3 secretion also by eotaxin-1 receptor activities. Conclusions Eotaxin-1 not only induces MMP-3 gene expression but also promotes MMP-3 protein secretion through G protein-coupled eotaxin-1 receptor activities. Chemokines, such as eotaxin-1, could be a potential candidate in the diagnosis and treatment of arthritis.

  20. Elevated serum levels of MMP-11 correlate with poor prognosis in colon cancer patients.

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    Pang, Li; Wang, Da-Wei; Zhang, Nan; Xu, Da-Hai; Meng, Xiang-Wei

    2016-03-11

    Matrix metalloproteinase 11 (MMP11) has been shown to play a key role in human tumor progression and indicates poor clinical outcome in cancer patients. The current study aimed to evaluate the relationship between serum levels of MMP-11 and prognosis in colon cancer patients. Serum levels of MMP-11 were determined in 92 colon cancer patients and 92 healthy individuals using an enzyme-linked immunosorbent assay (ELISA). Associations between serum MMP-11 levels and clinicopathological characteristics of the patients and their outcomes were investigated. Survival analyses were performed to measure the 5-year overall survival (OS) and disease-free survival (DFS). Serum MMP-11 levels were substantially higher in colon cancer patients than in healthy controls. Moreover, serum MMP-11 levels were significantly higher in patients with advanced T status, lymph node metastasis, distant metastasis, and a higher TNM stage. Elevated serum levels of MMP-11 were identified as an independent prognostic factor for 5-year mortality and adverse events associated with colon cancer. Multivariate Cox regression analysis identified the serum MMP-11 level as an independent predictor of OS and DFS. Our study established that high serum levels of MMP-11 are associated with poor clinical outcome and may serve as a prognostic biomarker in colon cancer patients.

  1. MMP-13, p53 in the Progression of Malignant Peripheral Nerve Sheath Tumors

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    Nikola Holtkamp

    2007-08-01

    Full Text Available Malignant peripheral nerve sheath tumors (MPNST are sarcomas with poor prognosis, limited treatment options. Factors contributing to tumor progression are largely unknown. We therefore examined MPNST from 22 neurofibromatosis type 1 (NF1 patients, 14 nonNF1 patients, 14 neurofibroma patients for matrix metalloproteinase 13 (MMP-13 expression. Because wild-type, mutant p53 were shown to differentially regulate MMP-13 expression, TP53 status, protein levels were also determined. MMP-13 expression was detected in 58% of MPNST, was significantly associated with recurrent MPNST (P = .019. p53 was observed in 78% of MPNST, was found to be strongly associated with MMP-13 expression (P = .005. In contrast, 14 neurofibromas lacked MMP-13, p53 expressions. TP53 mutations were found in only 11% of MPNST, were associated with high tumor grades (P = .029. No significant association between mutant TP53, MMP-13 was observed, indicating that other factors drive MMP-13 expression in MPNST. The presence of metastasis was linked to p53Pro72 polymorphism (P= .041, shorter survival. In summary, our data suggest that MMP-13 expression in nerve sheath tumors is coupled with malignant progression. Therefore, MMP-13 may serve as a marker for progression, as a therapeutic target.

  2. EGFR signaling downstream of EGF regulates migration, invasion, and MMP secretion of immortalized cells derived from human ameloblastoma.

    Science.gov (United States)

    da Rosa, Marina Rolo Pinheiro; Falcão, Aline Semblano Carreira; Fuzii, Hellen Thais; da Silva Kataoka, Maria Sueli; Ribeiro, André L R; Boccardo, Enrique; de Siqueira, Adriane Sousa; Jaeger, Ruy G; de Jesus Viana Pinheiro, João; de Melo Alves Júnior, Sérgio

    2014-11-01

    Ameloblastoma is an odontogenic tumor characterized by local invasiveness and frequent recurrence. The surrounding stroma, composed of different cell types and extracellular matrix (ECM), may influence ameloblastoma invasive behavior. Furthermore, tumor and stromal cells secrete matrix metalloproteases (MMPs), which, in turn, can modulate the matrix and promote the release of ECM-bound growth factors. Among these growth factors, epidermal growth factor (EGF) and its receptor, EGFR, have already been shown to stimulate MMP synthesis, suggesting that an interdependent mechanism, involving MMP activity and growth factors release, may contribute to tumor invasiveness. The aim of this study was to evaluate the effects of the EGF/EGFR signaling pathway on migration, invasion, and MMP activity, in a primary cell line derived from human ameloblastoma. We established and characterized a primary cell line (AME-1) from a human ameloblastoma sample. This cell line was transduced with human papillomavirus type 16 (HPV16) E6/E7 oncogenes, generating the AME-HPV continuous cell line. EGF, MMP2, and MMP9 expression in ameloblastoma biopsies and in the AME-HPV cell line was analyzed by immunohistochemistry and immunofluorescence, respectively. Migratory activity of EGF-treated AME-HPV cells was investigated using monolayer wound assays and Transwell chambers. EGF-induced invasion was assessed in Boyden chambers coated with Matrigel. Conditioned medium from EGF-treated cells was subjected to zymography. EGFR expression in AME-HPV cells was silenced by small interfering RNA (siRNA), to verify the relationship between this receptor and MMP secretion. Ameloblastoma samples and AME-HPV cells expressed EGF, EGFR, MMP2, and MMP9. AME-HPV cells treated with EGF showed increased rates of migration and invasion, as well as enhanced MMP2 and MMP9 activity. EGFR knockdown decreased MMP2 and MMP9 levels in AME-HPV cells. EGFR signaling downstream of EGF probably regulates migration, invasion

  3. Effect of spearmint oil on lipopolysaccharide induced emphysema-like changes and expression of matrix metalloproteinase-9%留兰香油对脂多糖引起的大鼠肺气肿样改变及对MMP-9表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘君波; 王砚; 唐法娣; 余晨曦; 黄梦珊; 赵小京; 朱有法

    2011-01-01

    Objective: To investigate the effect of spearmint oil on emphysema-like changes and the expression of tumor necrosis factor-α( TNF-α), interleukin-lβ( IL-lβ), matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP9) in lipopolysaccharide (LPS) treated rats. Method: Emphysematous changes model was induced by intratracheal instillation of LPS once a week for up to 8 weeks in rats. Rats were divided into control, dexamethasone(0. 3 mg · kg-1 ), and spearmint oil( 10,30,100 mg · kg-1 ) groups. Each group was treated with saline, dexamethasone, and spearmint of oil respectively for 4 weeks. Then total and different white blood cell counts in bronchoalveolar lavage fluid(BALF) were carried out. The pathologic changes of lung tissue such as alveolar structure, airway inflammation, and goblet cell metaplasia were observed by HE and AB-PAS staining Expression of TNF-α,HL-lβ, TIMP-1 and MMP-9 were measured. Result: Both spearmint and dexamethasone decreased the destruction of pulmonary alveolts. The total and different white blood cell counts in BALF including neutrophile and lymphocyte of spearmint oil 100 mg· kg - 1 and dexamethasone group were significantly reduced, and the goblet cell metaplasia was also inhibited. Dexamethasone had inhibitory effect on the expression of TNF-α, IL-1β, TIMP-1 and MMP-9. Spearmint oil 30,100 mg · kg-1 significanfiy reduced TNF-α and IL-1β respectively. Spearmint oil 10,30 and 100 mg · kg-1 had no effect on the expression of TIMP-1, but could decrease the expression of MMP-9 significantly in lung tissues. Conclusion: Spearmint oil has protective effect on rats with emphysematous changes, since it improves alveolar destruction, pulmonary inflammation, and goblet cell metaplasia. The mechanism may include reducing TNF-α, IL-lβ content and inhibiting overexpression of matrix metalloproteinase-9 in lung tissues.%目的:观察留兰香油对脂多糖(LPS)所致大鼠肺气肿样改变及其对肿瘤

  4. Association between the matrix metalloproteinase-12 gene polymorphism and susceptibility of non-small cell lung cancer in northern China%MMP-12基因多态性与中国北方人非小细胞肺癌易感性的关系

    Institute of Scientific and Technical Information of China (English)

    王伟; 曾辉; 王瑞

    2013-01-01

    目的:探讨MMP-12启动子区-82bpA/G单核苷酸多态性(SNP)与我国北方人非小细胞肺癌(NSCLC)遗传易感性的关系.方法:采用基于医院的病例-对照研究方法,收集300例非小细胞肺癌患者(其中鳞癌161例,腺癌139例)和300例健康对照个体的静脉抗凝血5ml,以蛋白酶K消化-饱和氯化钠盐析法提取外周血白细胞DNA,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法对基质金属蛋白酶MMP-12-82A/G单核苷酸多态性进行基因分型,比较非小细胞肺癌病例组与健康对照组之间等位基因及基因型分布.结果:非小细胞肺癌病例组中吸烟个体的比例(62.0%)明显高于健康对照组(36.3%),P<0.01,经年龄、性别校正后的OR值为3.74(95% CI =2.451 ~5.705).健康对照组中的MMP-12基因启动子区转录起始点上游82 bp处A/G SNP的基因型分布符合Hardy-Weinberg平衡(P>0.05).在病例组和健康对照组之间及根据吸烟状况和病理类型分层分析MMP-12基因-82A/G SNP基因型和等位基因频率分布均无显著性差异(P>0.05).结论:吸烟可以显著增加非小细胞肺癌的发病风险.MMP-12基因-82A/G SNP与非小细胞肺癌的发病风险无关.%Objective:To investigate the association between matrix metalloproteinases-12 (MMP-12-82A/G SNP) polymorphism and susceptibiliy of non-small cell lung cancer(NSCLC) in northern China.Methods:This hospital-based case-control study included 300 patients with non-small cell lung cancer (161 squamous carcinoma and 139 adenocarcinoma) and 300 healthy controls.Total DNA was extracted form peripheral blood leucocyte using proteinase K digestion followed by a salting out procedure.Polymorphisms of MMP-12 gene were analyzed by PCR-restriction fragment length polymorphism (RFLP).Results:The percentage of smokers in the patients (62.0%)was significantly higher than that in healthy controls (36.3 %,P < 0.01).Smoking may increase the risk of NSCLC (age and

  5. Association of matrix metalioproteinase-3 serum level and the promoter 5A/6A polymorphism of the MMP-3 gene with atherosderotic cerebral infarction%基质金属蛋白酶-3血清水平及启动子基因5A/6A多态性与动脉粥样硬化性脑梗死的关系

    Institute of Scientific and Technical Information of China (English)

    黄小亚; 朱敏; 金笑平; 张丹红; 王灵芝; 叶祖森

    2008-01-01

    Objective To investigate the association of matrix metalluproteinase-3 (MMP-3) serum level and the promoter 5A/6A polymorphism of theMMP-3 gene with atherosclerotic cerebral infarction (ACI) in a Chinese Hart population.Methods Two hundred and fifteen patients with acute ACI from the Department of Neurology of Taizhou Hospi tal and 226 healthy controls were included in the study.Serum MMP-3 level was measured by enzyme-linked immunosorbent assay (ELISA).Genotype was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for the common 5A/6A functional promoter polymorphism of the MMP-3 gene.Results The genotype distribution of the MMP-3 promoter 5A/6A polymorphism between the ACI patients group and the control group was significantly different(X2=9.389,P=0.002).The frequencies of the 5A allele were 14.2%and 7.7%in the ACI patients group and the control group respectively(X2=9.430,P=0.002).Serum level of MMP-3 in the ACI patients group was significantly higher than that in the control group(t=24.867,P=0.000).Among the ACI patients group,serum MMP3 levels also had significant difference between the 5A/6A + 5A/SA and the 6A/6A genotype(t=2.057,P=0.041).Conclusion The present findings suggest that serum level of MMP-3 obviously increased within 48 hours of ischemic stroke and the genetic polymorphism of 5A/6A in the MMP-3 promoter is associated with ACI and MMP-3 expression in the Chinese Han population.%目的 探讨摹质金属蛋白酶-3(matrix metalloproteinase-3,MMP-3)血清水平及启动子基因5A/6A多态性与动脉粥样硬化性脑梗死(atherosclerotic cerebral infarction,ACI)的关系.方法 对215例ACI患者和226名健康体检者进行研究.采用ELISA法测定血清MMP-3水平,同时采用聚合酶链反应-限制性片段长度多态性方法分析MMP-3启动子基因5A/6A多态性.结果 ACI组发病48 h内的血清NIVIP-3水平为(23.28±12.19)ng/μL,较对照组(2.76±2.25)ng/μL显著增高(t=24.867,P=0

  6. TGF-β1 promoted MMP-2 mediated wound healing of anterior cruciate ligament fibroblasts through NF-κB.

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    Wang, Yequan; Tang, Zhenyu; Xue, Ruyue; Singh, Gurinder K; Lv, Yonggang; Shi, Kunning; Cai, Kaiyong; Deng, Linhong; Yang, Li

    2011-06-01

    The adult human anterior cruciate ligament (ACL) has poor functional healing response. Transforming growth factor (TGF)-β1 enhances the wound repair by stimulating matrix proteins deposition as well as the proliferation and migration of cells. However, the function of the TGF-β1-induced matrix metalloproteinases' (MMPs) activities in the wound healing process is poorly understood. In this study, exogenous MMP-2 is added to mimic the TGF-β1-induced MMP-2 expression. Role of NF-κB pathway is further examined. Our results show that TGF-β1 induces dramatic elevation of MMP-2 activities and the MMP-2/tissue inhibitors of metalloproteinases ratio. Furthermore, the exogenous MMP-2 significantly promoted in vitro wound healing abilities of ACL fibroblasts that are significantly blocked with the addition of its inhibitors. TGF-β1 also increases the proliferation of ACL fibroblasts whereas MMP-2 alone does not, indicating that MMP-2 activities are not involved in the proliferation. TGF-β1-induced MMP-2 activity is inhibited by Bay11-7082 and Bay11-7085 (NF-κB inhibitors). Our results demonstrate that increased TGF-β1 facilitates the ACL healing process by promoting the fibroblasts migration and proliferation. The migration process is mediated by MMP-2 and NF-κB pathway is involved in TGF-β1-mediated MMP-2 release.

  7. CD44 and MMP-2 expression in urothelial carcinoma

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    Gülgün ERDOĞAN

    2008-09-01

    Full Text Available Aim: CD44, one of the adhesion molecules, is thought to play an important role in cell-cell and cell-matrix interactions. Matrix metalloproteinases are degradative enzymes that remodel extracellular components. In this study the relation of MMP-2 and CD44 expressions with the histologic classification and the pathologic stage of urothelial carcinoma was revealed using immunohistochemistry.Material and Methods: Thirty-nine patients with urothelial carcinoma of the bladder were studied. The histological classification was performed according to WHO criteria. Patients were grouped as infiltrating urothelial carcinoma, low grade non-invasive papillary urothelial carcinoma, and high grade non-invasive papillary urothelial carcinoma. The pathological staging was done according to the TNM classification. Immunohistochemical staining using CD44 and MMP-2 antibodies was performed on tissue blocks.Results: CD44 immunoreactivity was detected in 77% (30/39 of the tumours which was significantly higher in non-invasive papillary urothelial carcinomas, low grade non-invasive papillary urothelial carcinomas, high grade infiltrating urothelial carcinomas (p≥0.05. MMP-2 expression was observed in 69% (27 of 39 of the tumours. There were no significant differences in MMP-2 expression between various histologic subtypes and noninvasive and infiltrative tumours.Conclusion: In conclusion, higher expression of CD44 is inversely correlated with infiltrative potential of urothelial carcinoma. These results should be supported by further studies.

  8. Novel MMP20 and KLK4 Mutations in Amelogenesis Imperfecta.

    Science.gov (United States)

    Seymen, F; Park, J-C; Lee, K-E; Lee, H-K; Lee, D-S; Koruyucu, M; Gencay, K; Bayram, M; Tuna, E B; Lee, Z H; Kim, Y-J; Kim, J-W

    2015-08-01

    In order to achieve highly mineralized tooth enamel, enamel proteinases serve the important function of removing the remaining organic matrix in the mineralization and maturation of the enamel matrix. Mutations in the kallikrein 4 (KLK4), enamelysin (MMP20), and WDR72 genes have been identified as causing hypomaturation enamel defects in an autosomal-recessive hereditary pattern. In this report, 2 consanguineous families with a hypomaturation-type enamel defect were recruited, and mutational analysis was performed to determine the molecular genetic etiology of the disease. Whole exome sequencing and autozygosity mapping identified novel homozygous mutations in the KLK4 (c.620_621delCT, p.Ser207Trpfs*38) and MMP20 (c.1054G>A, p.Glu352Lys) genes. Further analysis on the effect of the mutations on the translation, secretion, and function of KLK4 and MMP20 revealed that mutant KLK4 was degraded intracellularly and became inactive while mutant MMP20 was expressed at a normal level but secreted only minimally with proteolytic function.

  9. MMP-2 Is Mainly Expressed in Arterioles and Contributes to Cerebral Vascular Remodeling Associated with TGF-β1 Signaling.

    Science.gov (United States)

    Hua, Ye; Zhang, Weifeng; Xie, Zhenying; Xu, Nanfei; Lu, Yunnan

    2016-07-01

    There is increasing evidence to suggest that matrix metalloproteinases (MMPs) play a crucial role in vascular remodeling. It has been reported that hypoxia stimulated MMP-9 expression in brain endothelial cells and MMP-9 plays an important role in cerebral vascular remodeling. However, little is known about MMP-2 in the cerebral vessels remodeling. Herein, the aim of this study is to examine the class of vessel and cell type expressing MMP-2 in cerebral vessels and to investigate its potential role in vascular remodeling. In the present study, dual-immunofluorescence assay showed that MMP-2 was mainly expressed in arterioles. In addition, we found that MMP-2 expression in cerebral vessels was derived from endothelial cells, not astrocyte cells. Notably, in the normoxic central nervous system (CNS), there was no effect on vascular development, integrity, or endothelial proliferation when MMP-2 was knocked out, but lack of MMP-2 led to defective arteriolar remodeling and associated with transforming growth factor β1 (TGF-β1) signaling in CNS. Moreover, blocking TGF-β with SB431542, a specific TGF-β inhibitor, significantly reduced the messenger RNA (mRNA) and protein expression levels of MMP-2 in human umbilical vein endothelial cells (HUVECs). Our findings reveal that the level of MMP-2 is high in arteriolar endothelial cells and demonstrate a novel connection between MMP-2 and TGF-β1 signaling in cerebral vascular remodeling.

  10. ProMMP-1 PRODUCTION BY CULTIVATED CELLS OF VASCULAR ENDОTHELIUM IN VITRO AND IN A HUMAN BODY

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    N. N. Scliankina

    2011-01-01

    Full Text Available Abstract. Matrix metalloproteinases (MMP are structurally related endopeptidase composed of active sites which include ions Zn2+ and Ca2+. Cultured cells of human blood vessels produce MMP-1, proteolytic effect is aimed at splitting the collagen I and III types, and subsequent vascular remodeling. MMP-1 is synthesized as an inactive zymogen proMMP-1. It was shown that interferon alpha, beta and gamma inhibited production by culture of HUVEC proenzyme MMP-1, which seems to characterize their anti-angiogenic action. The effect of immunomodulators is more difficult to explain: perhaps inhibiting effect of imunofan and, as well as activating effect of cycloferon due to their internal structural peculiarities. The action of interferon alpha, beta and gamma, used as HUVEC before infection with HSV-1, and after it led to decrease in production proMMP-1. Apparently, the antiangiogenic effect of IFN is saved in the case of infection of cultures of vascular endothelium with HSV-1. Scatter in the content of proMMP-1 in the serum of blood donors was 1.625–11.8 ng/ml and in patients with chronic microbial-viral infections was 1.22–21.16 ng/ml. Higher rates of proMMP-1 were in older patients group. To estimate the system of MMP in vitro, and in the body a comprehensive study must be conducted, including proMMP-1, the active form of proenzyme and specific inhibitor of MMP-1.

  11. Expression changes and roles of matrix metalloproteinases in a rat model of traumatic deep vein thrombosis

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yu-bing; LI Wen; YAO Li-qing; ZHAO Xue-ling; WANG Bing; LI Hong-kun; NING Ya; SONG En; ZHANG Xin-xin

    2010-01-01

    Objective:To study the expression changes of matrix metal loproteinases (MMPs) in traumatic deep vein thrombosis (TDVT) in a rat model with the aid of gene chip technology and to explore the roles of MMPs in TDVT.Methods:Totally 150 Sprague Dawley rats were randomly divided into control group (n=10) and model group (n=140). Rat models of TDVT were established by clamping the femoral vein and fixing the bilateral hind limbs. Then fixation of the hip spica with plaster bandage was conducted.According to the observation phases and/or biological situations of the femoral vein thrombosis, the model rats were further divided into 7 groups. Vascular tissues were obtained from each group through noninvasive incision into the femoral vein at corresponding time points. We adopted the Trizoi one-step method for total RNA extraction,Affymetrix RAT 230 2.0 array for detection of RNA expressions and fold change (FC) analysis for changes of differential expressions of MMPs in each group. The main outcome parameters measured included expressions of MMP-2,MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11,MMP- 12, MMP-13, MMP- 14, MMP- 16, MMP-23 and MMP-24. Gene array data of these MMPs were analyzed by the Affymetrix Microarray Analysis software (Version 5.0).Results:FC analysis showed differential expressions of MMPs in each group during the course of TDVT. At the initial period of thrombosis, MMP-2, MMP-3, MMP-7,MMP-8, MMP-9, MMP- 10, MMP-11, and MMP-24 had significantly high expression, while MMP-12, MMP-13,MMP-14, MMP-16 and MMP-23 had relatively low expression. MMPs were all highly expressed at the peak time of thrombosis. In the process of thrombus resolution,MMP-2, MMP-10, MMP-16 and MMP-24 have relatively low expression, while MMP-12, MMP-13, MMP-14,MMP-16 and MMP-23 have significantly high expression.Conclusion:MMPs may affect the process of TDVT through transcription regulation of the fibrinolysis-anti-fibrinolytic system during the course of thrombosis and thrombus resolution.

  12. Amlodipine inhibits matrix metalloproteinases expression and secretion in mouse macrophage

    Institute of Scientific and Technical Information of China (English)

    Yamin CAO; Shiwen WANG; Haiyun WU

    2005-01-01

    To investigate whether the calcium channel blocker amlodipine could inhibit macrophage matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) expression and secretion. Methods Peritoneal macrophages were isolated from BALB/C mice and incubated with low (5μg/L), middle (15μg/L) and high (305μg/L) concentrations of amlodipine, or in the medium alone (controls) for 24 hours, and the expression and secretion of MMP-2 and MM-9 of the cells were analyzed by RT-PCR and gelatin zymography. Results Compared with controls, amlodipine at low concentration had no significant effects on the expression and secretion of either MMP-2 and MMP-9 (P>0.05);at middle concentrationit it could inhibited MMP-2 and MMP-9 expressions completely and significantly reduced the secretion of MMP-9 (P<0.05); but it had no effect on the secretion of MMP-2. At high concentration it also inhibited MMP-2 and MMP-9 expression completely. Conclusion Amlodipine at 15 ìg/L inhibited the expression of MMP-2 and MMP-9 and reduced the secretion of MMP-9, suggesting that amlodipine may stabilize atherosclerotic plaque.

  13. EFFECTS OF WATER SOLUBLE TOTAL SAPONINS OF DIOSCOREA NIPPONICA ON RSC-364 CELLS SECRETING MMP-2 AND MMP-9%穿山龙水溶性总皂苷对RSC-364细胞分泌MMP-2和MMP-9的影响

    Institute of Scientific and Technical Information of China (English)

    段一娜; 杨佳琪; 王晶; 高亚贤

    2014-01-01

    目的:观察穿山龙水溶性总皂苷对大鼠滑膜成纤维细胞系RSC-364细胞分泌基质金属蛋白酶-2(MMP-2)和MMP-9的影响。方法:应用肿瘤坏死因子-α(TNF-α)和白介素17(IL-17)刺激RSC-364细胞建立类风湿性关节炎细胞模型,并以不同剂量穿山龙水溶性总皂苷进行干预,ELISA法检测细胞培养液上清MMP-2和MMP-9的水平。结果:穿山龙水溶性总皂苷各剂量组(10、20、30mg/L)在不同时间点(24、48、72h)均可明显降低TNF-α和IL-17刺激的RSC-364细胞分泌MMP-2、MMP-9水平的升高,并呈剂量依赖性(P<0.05)。结论:穿山龙水溶性总皂苷可能通过抑制RSC-364细胞分泌MMP-2和MMP-9发挥抗类风湿性关节炎的作用。%Objective: To observe the inlfuence of water soluble total saponins of Dioscorea nipponica on rat synovial ifbroblast cell line RSC-364 cells secreting matrix metalloproteinases-2 (MMP-2) and MMP-9.Methods: Interleukin-17 (IL-17) and tumor necrosis factor-α (TNF-α) were used to stimulate RSC-364 for establishing rheumatoid arthritis (RA) cell model, then the cell model was intervened by water soluble total saponins of Dioscorea nipponica in different dosage. ELISA was used to detect the MMP-2 and MMP-9 level in cultural supernatants of RSC-364 cells.Results:Water soluble total saponins of Dioscorea nipponica in different dosage at different time point could obviously reduce the level of RSC-364 cells secreting MMP-2 and MMP-9 stimulated by IL-17+ TNF-α, and showed dosage-dependent (P<0.05).Conclusions:Water soluble total saponins of Dioscorea nipponica may play a role in anti-RA by inhibiting RSC-364 cell secreting MMP-2 and MMP-9.

  14. Promoter polymorphism MMP-1 (-1607 2G/1G) and MMP-3 (-1612 5A/6A) in development of HAND and modulation of pathogenesis of HAND

    Indian Academy of Sciences (India)

    HARI OM SINGH; SHRUTI D MARATHE; SUMITRA NAIN; DHARMESH SAMANI; VIJAY NEMA; MANISHAV GHATE; R R GANGAKHEDKAR

    2017-09-01

    The pathogenesis of HIV-associated neurocognitive disorder (HAND) is modulated by host genetic susceptibility factorssuch as Matrix metalloproteinases (MMPs). Promoter polymorphism of MMP-1 and MMP-3 may modify the expression ofthe gene. Hence, we evaluated the association of MMP-1-16072G/1G and MMP-3-1612 5A/6A polymorphisms withdevelopment of HAND and the modulation of pathogenesis of HAND. We enrolled a total of 180 individuals, 50 HIVinfectedindividuals with HAND, 130 without HAND, and 150 healthy controls. Polymorphism of MMP-1 and MMP-3were genotyped by PCR-RFLP. MMP-1-1607 2G1G, -16071G/2G-1G/1G genotypes and -1607 1G allele were associatedwith the development of HAND (OR = 1.64, P = 0.05; OR = 1.45, P = 0.04; OR = 1.69, P = 0.05). MMP-1-16071G1G, MMP-3-16125A5A genotypes increased the risk for the development of HAND (OR = 1.78, P = 0.25;OR = 2.39, P = 0.13). MMP-3-1612 5A5A, -1612 6A/5A-5A/5A genotypes and -1612 5A allele were associated with thereduced risk of HAND (OR = 0.40, P = 0.05; OR = 0.53, P = 0.04; OR = 0.40, P = 0.01). Haplotype 5A1G increasedthe risk of development of HAND (OR = 1.93, P = 0.05). As observed in advanced HIV disease stage, MMP-1-16071G1G genotype enhance the risk for advancement of HIV disease (OR = 1.69, P = 0.89). MMP-3-1612 6A5A genotypeshowed higher risk for development of HAND in alcohol users (0R = 1.65, P = 0.44). MMP-1 genotype may have aninfluence on development of HAND whereas MMP3-1612 5A5A genotype may reduce risk for pathogenesis of HAND.

  15. MMP Mediated Degradation of Type VI Collagen Is Highly Associated with Liver Fibrosis - Identification and Validation of a Novel Biochemical Marker Assay

    DEFF Research Database (Denmark)

    Veidal, Sanne Skovgard; Karsdal, Morten Asser; Vassiliadis, Efstathios

    2011-01-01

    Background and Aims: During fibrogenesis, in which excessive remodeling of the extracellular matrix occurs, both the quantity of type VI collagen and levels of matrix metalloproteinases, including MMP-2 and MMP-9, increase significantly. Proteolytic degradation of type VI collagen into small...... fragments, so-called neo-epitopes, may be specific biochemical marker of liver fibrosis. The aim of this study was to develop an ELISA detecting a fragment of type VI collagen generated by MMP-2 and MMP-9, and evaluate this assay in two preclinical models of liver fibrosis. Methods: Mass spectrometric...... analysis of cleaved type VI collagen revealed a large number of protease-generated neo-epitopes. A fragment unique to type VI collagen generated by MMP-2 and MMP-9 was selected for ELISA development. The CO6-MMP assay was evaluated in two rat models of liver fibrosis: bile duct ligation (BDL) and carbon...

  16. Macrophage Metalloelastase (MMP12) Regulates Adipose Tissue Expansion, Insulin Sensitivity, and Expression of Inducible Nitric Oxide Synthase

    Science.gov (United States)

    Lee, Jung-Ting; Pamir, Nathalie; Liu, Ning-Chun; Kirk, Elizabeth A.; Averill, Michelle M.; Becker, Lev; Larson, Ilona; Hagman, Derek K.; Foster-Schubert, Karen E.; van Yserloo, Brian; Bornfeldt, Karin E.; LeBoeuf, Renee C.; Kratz, Mario

    2014-01-01

    Macrophage metalloelastase, a matrix metallopeptidase (MMP12) predominantly expressed by mature tissue macrophages, is implicated in pathological processes. However, physiological functions for MMP12 have not been described. Because mRNA levels for the enzyme increase markedly in adipose tissue of obese mice, we investigated the role of MMP12 in adipose tissue expansion and insulin resistance. In humans, MMP12 expression correlated positively and significantly with insulin resistance, TNF-α expression, and the number of CD14+CD206+ macrophages in adipose tissue. MMP12 was the most abundant matrix metallopeptidase detected by proteomic analysis of conditioned medium of M2 macrophages and dendritic cells. In contrast, it was detected only at low levels in bone marrow derived macrophages and M1 macrophages. When mice received a high-fat diet, adipose tissue mass increased and CD11b+F4/80+CD11c−macrophages accumulated to a greater extent in MMP12-deficient (Mmp12−/−) mice than in wild-type mice (Mmp12+/+). Despite being markedly more obese, fat-fed Mmp12−/− mice were more insulin sensitive than fat-fed Mmp12+/+ mice. Expression of inducible nitric oxide synthase (Nos2) by Mmp12−/− macrophages was significantly impaired both in vivo and in vitro, suggesting that MMP12 might mediate nitric oxide production during inflammation. We propose that MMP12 acts as a double-edged sword by promoting insulin resistance while combatting adipose tissue expansion. PMID:24914938

  17. Green tea polyphenol epigallocatechin-3-gallate suppresses rat hepatic stellate cell invasion by inhibition of MMP-2 expression and its activation

    Institute of Scientific and Technical Information of China (English)

    Mao-chuan ZHEN; Xiao-hui HUANG; Qian WANG; Kai SUN; Yun-jian LIU; Wen LI; Long-Juan ZHANG; Liang-qi CAO; Xi-ling CHEN

    2006-01-01

    Aim: Epigallocatechin-3-gallate (EGCG) is the major component of green tea polyphenols, whose wide range of biological properties includes anti-fibrogenic activity. Matrix metalloproteinases (MMP) that participate in extracellular matrix degradation are involved in the development of hepatic fibrosis. The present study investigates whether EGCG inhibits activation of the major gelatinase matrix metalloproteinase-2 (MMP-2) in rat hepatic stellate cells (HSC). Methods: The expression of MMP-2, tissue inhibitors of metalloproteinases-2 (TIMP-2), and membrane-type 1-MMP (MT1-MMP) was assessed by RT-PCR and Western blot analyses. MMP-2 activity was evaluated by zymography and MT1-MMP activity was assessed by an enzymatic assay. HSC migration was measured by a wound healing assay and cell invasion was performed using Transwell cell culture chambers. Results: The expression of MMP-2 mRNA and protein in HSC was substantially reduced by EGCG treatment. EGCG treatment also reduced con-canavalin A (ConA)-induced activation of secreted MMP-2 and reduced MT1-MMP activity in a dose-dependent manner. In addition, EGCG inhibited either HSC migration or invasion. Conclusion: The abilities of EGCG to suppress MMP-2 activation and HSC invasiveness suggest that EGCG may be useful in the treatment and prevention of hepatic fibrosis.

  18. Immunohistochemical expression of MMP-14 and MMP-2, and MMP-2 activity during human ovarian follicular development

    NARCIS (Netherlands)

    Vos, M.C.; Wurff, A.A. van der; Last, J.T.; Boed, E.A. de; Smeenk, J.M.J.; Kuppevelt, T.H. van; Massuger, L.F.A.G.

    2014-01-01

    BACKGROUND: The aim of this study was to investigate the presence of MMP-14 and MMP-2 during human ovarian follicular development using immunohistochemistry, and the activity of MMP-2 in follicular fluid using zymography. METHODS: Ovarian tissue collected from the archives of the Department of Patho

  19. Increased expression of MMP-9 and IL-8 are correlated with poor prognosis of Bladder Cancer

    Directory of Open Access Journals (Sweden)

    Reis Sabrina

    2012-06-01

    Full Text Available Abstract Background Extracellular matrix homeostasis is strictly maintained by a coordinated balance between the expression of metalloproteinases (MMPs and their inhibitors. The purpose of this study was to investigate whether the expression of MMP-9, MMP-2 and its specific inhibitors, are expressed in a reproducible, specific pattern and if the profiles are related to prognosis in Bladder Cancer (BC. Methods MMP-9, MMP-2 and its specific inhibitors expression levels were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR in fresh-frozen malignant tissue collected from 40 patients with BC submitted to transurethral resection of bladder. The control group consisted of normal bladder tissue from five patients who had undergone retropubic prostatectomy to treat benign prostatic hyperplasia. Results MMP-9 was overexpressed in 59.0 % of patients, and MMP-2, TIMP-1, TIMP-2, MMP-14, RECK and IL-8 was underexpressed in most of the patients. Regarding prognostic parameters we observed that high-grade tumors exhibited significantly higher levels of MMP-9 and IL-8 (p = 0.012, p = 0.003. Invasive tumors (pT1-pT2 had higher expression levels of MMP-9 than superficial tumors (pTa (p = 0.026. The same was noted for IL-8 that was more expressed by invasive tumors (p = 0.015, p = 0.048. Most importantly tumor recurrence was related with higher levels of both MMP-9 (p = 0.003 and IL-8 (p = 0.005. Conclusion We have demonstrated that the overexpression of MMP-9 and higher expression of IL-8 are related to unfavorable prognostic factors of urothelial bladder cancer and tumor recurrence and may be useful in the follow up of the patients.

  20. LPS responsiveness and neutrophil chemotaxis in vivo require PMN MMP-8 activity.

    Directory of Open Access Journals (Sweden)

    Angus M Tester

    Full Text Available We identify matrix metalloproteinase (MMP-8, the polymorphonuclear (PMN leukocyte collagenase, as a critical mediator initiating lipopolysaccharide (LPS-responsiveness in vivo. PMN infiltration towards LPS is abrogated in Mmp8-null mice. MMP-8 cleaves LPS-induced CXC chemokine (LIX at Ser(4-Val(5 and Lys(79-Arg(80. LIX bioactivity is increased upon N-terminal cleavage, enhancing intracellular calcium mobilization and chemotaxis upon binding its cognate receptor, CXCR2. As there is no difference in PMN chemotaxis in Mmp8-null mice compared with wild-type mice towards synthetic analogues of MMP-8-cleaved LIX, MMP-8 is not essential for extravasation or cell migration in collagenous matrices in vivo. However, with biochemical redundancy between MMPs 1, 2, 9, and 13, which also cleave LIX at position 4 approximately 5, it was surprising to observe such a markedly reduced PMN infiltration towards LPS and LIX in Mmp8-/- mice. This lack of physiological redundancy in vivo identifies MMP-8 as a key mediator in the regulation of innate immunity. Comparable results were found with CXCL8/IL-8 and CXCL5/ENA-78, the human orthologues of LIX. MMP-8 cleaves CXCL8 at Arg(5-Ser(6 and at Val(7-Leu(8 in CXCL5 to activate respective chemokines. Hence, rather than collagen, these PMN chemoattractants are important MMP-8 substrates in vivo; PMN-derived MMP-8 cleaves and activates LIX to execute an in cis PMN-controlled feed-forward mechanism to orchestrate the initial inflammatory response and promote LPS responsiveness in tissue.

  1. Expression of MMP-2, MMP-9 and its tissue inhibitors in patients with endometriosis%子宫内膜异位症患者MMP-2、 MMP-9及其组织抑制因子的表达

    Institute of Scientific and Technical Information of China (English)

    潘孝勇; 刘倩如; 郭美丽; 曾松芳

    2016-01-01

    目的 探讨血清和腹腔液中基质金属蛋白酶MP-2、MMP-9及其组织抑制因子TIMP-1、TIMP-2水平与子宫内膜异位症(EMs)发病的关系.方法 收集2014年1月~2015年12月确诊的83例EMs患者和35例对照组血清和腹腔液,用酶联免疫吸附法(ELISA)检测MMP-2、MMP-9、TIMP-1和TIMP-2的浓度.结果 EMs组血清和腹水中MMP-2 和MMP-9度显著高于对照组,TIMP-1和TIMP-2显著低于对照组(P<0.05);Ⅲ-Ⅳ期患者组MMP-2和MMP-9水平显著高于Ⅰ-Ⅱ期组,TIMP-1和TIMP-2水平显著低于Ⅰ-Ⅱ期组(P<0.05).结论 EMs患者MMP-2和MMP-9高表达,TIMP-1和TIMP-2低表达,MMP-2/TIMP-2和MMP-9/TIMP-1的比值增高,使异位内膜组织具有更强的侵袭力,可能在子宫内膜异位症的发生发展中起重要作用.%Objective:To explore the expression and significance of matrix metallopmteinase-2,-9 (MMP-2,MMP-9) and tissue inhibitor metalloproteinase-1,-2 (TIMP-1,TIMP-2) in endometfiosis (EMs) in serum and peritonoeal fluid.Methods:The serum and peritoneal fluid was obtained from 83 cases EMs patients and 35 cases control group from Jan.2014 to Dec.2015.The concentration of MMP-2,MMP-9,TIMP-1 and TIMP-2 was detected by enzyme-linked immunosorbent assay (ELISA) method.Results:The concentration of MMP-2 and MMP-9 in Serum and peritoneal fluid of EMs group is significantly higher than that of the control group,the concentration of TIMP-1 and TIMP-2 in EMs group was significantly lower than that of the control group (P<0.05).The concentration ofMMP-2 and MMP-9 in]Ⅲ-Ⅳ stage of EMs group is significantly higher than that of Ⅰ-Ⅱ stage,the concentration of TIMP-1 and TIMP-2 in]Ⅲ-Ⅳ stage of EMs group is significantly lower than that of Ⅰ-Ⅱ stage (P<0.05).Conclusion:The increased expression of MMP-2,MMP-9 and the decreased expression of TIMP-1,TIMP-2 in EMs patients result the higher ratio of MMP-2/TIMP-2 and MMP-9/TIMP-1.It can make ectopic endometrial tissues have a greater capactity to

  2. Multiple essential MT1-MMP functions in tooth root formation, dentinogenesis, and tooth eruption.

    Science.gov (United States)

    Xu, H; Snider, T N; Wimer, H F; Yamada, S S; Yang, T; Holmbeck, K; Foster, B L

    2016-01-01

    Membrane-type matrix metalloproteinase 1 (MT1-MMP) is a transmembrane zinc-endopeptidase that breaks down extracellular matrix components, including several collagens, during tissue development and physiological remodeling. MT1-MMP-deficient mice (MT1-MMP(-/-)) feature severe defects in connective tissues, such as impaired growth, osteopenia, fibrosis, and conspicuous loss of molar tooth eruption and root formation. In order to define the functions of MT1-MMP during root formation and tooth eruption, we analyzed the development of teeth and surrounding tissues in the absence of MT1-MMP. In situ hybridization showed that MT1-MMP was widely expressed in cells associated with teeth and surrounding connective tissues during development. Multiple defects in dentoalveolar tissues were associated with loss of MT1-MMP. Root formation was inhibited by defective structure and function of Hertwig's epithelial root sheath (HERS). However, no defect was found in creation of the eruption pathway, suggesting that tooth eruption was hampered by lack of alveolar bone modeling/remodeling coincident with reduced periodontal ligament (PDL) formation and integration with the alveolar bone. Additionally, we identified a significant defect in dentin formation and mineralization associated with the loss of MT1-MMP. To segregate these multiple defects and trace their cellular origin, conditional ablation of MT1-MMP was performed in epithelia and mesenchyme. Mice featuring selective loss of MT1-MMP activity in the epithelium were indistinguishable from wild type mice, and importantly, featured a normal HERS structure and molar eruption. In contrast, selective knock-out of MT1-MMP in Osterix-expressing mesenchymal cells, including osteoblasts and odontoblasts, recapitulated major defects from the global knock-out including altered HERS structure, short roots, defective dentin formation and mineralization, and reduced alveolar bone formation, although molars were able to erupt. These data

  3. MMP20 Promotes a Smooth Enamel Surface, a Strong DEJ, and a Decussating Enamel Rod Pattern

    Science.gov (United States)

    Bartlett, John D.; Skobe, Ziedonis; Nanci, Antonio; Smith, Charles E.

    2012-01-01

    Mutations of the Matrix metalloproteinase-20 (MMP20, enamelysin) gene cause autosomal recessive amelogenesis imperfecta and Mmp20 ablated mice also have malformed dental enamel. Here we show that Mmp20 null mouse secretory stage ameloblasts maintained a columnar shape and were present as a single layer of cells. However, the null maturation stage ameloblasts covered extraneous nodules of ectopic calcified material formed at the enamel surface. Remarkably, nodule formation occurs in null mouse enamel when MMP20 is normally no longer expressed. The malformed enamel in Mmp20 null teeth was loosely attached to the dentin and the entire enamel layer tended to separate from the dentin indicative of a faulty DEJ. The enamel rod pattern was also altered in Mmp20 null mice. Each enamel rod is formed by a single ameloblast and is a mineralized record of the migration path of the ameloblast that formed it. The Mmp20 null mouse enamel rods were grossly malformed or were absent indicating that the ameloblasts do not migrate properly when backing away from the DEJ. Thus, MMP20 is required for ameloblast cell movement necessary to form the decussating enamel rod patterns, for the prevention of ectopic mineral formation, and to maintain a functional DEJ. PMID:22243247

  4. MMP-3 Deficiency Alleviates Endotoxin-Induced Acute Inflammation in the Posterior Eye Segment

    Directory of Open Access Journals (Sweden)

    Inge Van Hove

    2016-11-01

    Full Text Available Matrix metalloproteinase-3 (MMP-3 is known to mediate neuroinflammatory processes by activating microglia, disrupting blood–central nervous system barriers and supporting neutrophil influx into the brain. In addition, the posterior part of the eye, more specifically the retina, the retinal pigment epithelium (RPE and the blood–retinal barrier, is affected upon neuroinflammation, but a role for MMP-3 during ocular inflammation remains elusive. We investigated whether MMP-3 contributes to acute inflammation in the eye using the endotoxin-induced uveitis (EIU model. Systemic administration of lipopolysaccharide induced an increase in MMP-3 mRNA and protein expression level in the posterior part of the eye. MMP-3 deficiency or knockdown suppressed retinal leukocyte adhesion and leukocyte infiltration into the vitreous cavity in mice subjected to EIU. Moreover, retinal and RPE mRNA levels of intercellular adhesion molecule 1 (Icam1, interleukin 6 (Il6, cytokine-inducible nitrogen oxide synthase (Nos2 and tumor necrosis factor α (Tnfα, which are key molecules involved in EIU, were clearly reduced in MMP-3 deficient mice. In addition, loss of MMP-3 repressed the upregulation of the chemokines monocyte chemoattractant protein (MCP-1 and (C-X-C motif ligand 1 (CXCL1. These findings suggest a contribution of MMP-3 during EIU, and its potential use as a therapeutic drug target in reducing ocular inflammation.

  5. Serum and Saliva MMP-3 in Patients with OLP and Oral SCC.

    Science.gov (United States)

    Agha-Hosseini, Farzaneh; Mirzaii-Dizgah, Iraj; Mahboobi, Nima; Shirazian, Shiva; Harirchi, Iraj

    2015-02-01

    Matrix metalloproteinase-3 (MMP-3) plays a key role in development of cancer. The purpose of this study was to assess MMP-3 in the serum and saliva of patients with oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). Thirty patients with OLP (8 reticular and 22 erosive forms), and 20 patients with OSCC (6 in low stage and 14 in advanced stage), were enrolled in this study, conducted at the Cancer Department, Clinic of Oral Medicine, Tehran University of Medical Sciences. The serum and saliva MMP-3 was assayed by ELISA method. Statistical analysis of the Student's t-test, ANOVA and Pearson correlation coefficient was performed. The mean saliva and serum levels of MMP-3 were significantly higher in patients with OSCC compared with OLP. The serum and saliva MMP-3 concentrations increased from reticular form of OLP to erosive form of OLP, and increased further to low stage of OSCC and advanced stage of OSCC. Serum MMP-3 correlated significantly with unstimulated (r = 0.310, p = 0.038) and stimulated (r = 0.365, p saliva MMP-3. Serum and saliva MMP-3 levels appear associated with OLP and OSCC.

  6. MT1-MMP-mediated basement membrane remodeling modulates renal development

    Energy Technology Data Exchange (ETDEWEB)

    Riggins, Karen S.; Mernaugh, Glenda [Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Su, Yan; Quaranta, Vito [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Koshikawa, Naohiko; Seiki, Motoharu [Division of Cancer Cell Research, Institute of Medical Science, The University of Tokyo, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Pozzi, Ambra [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Research Medicine, Veterans Affairs Hospital, Nashville, TN 37232 (United States); Zent, Roy, E-mail: roy.zent@vanderbilt.edu [Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Department of Research Medicine, Veterans Affairs Hospital, Nashville, TN 37232 (United States)

    2010-10-15

    Extracellular matrix (ECM) remodeling regulates multiple cellular functions required for normal development and tissue repair. Matrix metalloproteinases (MMPs) are key mediators of this process and membrane targeted MMPs (MT-MMPs) in particular have been shown to be important in normal development of specific organs. In this study we investigated the role of MT1-MMP in kidney development. We demonstrate that loss of MT1-MMP leads to a renal phenotype characterized by a moderate decrease in ureteric bud branching morphogenesis and a severe proliferation defect. The kidneys of MT1-MMP-null mice have increased deposition of collagen IV, laminins, perlecan, and nidogen and the phenotype is independent of the MT-1MMP target, MMP-2. Utilizing in vitro systems we demonstrated that MTI-MMP proteolytic activity is required for renal tubule cells to proliferate in three dimensional matrices and to migrate on collagen IV and laminins. Together these data suggest an important role for MT1-MMP in kidney development, which is mediated by its ability to regulate cell proliferation and migration by proteolytically cleaving kidney basement membrane components.

  7. High-concentration glucose enhances invasion in invasive ductal breast carcinoma by promoting Glut1/MMP2/MMP9 axis expression.

    Science.gov (United States)

    Sun, Xian-Fu; Shao, Ying-Bo; Liu, Ming-Ge; Chen, Qi; Liu, Zhao-Jun; Xu, Bin; Luo, Su-Xia; Liu, Hui

    2017-05-01

    Type 2 diabetes mellitus (T2DM) has been considered to be a risk factor for numerous human cancers. Hyperglycemia is one of the most direct internal environmental changes for patients with T2DM. Increasing evidence reveals that a high concentration of glucose can promote tumor progression, while its role for migration and invasion of invasive ductal breast carcinoma (IDBC) cells remains unclear. In the present study, it was demonstrated that IDBC patients with T2DM suffered an increased tumor size and more frequent lymphatic and distant metastasis compared with those without T2DM (P<0.05). MCF-7 breast carcinoma cells, which were cultured in a high glucose concentration medium (25.00 mM), exhibited increased invasion (P<0.05). In addition, the expression of glucose transporters (Gluts), matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9) in IDBC tissues with T2DM was significantly higher compared to those without T2DM. Downregulation of glucose transporter 1 (Glut1) by small interfering RNA may markedly suppress MCF-7 cell invasion as well as the expression of MMP2 and MMP9. These results suggest that T2DM can affect the malignant features of tumors in IDBC. The high glucose concentration in the tumor microenvironment may enhance IDBC invasion via upregulating Glut1/MMP2/MMP9 axis expression.

  8. Epb41l3 suppresses esophageal squamous cell carcinoma invasion and inhibits MMP2 and MMP9 expression.

    Science.gov (United States)

    Zeng, Rong; Huang, Jun-Peng; Li, Xu Feng; Xiong, Wei-Bin; Wu, Gang; Jiang, Zhao-Jing; Song, Shu-Jie; Li, Ji-Qiang; Zheng, Yan-Fang; Zhang, Ji-Ren

    2016-04-01

    EPB41L3 may play a role as a metastasis suppressor by supporting regular arrangements of actin stress fibres and alleviating the increase in cell motility associated with enhanced metastatic potential. Downregulation of epb41l3 has been observed in many cancers, but the role of this gene in esophageal squamous cell carcinoma (ESCC) remains unclear. Our study aimed to determine the effect of epb41l3 on ESCC cell migration and invasion. We investigated epb41l3 protein expression in tumour and non-tumour tissues by immunohistochemical staining. Expression in the non-neoplastic human esophageal cell line Het-1a and four ESCC cell lines - Kyse150, Kyse510, Kyse450 and Caes17 - was assessed by quantitative Polymerase Chain Reaction (qPCR) and Western blotting. Furthermore, an EPB41L3 overexpression plasmid and EPB41L3-specific small interfering RNA were used to upregulate EPB41L3 expression in Kyse150 cells and to downregulate EPB41L3 expression in Kyse450 cells, respectively. Cell migration and invasion were evaluated by wound healing and transwell assays, respectively. The expression levels of p-AKT, matrix metalloproteinase (MMP)2 and MMP9 were evaluated. Expression of epb41l3 was significantly lower in tumour tissues than in non-tumour tissues and in ESCC cell lines compared with the Het-1a cell line. Kyse450 and Caes17 cells exhibited higher expression of epb41l3 than Kyse150 and Kyse510 cells. Overexpressing epb41l3 decreased Kyse150 cell migration and invasion, whereas EPB41L3-specific small interfering RNA silencing increased these functions in Kyse450 cells. Furthermore, overexpressing epb41l3 led to downregulation of MMP2 and MMP9 in Kyse150 and Kyse510 cells. Our findings reveal that EPB41L3 suppresses tumour cell invasion and inhibits MMP2 and MMP9 expression in ESCC cells.

  9. 2-Deoxy glucose regulate MMP-9 in a SIRT-1 dependent and NFkB independent mechanism.

    Science.gov (United States)

    Edatt, Lincy; Haritha, K; Sruthi, T V; Aswini, P; Sameer Kumar, V B

    2016-12-01

    MMP9 is a member of the family of zinc-containing endopeptidases which degrade various components of the extracellular matrix, thereby regulating matrix remodeling. Since matrix remodeling plays an important role during growth and progression of cancer and considering the fact that, tumor cells switch to aerobic glycolysis as its major energy source, this study was designed to analyze if partial inhibition of glycolysis (the major energy pathway during hypoxia) can be used as a means to control matrix remodeling in terms of MMP9 activity and expression. For this, human epithelial carcinoma cells were treated with glycolytic inhibitor, 2-deoxy glucose (2DG) at sub-lethal concentrations followed by analysis of the expression and activity of MMP2 and MMP9. The experimental findings demonstrate that exposure of cancer cells to glycolytic inhibitor at concentration that does not induce ER stress, downregulates the activity and expression of MMP9 without affecting the expression levels and activity of MMP2. Further mechanistic analysis revealed that the regulation of MMP9 was mediated in a SIRT-1 dependent mechanism and did not alter the NFkB signaling pathway. The overall results presented here, therefore suggest that the use of glycolytic inhibitor, 2DG at concentration that do not affect cell viability or induce ER stress can be an effective strategy to control matrix remodeling.

  10. Prospective study of MMP7 serum levels in the diagnosis of cholangiocarcinoma

    Institute of Scientific and Technical Information of China (English)

    Kawin; Leelawat; Siriluck; Narong; Jerasak; Wannaprasert; Thawee; Ratanashu-ek

    2010-01-01

    AIM: To determine whether the serum level of matrix metalloproteinase-7 (MMP7) has the potential to diagnosis cholangiocarcinoma from benign biliary tract diseases. METHODS: This study was performed according to the PRoBE (a prospective-specimen-collection, retrospectiveblinded-evaluation) design. A total of 187 patients with obstructive jaundice were consecutively enrolled. After the diagnostic status of these patients was ascertained, their levels of serum MMP7 were assayed and compared with serum carbohy...

  11. Measurement of MMP-9 and -12 degraded elastin (ELM) provides unique information on lung tissue degradation

    DEFF Research Database (Denmark)

    Skjøt-Arkil, Helene; Clausen, Rikke E; Nguyen, Quoc Hai Trieu;

    2012-01-01

    Elastin is an essential component of selected connective tissues that provides a unique physiological elasticity. Elastin may be considered a signature protein of lungs where matrix metalloprotease (MMP) -9-and -12, may be considered the signature proteases of the macrophages, which in part...... are responsible for tissue damage during disease progression. Thus, we hypothesized that a MMP-9/-12 generated fragment of elastin may be a relevant biochemical maker for lung diseases....

  12. Matrix metalloproteinases as therapeutic targets for idiopathic pulmonary fibrosis.

    Science.gov (United States)

    Craig, Vanessa J; Zhang, Li; Hagood, James S; Owen, Caroline A

    2015-11-01

    Idiopathic pulmonary fibrosis (IPF) is a restrictive lung disease that is associated with high morbidity and mortality. Current medical therapies are not fully effective at limiting mortality in patients with IPF, and new therapies are urgently needed. Matrix metalloproteinases (MMPs) are proteinases that, together, can degrade all components of the extracellular matrix and numerous nonmatrix proteins. MMPs and their inhibitors, tissue inhibitors of MMPs (TIMPs), have been implicated in the pathogenesis of IPF based upon the results of clinical studies reporting elevated levels of MMPs (including MMP-1, MMP-7, MMP-8, and MMP-9) in IPF blood and/or lung samples. Surprisingly, studies of gene-targeted mice in murine models of pulmonary fibrosis (PF) have demonstrated that most MMPs promote (rather than inhibit) the development of PF and have identified diverse mechanisms involved. These mechanisms include MMPs: (1) promoting epithelial-to-mesenchymal transition (MMP-3 and MMP-7); (2) increasing lung levels or activity of profibrotic mediators or reducing lung levels of antifibrotic mediators (MMP-3, MMP-7, and MMP-8); (3) promoting abnormal epithelial cell migration and other aberrant repair processes (MMP-3 and MMP-9); (4) inducing the switching of lung macrophage phenotypes from M1 to M2 types (MMP-10 and MMP-28); and (5) promoting fibrocyte migration (MMP-8). Two MMPs, MMP-13 and MMP-19, have antifibrotic activities in murine models of PF, and two MMPs, MMP-1 and MMP-10, have the potential to limit fibrotic responses to injury. Herein, we review what is known about the contributions of MMPs and TIMPs to the pathogenesis of IPF and discuss their potential as therapeutic targets for IPF.

  13. Relaxin stimulates MMP-2 and α-smooth muscle actin expression by human periodontal ligament cells

    NARCIS (Netherlands)

    Henneman, S.; Bildt, M.M.; Groot, J. de; Kuijpers-Jagtman, A.M.; Von den Hoff, J.W.

    2008-01-01

    The main cells in the periodontal ligament (PDL) are the fibroblasts, which play an important role in periodontal remodelling. Matrix metalloproteinases (MMPs) are largely responsible for the degradation of extracellular matrix proteins in the PDL. Previous studies have indicated that MMP production

  14. Relaxin stimulates MMP-2 and alpha-smooth muscle actin expression by human periodontal ligament cells.

    NARCIS (Netherlands)

    Henneman, S.; Bildt, M.M.; Degroot, J.; Kuijpers-Jagtman, A.M.; Hoff, J.W. Von den

    2008-01-01

    The main cells in the periodontal ligament (PDL) are the fibroblasts, which play an important role in periodontal remodelling. Matrix metalloproteinases (MMPs) are largely responsible for the degradation of extracellular matrix proteins in the PDL. Previous studies have indicated that MMP production

  15. Human MMP28 expression is unresponsive to inflammatory stimuli and does not correlate to the grade of intervertebral disc degeneration

    Directory of Open Access Journals (Sweden)

    Klawitter Marina

    2011-07-01

    Full Text Available Abstract Background MMP28 (epilysin is a recently discovered member of the MMP (matrix metalloproteinase family that is, amongst others, expressed in osteoarthritic cartilage and intervertebral disc (IVD tissue. In this study the hypothesis that increased expression of MMP28 correlates with higher grades of degeneration and is stimulated by the presence of proinflammatory molecules was tested. Gene expression levels of MMP28 were investigated in traumatic and degenerative human IVD tissue and correlated to the type of disease and the degree of degeneration (Thompson grade. Quantification of MMP28 gene expression in human IVD tissue or in isolated cells after stimulation with the inflammatory mediators lipopolysaccharide (LPS, interleukin (IL-1β, tumor necrosis factor (TNF-α or the histondeacetylase inhibitor trichostatin A was performed by real-time RT PCR. Results While MMP28 expression was increased in individual cases with trauma or disc degeneration, there was no significant correlation between the grade of disease and MMP28 expression. Stimulation with LPS, IL-1β, TNF-α or trichostatin A did not alter MMP28 gene expression at any investigated time point or any concentration. Conclusions Our results demonstrate that gene expression of MMP28 in the IVD is not regulated by inflammatory mechanisms, is donor-dependent and cannot be positively or negatively linked to the grade of degeneration and only weakly to the occurrence of trauma. New hypotheses and future studies are needed to find the role of MMP28 in the intervertebral disc.

  16. Crosstalk between obesity and MMP-9 in cardiac remodelling -a cross-sectional study in apparent treatment-resistant hypertension.

    Science.gov (United States)

    Ritter, Alessandra Mileni Versuti; de Faria, Ana Paula; Barbaro, Natália; Sabbatini, Andréa Rodrigues; Corrêa, Nathália Batista; Brunelli, Veridiana; Amorim, Rivadavio; Modolo, Rodrigo; Moreno, Heitor

    2017-04-01

    The balance between matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) plays a key role in the development of hypertension and obesity. We aimed to evaluate the levels of MMP-2 and 9 and TIMP-2 and -1 in obese and non-obese apparent treatment-resistant hypertensive subjects (aTRH) and its association with cardiac hypertrophy. This cross-sectional study enrolled 122 subjects and divided into obese aTRH (n = 67) and non-obese (n = 55) group. Clinical and biochemical data were compared between both groups, including office BP, ambulatory BP, plasma MMP-2 and 9, TIMP-2 and 1 and left ventricular mass index (LVMI). We found higher MMP-9 levels and MMP-9/TIMP-1 ratio in obese aTRH subjects but no difference in MMP-2 and TIMP-1 levels. Obesity influenced MMP-9 levels [β = 20.8 SE =8.6, p = 0.02) independently of potential confounders. In addition, we found a positive correlation between MMP-9 and anthropomorphic parameters. Finally, obese aTRH subjects with left ventricular hypertrophy (LVH) had greater MMP-9 levels compared with non-obese with LVH. Our study suggests that MMP-9 levels are influenced by obesity and may directly participate in the progressive LV remodelling process, suggesting a possible role for a higher cardiovascular risk in apparent resistant hypertensive subjects.

  17. MMP1-1607 polymorphism increases the risk for periapical lesion development through the upregulation MMP-1 expression in association with pro-inflammatory milieu elements

    Science.gov (United States)

    TROMBONE, Ana Paula Favaro; CAVALLA, Franco; SILVEIRA, Elcia Maria Varize; ANDREO, Camile Bermejo; FRANCISCONI, Carolina Favaro; FONSECA, Angélica Cristina; LETRA, Ariadne; SILVA, Renato Menezes; GARLET, Gustavo Pompermaier

    2016-01-01

    ABSTRACT Increased matrix metalloproteinases (MMPs) activity is a hallmark of periapical granulomas. However, the factors underlying the MMPs expression modulation in healthy and diseased periapical tissues remains to be determined. Objective In this study, we evaluated the association between the MMP1-1607 polymorphism (rs1799750) and pro-inflammatory milieu elements with MMP-1 mRNA levels in vivo. Material and Methods MMP1-1607 SNP and the mRNA levels of MMP-1, TNF-a, IFN-g, IL-17A, IL-21, IL-10, IL-4, IL-9, and FOXp3 were determined via RealTimePCR in DNA/RNA samples from patients presenting periapical granulomas (N=111, for both genotyping and expression analysis) and control subjects (N=214 for genotyping and N=26 for expression analysis). The Shapiro-Wilk, Fisher, Pearson, Chi-square ordinal least squares regression tests were used for data analysis (p<0.05 was considered statistically significant). Results The MMP1-1607 1G/2G and 1G/2G+2G/2G genotypes were significantly more prevalent in the patients than in controls, comprising a risk factor for periapical lesions development. MMP-1 mRNA levels were higher in periapical lesions than in healthy periodontal ligament samples, as well as higher in active than in inactive lesions. The polymorphic allele 2G carriers presented a significantly higher MMP-1 mRNA expression when compared with the 1G/1G genotype group. The ordered logistic regression demonstrated a significant correlation between the genetic polymorphism and the expression levels of MMP-1. Additionally, the pro- and anti-inflammatory cytokines IL-17A, IFN-g, TNF-a, IL-21, IL-10, IL-9, and IL-4 were significant as complementary explanatory variables of MMP-1 expression. Conclusion The MMP1-1607 SNP was identified as a risk factor for periapical lesions development, possibly due to its association with increased MMP-1 mRNA levels in periapical lesions. The MMP-1 expression is also under the control of the inflammatory milieu elements, being the

  18. Mycobacterium tuberculosis Upregulates TNF-α Expression via TLR2/ERK Signaling and Induces MMP-1 and MMP-9 Production in Human Pleural Mesothelial Cells.

    Directory of Open Access Journals (Sweden)

    Wei-Lin Chen

    Full Text Available Tumor necrosis factor (TNF-α and matrix metalloproteinases (MMPs are elevated in pleural fluids of tuberculous pleuritis (TBP where pleural mesothelial cells (PMCs conduct the first-line defense against Mycobacterium tuberculosis (MTB. However, the clinical implication of TNF-α and MMPs in TBP and the response of PMCs to MTB infection remain unclear.We measured pleural fluid levels of TNF-α and MMPs in patients with TBP (n = 18 or heart failure (n = 18 as controls. Radiological scores for initial effusion amount and residual pleural fibrosis at 6-month follow-up were assessed. In vitro human PMC experiments were performed to assess the effect of heat-killed M. tuberculosis H37Ra (MTBRa on the expression of TNF-α and MMPs.As compared with controls, the effusion levels of TNF-α, MMP-1 and MMP-9 were significantly higher and correlated positively with initial effusion amount in patients with TBP, while TNF-α and MMP-1, but not MMP-9, were positively associated with residual pleural fibrosis of TBP. Moreover, effusion levels of TNF-α had positive correlation with those of MMP-1 and MMP-9 in TBP. In cultured PMCs, MTBRa enhanced TLR2 and TLR4 expression, activated ERK signaling, and upregulated TNF-α mRNA and protein expression. Furthermore, knockdown of TLR2, but not TLR4, significantly inhibited ERK phosphorylation and TNF-α expression. Additionally, both MTBRa and TNF-α markedly induced MMP-1 and MMP-9 synthesis in human PMCs, and TNF-α neutralization substantially reduced the production of MMP-1, but not MMP-9, in response to MTBRa stimulation.MTBRa activates TLR2/ERK signalings to induce TNF-α and elicit MMP-1 and MMP-9 in human PMCs, which are associated with effusion volume and pleural fibrosis and may contribute to pathogenesis of TBP. Further investigation of manipulation of TNF-α and MMP expression in pleural mesothelium may provide new insights into the mechanisms and rational treatment strategies for TBP.

  19. MT1-MMP-dependent invasion is regulated by TI-VAMP/VAMP7.

    Science.gov (United States)

    Steffen, Anika; Le Dez, Gaëlle; Poincloux, Renaud; Recchi, Chiara; Nassoy, Pierre; Rottner, Klemens; Galli, Thierry; Chavrier, Philippe

    2008-06-24

    Proteolytic degradation of the extracellular matrix (ECM) is one intrinsic property of metastatic tumor cells to breach tissue barriers and to disseminate into different tissues. This process is initiated by the formation of invadopodia, which are actin-driven, finger-like membrane protrusions. Yet, little is known on how invadopodia are endowed with the functional machinery of proteolytic enzymes [1, 2]. The key protease MT1-MMP (membrane type 1-matrix metalloproteinase) confers proteolytic activity to invadopodia and thus invasion capacity of cancer cells [3-6]. Here, we report that MT1-MMP-dependent matrix degradation at invadopodia is regulated by the v-SNARE TI-VAMP/VAMP7, hence providing the molecular inventory mediating focal degradative activity of cancer cells. As observed by TIRF microscopy, MT1-MMP-mCherry and GFP-VAMP7 were simultaneously detected at proteolytic sites. Functional ablation of VAMP7 decreased the ability of breast cancer cells to degrade and invade in a MT1-MMP-dependent fashion. Moreover, the number of invadopodia was dramatically decreased in VAMP7- and MT1-MMP-depleted cells, indicative of a positive-feedback loop in which the protease as a cargo of VAMP7-targeted transport vesicles regulates maturation of invadopodia. Collectively, these data point to a specific role of VAMP7 in delivering MT1-MMP to sites of degradation, maintaining the functional machinery required for invasion.

  20. Evolutionary Analysis Predicts Sensitive Positions of MMP20 and Validates Newly- and Previously-Identified MMP20 Mutations Causing Amelogenesis Imperfecta

    Directory of Open Access Journals (Sweden)

    Barbara Gasse

    2017-06-01

    Full Text Available Amelogenesis imperfecta (AI designates a group of genetic diseases characterized by a large range of enamel disorders causing important social and health problems. These defects can result from mutations in enamel matrix proteins or protease encoding genes. A range of mutations in the enamel cleavage enzyme matrix metalloproteinase-20 gene (MMP20 produce enamel defects of varying severity. To address how various alterations produce a range of AI phenotypes, we performed a targeted analysis to find MMP20 mutations in French patients diagnosed with non-syndromic AI. Genomic DNA was isolated from saliva and MMP20 exons and exon-intron boundaries sequenced. We identified several homozygous or heterozygous mutations, putatively involved in the AI phenotypes. To validate missense mutations and predict sensitive positions in the MMP20 sequence, we evolutionarily compared 75 sequences extracted from the public databases using the Datamonkey webserver. These sequences were representative of mammalian lineages, covering more than 150 million years of evolution. This analysis allowed us to find 324 sensitive positions (out of the 483 MMP20 residues, pinpoint functionally important domains, and build an evolutionary chart of important conserved MMP20 regions. This is an efficient tool to identify new- and previously-identified mutations. We thus identified six functional MMP20 mutations in unrelated families, finding two novel mutated sites. The genotypes and phenotypes of these six mutations are described and compared. To date, 13 MMP20 mutations causing AI have been reported, making these genotypes and associated hypomature enamel phenotypes the most frequent in AI.

  1. The effect of gallium nitrate on synoviocyte MMP activity.

    Science.gov (United States)

    Panagakos, F S; Kumar, E; Venescar, C; Guidon, P

    2000-02-01

    Gallium, a group IIIa metal salt, has been demonstrated to be an effective immunosuppressive agent. Gallium has also been shown to inhibit the production of inflammatory cytokines, such as IL-1beta, produced by macrophage-like cells in vitro. To further characterize the effects of gallium on the inflammatory process, we examined the effects of gallium nitrate on matrix metalloproteinase (MMP) activity utilizing the rabbit synoviocyte cell line HIG-82. HIG-82 cells were incubated with IL-1beta and TPA, with and without increasing concentrations of gallium nitrate. Conditioned medium was collected and assayed for MMP activity using a synthetic substrate and substrate gel zymography. IL-1beta and TPA alone induced MMP activity in HIG-82 cells. A dose-dependent inhibition of IL-1beta and TPA stimulated MMP activity by gallium nitrate at increasing concentrations was observed. This study demonstrates that gallium nitrate can inhibit the activity of MMPs and may be useful as a modulator of inflammation in arthritis.

  2. Structural differences of matrix metalloproteinases. Homology modeling and energy minimization of enzyme-substrate complexes

    DEFF Research Database (Denmark)

    Terp, G E; Christensen, I T; Jørgensen, Flemming Steen

    2000-01-01

    Matrix metalloproteinases are extracellular enzymes taking part in the remodeling of extracellular matrix. The structures of the catalytic domain of MMP1, MMP3, MMP7 and MMP8 are known, but structures of enzymes belonging to this family still remain to be determined. A general approach...... to the homology modeling of matrix metalloproteinases, exemplified by the modeling of MMP2, MMP9, MMP12 and MMP14 is described. The models were refined using an energy minimization procedure developed for matrix metalloproteinases. This procedure includes incorporation of parameters for zinc and calcium ions...... in the AMBER 4.1 force field, applying a non-bonded approach and a full ion charge representation. Energy minimization of the apoenzymes yielded structures with distorted active sites, while reliable three-dimensional structures of the enzymes containing a substrate in active site were obtained. The structural...

  3. Matrix metalloproteinase inhibition reduces adventitial thickening and collagen accumulation following balloon dilation

    NARCIS (Netherlands)

    Sierevogel, MJ; Velema, E; van der Meer, FJ; Nijhuis, MO; de Kleijn, DPV; Borst, C; Pasterkamp, G

    2002-01-01

    Objective: Constrictive arterial remodeling following balloon angioplasty has been related to adventitial collagen accumulation and subsequent thickening and can be prevented by matrix ructalloprotemase (MMP) inhibition. Following balloon dilation, we examined the effect of MMP inhibition on colla-e

  4. Selective Allosteric Inhibition of MMP9 Is Efficacious in Preclinical Models of Ulcerative Colitis and Colorectal Cancer.

    Directory of Open Access Journals (Sweden)

    Derek C Marshall

    Full Text Available Expression of matrix metalloproteinase 9 (MMP9 is elevated in a variety of inflammatory and oncology indications, including ulcerative colitis and colorectal cancer. MMP9 is a downstream effector and an upstream mediator of pathways involved in growth and inflammation, and has long been viewed as a promising therapeutic target. However, previous efforts to target matrix metalloproteinases (MMPs, including MMP9, have utilized broad-spectrum or semi-selective inhibitors. While some of these drugs showed signs of efficacy in patients, all MMP-targeted inhibitors have been hampered by dose-limiting toxicity or insufficient clinical benefit, likely due to their lack of specificity. Here, we show that selective inhibition of MMP9 did not induce musculoskeletal syndrome (a characteristic toxicity of pan-MMP inhibitors in a rat model, but did reduce disease severity in a dextran sodium sulfate-induced mouse model of ulcerative colitis. We also found that MMP9 inhibition decreased tumor growth and metastases incidence in a surgical orthotopic xenograft model of colorectal carcinoma, and that inhibition of either tumor- or stroma-derived MMP9 was sufficient to reduce primary tumor growth. Collectively, these data suggest that selective MMP9 inhibition is a promising therapeutic strategy for treatment of inflammatory and oncology indications in which MMP9 is upregulated and is associated with disease pathology, such as ulcerative colitis and colorectal cancer. In addition, we report the development of a potent and highly selective allosteric MMP9 inhibitor, the humanized monoclonal antibody GS-5745, which can be used to evaluate the therapeutic potential of MMP9 inhibition in patients.

  5. MMP-2和MMP-9在肺癌组织中的表达%Expression of MMP-2 and MMP-9 in Lung Cancer

    Institute of Scientific and Technical Information of China (English)

    孙书明; 周妍; 罗金芳; 桂律; 胡志雄; 金盈; 金复生

    2003-01-01

    目的:研究肺癌和正常肺组织中基质金属蛋白酶2 (MMP-2)和MMP-9的表达.方法:采用免疫组织化学EnVision方法, 对71例肺癌标本和31例正常肺组织检测MMP-2和MMP-9的表达.结果:71例肺癌标本中,小细胞肺癌(SCLC)4例,MMP-2表达阳性3例(75.0%),MMP-9表达阳性2例(50.0%);非小细胞肺癌(NSCLC)有67例,年龄40岁以上64例,MMP-2表达阳性39例,阳性率60.9%(39/64);MMP-9表达阳性46例,阳性率71.9%(46/64).正常肺标本31例,年龄40岁以下MMP-2和MMP-9表达均为阴性,年龄在40岁以上26例,MMP-2阳性表达9例,阳性率34.6%(9/26),MMP-9阳性表达10例,阳性率38.5%(10/26);40岁以上NSCLC标本的MMP-2和MMP-9阳性表达与正常肺标本比较,差异均有显著意义.结论:在NSCLC中高表达的MMP-2和MMP-9均是好的肿瘤标记物,但是,MMP-9阳性率比MMP-2更高,因此MMP-9的检测具有更大的临床意义.

  6. Phosphorylation status of 72 kDa MMP-2 determines its structure and activity in response to peroxynitrite.

    Directory of Open Access Journals (Sweden)

    Anna Laura Jacob-Ferreira

    Full Text Available Matrix metalloproteinase-2 (MMP-2 is a key intra- and extra-cellular protease which contributes to several oxidative stress related pathologies. A molecular understanding of 72 kDa MMP-2 activity, directly mediated by S-glutathiolation of its cysteine residues in the presence of peroxynitrite (ONOO(- and by phosphorylation of its serine and threonine residues, is essential to develop new generation inhibitors of intracellular MMP-2. Within its propeptide and collagen binding domains there is an interesting juxtaposition of predicted phosphorylation sites with nearby cysteine residues which form disulfide bonds. However, the combined effect of these two post-translational modifications on MMP-2 activity has not been studied. The activity of human recombinant 72 kDa MMP-2 (hrMMP-2 following in vitro treatments was measured by troponin I proteolysis assay and a kinetic activity assay using a fluorogenic peptide substrate. ONOO(- treatment in the presence of 30 µM glutathione resulted in concentration-dependent changes in MMP-2 activity, with 0.1-1 µM increasing up to twofold and 100 µM attenuating its activity. Dephosphorylation of MMP-2 with alkaline phosphatase markedly increased its activity by sevenfold, either with or without ONOO(-. Dephosphorylation of MMP-2 also affected the conformational structure of the enzyme as revealed by circular dichroism studies, suggesting an increase in the proportion of α-helices and a decrease in β-strands compared to the phosphorylated form of MMP-2. These results suggest that ONOO(- activation (at low µM and inactivation (at high µM of 72 kDa MMP-2, in the presence or absence of glutathione, is also influenced by its phosphorylation status. These insights into the role of post-translational modifications in the structure and activity of 72 kDa MMP-2 will aid in the development of inhibitors specifically targeting intracellular MMP-2.

  7. A nonintrinsic regional basis for increased infrarenal aortic MMP-9 expression and activity.

    Science.gov (United States)

    Ailawadi, Gorav; Knipp, Brian S; Lu, Guanyi; Roelofs, Karen J; Ford, John W; Hannawa, Kevin K; Bishop, Keith; Thanaporn, Porama; Henke, Peter K; Stanley, James C; Upchurch, Gilbert R

    2003-05-01

    This investigation was undertaken to determine whether intrinsic or regional factors at different anatomic sites of the aorta affect expression and activity of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). Aortas from Sprague-Dawley rats (n = 22) were divided into arch, descending thoracic, and infrarenal abdominal segments. Specimens were stimulated with interleukin-1beta (IL-1beta) (2 ng/mL) for 72 hours. In separate experiments, syngeneic aortic segments were transplanted from the thoracic or abdominal aortas of donor rats into the infrarenal aortic position of recipient rats (n = 12 each). At 4 weeks, aortas from rats who had received transplants were harvested, sectioned into arch, thoracic, and transplanted thoracic or transplanted abdominal segments, and stimulated with IL-1beta. Reverse transcriptase polymerase chain reaction, zymography, and reverse zymography were performed to assess MMP-9, MMP-2, and TIMP-1 in all aortic segments. Differences were assessed with analysis of variance (ANOVA) and post-hoc Tukey test. In control rats, abdominal segments had significantly higher MMP-9 expression compared with arch and thoracic segments (P <.002). Total MMP-9 activity was also higher in abdominal segments (P <.02). In rats who received transplants, transplanted thoracic (P <.004) and transplanted abdominal (P <.05) segments demonstrated upregulation of MMP-9 expression, compared with control arch and thoracic segments. Zymography documented increased total MMP-9 activity in transplanted thoracic (P <.03) and transplanted abdominal (P <.04) segments versus arch and thoracic segments. No significant difference in MMP-9 expression was found between control abdominal, transplanted thoracic, or transplanted abdominal segments. No significant differences in MMP-2 or TIMP-1 expression or activity were demonstrated in either control or transplanted segments. These data demonstrate that variations in aortic MMP-9 expression and

  8. Homocysteine enhances MMP-9 production in murine macrophages via ERK and Akt signaling pathways

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Seung Jin; Lee, Yi Sle; Seo, Kyo Won; Bae, Jin Ung; Kim, Gyu Hee; Park, So Youn; Kim, Chi Dae, E-mail: chidkim@pusan.ac.kr

    2012-04-01

    Homocysteine (Hcy) at elevated levels is an independent risk factor of cardiovascular diseases, including atherosclerosis. In the present study, we investigated the effect of Hcy on the production of matrix metalloproteinases (MMP) in murine macrophages. Among the MMP known to regulate the activities of collagenase and gelatinase, Hcy exclusively increased the gelatinolytic activity of MMP-9 in J774A.1 cells as well as in mouse peritoneal macrophages. Furthermore, this activity was found to be correlated with Western blot findings in J774A.1 cells, which showed that MMP-9 expression was concentration- and time-dependently increased by Hcy. Inhibition of the ERK and Akt pathways led to a significant decrease in Hcy-induced MMP-9 expression, and combined treatment with inhibitors of the ERK and Akt pathways showed an additive effects. Activity assays for ERK and Akt showed that Hcy increased the phosphorylation of both, but these phosphorylation were not affected by inhibitors of the Akt and ERK pathways. In line with these findings, the molecular inhibition of ERK and Akt using siRNA did not affect the Hcy-induced phosphorylation of Akt and ERK, respectively. Taken together, these findings suggest that Hcy enhances MMP-9 production in murine macrophages by separately activating the ERK and Akt signaling pathways. -- Highlights: ► Homocysteine (Hcy) induced MMP-9 production in murine macrophages. ► Hcy induced MMP-9 production through ERK and Akt signaling pathways. ► ERK and Akt signaling pathways were activated by Hcy in murine macrophages. ► ERK and Akt pathways were additively act on Hcy-induced MMP-9 production. ► Hcy enhances MMP-9 production in macrophages via activation of ERK and Akt signaling pathways in an independent manner.

  9. In-vitro and in-vivo imaging of MMP activity in cartilage and joint injury

    Energy Technology Data Exchange (ETDEWEB)

    Fukui, Tomoaki; Tenborg, Elizabeth; Yik, Jasper H.N.; Haudenschild, Dominik R., E-mail: DRHaudenschild@ucdavis.edu

    2015-05-08

    Non-destructive detection of cartilage-degrading activities represents an advance in osteoarthritis (OA) research, with implications in studies of OA pathogenesis, progression, and intervention strategies. Matrix metalloproteinases (MMPs) are principal cartilage degrading enzymes that contribute to OA pathogenesis. MMPSense750 is an in-vivo fluorimetric imaging probe with the potential to continuously and non-invasively trace real-time MMP activities, but its use in OA-related research has not been reported. Our objective is to detect and characterize the early degradation activities shortly after cartilage or joint injury with MMPSense750. We determined the appropriate concentration, assay time, and linear range using various concentrations of recombinant MMPs as standards. We then quantified MMP activity from cartilage explants subjected to either mechanical injury or inflammatory cytokine treatment in-vitro. Finally, we performed in-vivo MMP imaging of a mouse model of post-traumatic OA. Our in-vitro results showed that the optimal assay time was highly dependent on the MMP enzyme. In cartilage explant culture media, mechanical impact or cytokine treatment increased MMP activity. Injured knees of mice showed significantly higher fluorescent signal than uninjured knees. We conclude that MMPSense750 detects human MMP activities and can be used for in-vitro study with cartilage, as well as in-vivo studies of knee injury, and can offering real-time insight into the degradative processes that occurring within the joint before structural changes become evident radiographically. - Highlights: • MMPSense750 is near-infrared fluorescent probe which can detect MMP activity. • MMPSense750 can detect human MMP-3, -9, and -13. • The reaction kinetics with MMPSense750 were different for the three MMPs. • MMPSense750 can visualized real time MMP activity in mouse injured knees. • MMPSense750 is convenient tool to evaluate real-time MMP activity non-invasively.

  10. Structural analysis and promoter characterization of the human collagenase-3 gene (MMP13)

    Energy Technology Data Exchange (ETDEWEB)

    Pendas, A.M.; Balbin, M.; Llano, E. [Universidad de Oviedo (Spain)] [and others

    1997-03-01

    Human collagenase-3 (MMP13) is a recently identified member of the matrix metalloproteinase (MMP) family that is expressed in breast carcinomas and in articular cartilage from arthritic patients. In this work we have isolated and characterized genomic clones coding for human collagenase-3. This gene is composed of 10 exons and 9 introns and spans over 12.5 kb. The overall organization of the collagenase-3 gene is similar to that of other MMP genes clustered at chromosome 11q22, including fibroblast collagenase (MMP-1), matrilysin (MMP-7), and macrophage metalloelastase (MMP-12), but is more distantly related to genes coding for stromelysin-3 (MMP-11), gelatinase-A (MMP-2), and gelatinase-B (MMP-9), which map outside of this gene cluster. Nucleotide sequence analysis of about 1 kb of the 5{prime}-flanking region of the collagenase-3 gene revealed the presence of a TATA box, an AP-1 motif, a PEA-3 consensus sequence, an osteoblast specific element (OSE-2), and a TGF-{beta} inhibitory element. Transient transfection experiments in HeLa and COS-1 cells with chloramphenicol acetyltransferase (CAT)-containing constructs showed that the AP-1 site is functional and responsible for the observed inducibility of the reporter gene by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). However, and in contrast to other MMP genes, no significative synergistic effect on CAT activity between the AP-1 and PEA-3 elements found in the collagenase-3 gene promoter was found. DNA binding analysis with nuclear extracts from HeLa cells revealed the formation of specific complexes between collagenase-3 promoter sequences containing the AP-1 site and nuclear proteins. The presence of this AP-1 functional site, which is able to confer responsiveness to a variety of tumor promoters and oncogene products, may contribute to explaining the high-level expression of collagenase-3 in breast carcinomas and degenerative joint diseases. 48 refs., 5 figs., 2 tabs.

  11. The anti-MMP activity of benzalkonium chloride

    Science.gov (United States)

    Tezvergil-Mutluay, Arzu; Mutluay, M. Murat; Gu, Li-sha; Zhang, Kai; Agee, Kelli A.; Carvalho, Ricardo M.; Manso, Adriana; Carrilho, Marcela; Tay, Franklin R.; Breschi, Lorenzo; Suh, Byoung-In; Pashley, David H.

    2013-01-01

    SUMMARY Objective This study evaluated the ability of benzalkonium chloride (BAC) to bind to dentine and to inhibit soluble recombinant MMPs and bound dentine matrix metalloproteinases (MMPs). Methods Dentine powder was prepared from extracted human molars. Half was left mineralized; the other half was completely demineralized. The binding of BAC to dentine powder was followed by measuring changes in the supernatant concentration using UV spectrometry. The inhibitory effects of BAC on rhMMP-2, -8 and -9 were followed using a commercially available in vitro proteolytic assay. Matrix-bound endogenous MMP-activity was evaluated in completely demineralized beams. Each beam was either dipped into BAC and then dropped into 1 mL of a complete medium (CM) or they were placed in 1 mL of CM containing BAC for 30 d. After 30 d, changes in the dry mass of the beams or in the hydroxyproline (HYP) content of hydrolyzates of the media were quantitated as indirect measures of matrix collagen hydrolysis by MMPs. Results Demineralized dentine powder took up 10-times more BAC than did mineralized powder. Water rinsing removed about 50% of the bound BAC, while rinsing with 0.5 M NaCl removed more than 90% of the bound BAC. BAC concentrations 0.5 wt% produced 100% inhibition of soluble recombinant MMP-2, -8 or -9, and inhibited matrix-bound MMPs between 55-66% when measured as mass loss or 76-81% when measured as solubilization of collagen peptide fragments. Conclusions BAC is effective at inhibiting both soluble recombinant MMPs and matrix-bound dentine MMPs in the absence of resins. PMID:20951183

  12. The role of hypoxia inducible factor-1α in the increased MMP-2 and MMP-9 production by human monocytes exposed to nickel nanoparticles

    Science.gov (United States)

    WAN, RONG; MO, YIQUN; CHIEN, SUFAN; LI, YIHUA; LI, YIXIN; TOLLERUD, DAVID J.; ZHANG, QUNWEI

    2016-01-01

    Nickel is an important economic commodity, but it can cause skin sensitization and may cause lung diseases such as lung fibrosis, pneumonitis, bronchial asthma and lung cancer. With development of nanotechnology, nano-sized nickel (Nano-Ni) and nano-sized titanium dioxide (Nano-TiO2) particles have been developed and produced for many years with new formulations and surface properties to meet novel demands. Our previous studies have shown that Nano-Ni instilled into rat lungs caused a greater inflammatory response as compared with standard-sized nickel (5 μm) at equivalent mass concentrations. Nano-Ni caused a persistent high level of inflammation in lungs even at low doses. Recently, several studies have shown that nanoparticles can translocate from the lungs to the circulatory system. To evaluate the potential systemic effects of metal nanoparticles, we compared the effects of Nano-Ni and Nano-TiO2 on matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) gene expression and activity. Our results showed that exposure of human monocyte U937 to Nano-Ni caused dose- and time- dependent increase in MMP-2 and MMP-9 mRNA expression and pro-MMP-2 and pro-MMP-9 activity, but Nano-TiO2 did not. Nano-Ni also caused dose- and time- related increase in tissue inhibitor of metalloproteinases 1 (TIMP-1), but Nano-TiO2 did not. To determine the potential mechanisms involved, we measured the expression of hypoxia inducible factor 1α (HIF-1α) in U937 cells exposed to Nano-Ni and Nano-TiO2. Our results showed that exposure to Nano-Ni caused HIF-1α accumulation in the nucleus. Furthermore, pre-treatment of U937 cells with heat shock protein 90 (Hsp90) inhibitor, 17-(Allylamino)-17-demethoxygeldanamycin (17-AAG), prior to exposure to Nano-Ni significantly abolished Nano-Ni-induced MMP-2 and MMP-9 mRNA upregulation and increased pro-MMP-2 and pro-MMP-9 activity. Our results suggest that HIF-1α accumulation may be involved in the increased MMP-2 and MMP-9 production in U937 cells

  13. The role of MT2-MMP in cancer progression

    Energy Technology Data Exchange (ETDEWEB)

    Ito, Emiko [Department of Molecular Pathology, Graduate School of Medicine and Health Sciences, Osaka University, Suita, Osaka 565-0871 (Japan); Yana, Ikuo [Department of Molecular Pathology, Graduate School of Medicine and Health Sciences, Osaka University, Suita, Osaka 565-0871 (Japan); Takeda Pharmaceutical Co. Ltd., Japan Development Center, Osaka 540-8645 (Japan); Fujita, Chisato; Irifune, Aiko; Takeda, Maki; Madachi, Ayako; Mori, Seiji; Hamada, Yoshinosuke; Kawaguchi, Naomasa [Department of Molecular Pathology, Graduate School of Medicine and Health Sciences, Osaka University, Suita, Osaka 565-0871 (Japan); Matsuura, Nariaki, E-mail: Matsuura@sahs.med.osaka-u.ac.jp [Department of Molecular Pathology, Graduate School of Medicine and Health Sciences, Osaka University, Suita, Osaka 565-0871 (Japan)

    2010-03-05

    The role of MT2-MMP in cancer progression remains to be elucidated in spite of many reports on MT1-MMP. Using a human fibrosarcoma cell, HT1080 and a human gastric cancer cell, TMK-1, endogenous expression of MT1-MMP or MT2-MMP was suppressed by siRNA induction to examine the influence of cancer progression in vitro and in vivo. In HT1080 cells, positive both in MT1-MMP and MT2-MMP, the migration as well as the invasion was impaired by MT1-MMP or MT2-MMP suppression. Also cell proliferation in three dimensional (3D) condition was inhibited by MT1-MMP or MT2-MMP suppression and tumor growth in the nude mice transplanted with tumor cells were reduced either MT1-MMP or MT2-MMP suppression with a prolongation of survival time in vivo. MT2-MMP suppression induces more inhibitory effects on 3D proliferation and in vivo tumor growth than MT1-MMP. On the other hand, TMK-1 cells, negative in MT1-MMP and MMP-2 but positive in MT2-MMP, all the migratory, invasive, and 3D proliferative activities in TMK-1 are decreased only by MT2-MMP suppression. These results indicate MT2-MMP might be involved in the cancer progression more than or equal to MT1-MMP independently of MMP-2 and MT1-MMP.

  14. EXPRESSION AND SIGNIFICANCE OF THE NOVEL HYPERPLASIC SUPPRESS GENE IN THE PROCESS OF BREAST CANCER%HSG、MMP-2和MMP-9在乳腺癌组织中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    夏庆安; 付玉环; 姜广建

    2012-01-01

    [目的]探讨新的增殖抑制基因(hyperplasic suppressor gene,HSG)和基质金属蛋白酶-2、9(matrix metalloproteinase-2,-9 MMP-2,9)在乳腺癌组织中的表达及意义.[方法]应用免疫组化方法检测唐山市工人医院52例乳腺癌标本和18例乳腺纤维腺瘤标本中HSG、MMP-2和MMP-9表达.[结果]52例乳腺癌组织中HSG、MMP-2和MMP-9的表达分别为46.2% (24/52)、73.1% (38/52)和63.5% (33/52),在乳腺纤维腺瘤中,MMP-2和MMP-9着色很少,HSG的着色很深,表明三者与肿瘤的发生有关;MMP-2的表达与肿瘤的TNM分期、淋巴结转移和预后复发有关(P<005);乳腺癌组HSG低表达和MMP-9高表达,与淋巴结转移有关(P<005),且HSG和MMP-2、MMP-9呈负相关(r=-0.27,P< 0.05).[结论]乳腺癌组织中MMP-2高表达预示着病人预后更差,比MMP-9对乳腺癌与后的判断更有价值.综合检测HSG、MMP-2和MMP-9的表达更有利于了解乳腺癌转移的作用机制,更好地判断乳腺癌的进展及预后.%[Objective] To determine the expression and significance of the novel hyperplasic suppress gene (HSG) and matrix metaUoproteinase-2, -9 (MMP-2, -9) in the process of breast cancer. [Methods] Iramunohistochemistry method was used to measure the expression of HSG and raetalIoproteinase-2, -9, in which 52 cases were breast cancer samples, 18 cases were breast Cbroadenoma samples. All these cases were collected by the Department of pathology of the Tangshan Worker/s Hospital. [Resufts] The positive rate of HSG, MMP-2, -9 was 46.2% (24/52), 73.1% (38/52) and 65.3% (33/52) in breast cancer tissues, respectively- A very weak Btaining of MMP-2, -9 was observed in only a few epidielia in surrounding background breast tissue and flbrooadenoma tissue, but observed a very strong staining of HSG. The presence of HSG, MMP-2, -9 was significantly greater in breast cancer tissues compared with all other breast tissues; The high leve of MMP-2 was associated with the tumor grade, lymph

  15. Matrix metalloproteinase expression and localization in turkey (Meleagris gallopavo) during the endochondral ossification process.

    Science.gov (United States)

    Simsa, S; Genina, O; Ornan, E Monsonego

    2007-06-01

    Vertebrate long bones are formed by endochondral ossification, a process accompanied by changes in extracellular matrix synthesis and remodeling, performed mainly by the matrix metalloproteinases (MMP). The temporal/spatial expression patterns of 5 members of the MMP family known to be important for endochondral ossification were studied, for the first time, in the turkey growth plate during embryonic and juvenile stages. The expression of MMP-2 was detected in the proliferative zone, MMP-3, MMP-9, and MMP-13 in cells lining the blood vessels; MMP-13 was also detected in hypertrophic chondrocytes. The MMP-16 expression was detected in the reserve zone of the growth plate. These results present a detailed survey of turkey MMP, serving as a data source (atlas) for further studies in this subject.

  16. Multiple myeloma–derived MMP-13 mediates osteoclast fusogenesis and osteolytic disease

    Science.gov (United States)

    Li, Shirong; Feng, Rentian; Ma, Huihui; Sabeh, Farideh; Roodman, G. David; Wang, Ji; Robinson, Samuel; Guo, X. Edward; Lund, Thomas; Normolle, Daniel; Mapara, Markus Y.; Weiss, Stephen J.

    2016-01-01

    Multiple myeloma (MM) cells secrete osteoclastogenic factors that promote osteolytic lesions; however, the identity of these factors is largely unknown. Here, we performed a screen of human myeloma cells to identify pro-osteoclastogenic agents that could potentially serve as therapeutic targets for ameliorating MM-associated bone disease. We found that myeloma cells express high levels of the matrix metalloproteinase MMP-13 and determined that MMP-13 directly enhances osteoclast multinucleation and bone-resorptive activity by triggering upregulation of the cell fusogen DC-STAMP. Moreover, this effect was independent of the proteolytic activity of the enzyme. Further, in mouse xenograft models, silencing MMP-13 expression in myeloma cells inhibited the development of osteolytic lesions. In patient cohorts, MMP-13 expression was localized to BM-associated myeloma cells, while elevated MMP-13 serum levels were able to correctly predict the presence of active bone disease. Together, these data demonstrate that MMP-13 is critical for the development of osteolytic lesions in MM and that targeting the MMP-13 protein — rather than its catalytic activity — constitutes a potential approach to mitigating bone disease in affected patients. PMID:27043283

  17. Identification of MMP-2 as a novel enhancer of cerebellar granule cell proliferation.

    Science.gov (United States)

    Verslegers, Mieke; Van Hove, Inge; Buyens, Tom; Dekeyster, Eline; Knevels, Ellen; Moons, Lieve

    2013-11-01

    During the first postnatal days in the mouse, granule cells (GCs) undergo massive proliferation, which then gradually decreases. Matrix metalloproteinase-2 (MMP-2), a Zn(2+)-dependent proteolytic enzyme, is involved in a wide variety of pathological and physiological pathways. Evidence for a role of this proteinase in cell proliferation is emerging, reporting its involvement in pathological proliferation, as well as during neurogenesis and developmental proliferation of non-CNS tissues. In this study, MMP-2 protein expression was observed within the early postnatal cerebellar cortex, predominantly in Purkinje cells and within the GC proliferative zone, i.e. the superficial external granular layer (EGL). Consistently, the spatiotemporal MMP-2 mRNA and protein profiles highly correlated with the peak of GC precursor (GCP) proliferation and detailed morphometric analyses of MMP-2 deficient cerebella revealed a thinner EGL due to a decreased GCP proliferation. BrdU cumulative experiments, performed to measure the length of different cell cycle phases, further disclosed a transiently prolonged S-phase in MMP-2 deficient GCPs during early cerebellar development. In consequence, MMP-2 deficient animals displayed a transient delay in GC migration towards the IGL. In conclusion, our findings provide important evidence for a role for MMP-2 in neuronal proliferation and cell cycle kinetics in the developing CNS.

  18. Mmp1 processing of the PDF neuropeptide regulates circadian structural plasticity of pacemaker neurons.

    Science.gov (United States)

    Depetris-Chauvin, Ana; Fernández-Gamba, Agata; Gorostiza, E Axel; Herrero, Anastasia; Castaño, Eduardo M; Ceriani, M Fernanda

    2014-10-01

    In the Drosophila brain, the neuropeptide PIGMENT DISPERSING FACTOR (PDF) is expressed in the small and large Lateral ventral neurons (LNvs) and regulates circadian locomotor behavior. Interestingly, PDF immunoreactivity at the dorsal terminals changes across the day as synaptic contacts do as a result of a remarkable remodeling of sLNv projections. Despite the relevance of this phenomenon to circuit plasticity and behavior, the underlying mechanisms remain poorly understood. In this work we provide evidence that PDF along with matrix metalloproteinases (Mmp1 and 2) are key in the control of circadian structural remodeling. Adult-specific downregulation of PDF levels per se hampers circadian axonal remodeling, as it does altering Mmp1 or Mmp2 levels within PDF neurons post-developmentally. However, only Mmp1 affects PDF immunoreactivity at the dorsal terminals and exerts a clear effect on overt behavior. In vitro analysis demonstrated that PDF is hydrolyzed by Mmp1, thereby suggesting that Mmp1 could directly terminate its biological activity. These data demonstrate that Mmp1 modulates PDF processing, which leads to daily structural remodeling and circadian behavior.

  19. PGE2 reduces MMP-14 and increases plasminogen activator inhibitor-1 in cardiac fibroblasts.

    Science.gov (United States)

    Kassem, Kamal M; Clevenger, Margarette H; Szandzik, David L; Peterson, Edward; Harding, Pamela

    2014-10-01

    Prostaglandin E2 (PGE2) is elevated during cardiac injury and we have previously shown that mice lacking the PGE2 EP4 receptor display dilated cardiomyopathy (DCM) with increased expression of the membrane type matrix metalloproteinase, MMP-14. We thus hypothesized that PGE2 regulates expression of MMP-14 and also affects fibroblast migration. Primary cultures of neonatal rat ventricular fibroblasts (NVFs) were used to test the effects of PGE2. Gene and protein expression was assessed by real time RT-PCR and Western blot, MMP activity was determined by zymography and migration of NVF was assessed by motility in a transwell system. PGE2 reduced expression of MMP-14 and these effects were antagonized by an EP4 antagonist. An EP4 agonist mimicked the effect of PGE2. PGE2 also increased mRNA and protein levels of plasminogen activator inhibitor-1 (PAI-1), an inhibitor of MMP activation. However, PGE2-stimulation of PAI-1 was mediated by the EP1/EP3 receptor and not EP4. Migration of NVF was assessed by motility in a transwell system. Treatment of NVFs with PGE2 reduced the number of cells migrating toward 10% FCS. Treatment with the EP2 agonist also reduced migration but did not affect MMP-14 expression or PAI-1. Our results suggest that PGE2 utilizes different receptors and mechanisms to ultimately decrease MMP expression and NVF migration. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Mmp1 processing of the PDF neuropeptide regulates circadian structural plasticity of pacemaker neurons.

    Directory of Open Access Journals (Sweden)

    Ana Depetris-Chauvin

    2014-10-01

    Full Text Available In the Drosophila brain, the neuropeptide PIGMENT DISPERSING FACTOR (PDF is expressed in the small and large Lateral ventral neurons (LNvs and regulates circadian locomotor behavior. Interestingly, PDF immunoreactivity at the dorsal terminals changes across the day as synaptic contacts do as a result of a remarkable remodeling of sLNv projections. Despite the relevance of this phenomenon to circuit plasticity and behavior, the underlying mechanisms remain poorly understood. In this work we provide evidence that PDF along with matrix metalloproteinases (Mmp1 and 2 are key in the control of circadian structural remodeling. Adult-specific downregulation of PDF levels per se hampers circadian axonal remodeling, as it does altering Mmp1 or Mmp2 levels within PDF neurons post-developmentally. However, only Mmp1 affects PDF immunoreactivity at the dorsal terminals and exerts a clear effect on overt behavior. In vitro analysis demonstrated that PDF is hydrolyzed by Mmp1, thereby suggesting that Mmp1 could directly terminate its biological activity. These data demonstrate that Mmp1 modulates PDF processing, which leads to daily structural remodeling and circadian behavior.

  1. Homozygous and compound heterozygous MMP20 mutations in amelogenesis imperfecta.

    Science.gov (United States)

    Gasse, B; Karayigit, E; Mathieu, E; Jung, S; Garret, A; Huckert, M; Morkmued, S; Schneider, C; Vidal, L; Hemmerlé, J; Sire, J-Y; Bloch-Zupan, A

    2013-07-01

    In this article, we focus on hypomaturation autosomal-recessive-type amelogenesis imperfecta (type IIA2) and describe 2 new causal Matrix metalloproteinase 20 (MMP20) mutations validated in two unrelated families: a missense mutation p.T130I at the expected homozygous state, and a compound heterozygous mutation having the same mutation combined with a nucleotide deletion, leading to a premature stop codon (p.N120fz*2). We characterized the enamel structure of the latter case using scanning electron microscopy analysis and microanalysis (Energy-dispersive X-ray Spectroscopy, EDX) and confirmed the hypomaturation-type amelogenesis imperfecta as identified in the clinical diagnosis. The mineralized content was slightly decreased, with magnesium substituting for calcium in the crystal structure. The anomalies affected enamel with minimal inter-rod enamel present and apatite crystals perpendicular to the enamel prisms, suggesting a possible new role for MMP20 in enamel formation.

  2. Peak in matrix metaloproteinases-2 levels observed during recovery from olfactory nerve injury.

    Science.gov (United States)

    Costanzo, Richard M; Perrino, Lisa A

    2008-02-12

    Matrix metalloproteineases are associated with extracellular remodeling that occurs in injury and repair processes in the central nervous system (CNS). We examined the role of MMP-2 in a model of olfactory nerve injury and found that MMP-2 levels increased several hours following injury, peaked at day 7 and then decreased rapidly. We previously reported a rapid increase in MMP-9, within 5 h after nerve injury, corresponding to neuronal degeneration and increased glial activity. In this study, we show that MMP-2 peaks later than MMP-9, at the onset of neuronal regeneration and repair. Using MMP-9 knockout mice, we determined that the MMP-2 increase is independent of MMP-9. Our data suggest that MMP-2 and MMP-9 may play different roles in the injury and repair processes.

  3. MMP2 and MMP9 serum levels are associated with favorable outcome in patients with inflammatory breast cancer treated with bevacizumab-based neoadjuvant chemotherapy in the BEVERLY-2 study

    Science.gov (United States)

    Tabouret, Emeline; Bertucci, François; Pierga, Jean-Yves; Petit, Thierry; Levy, Christelle; Ferrero, Jean-Marc; Campone, Mario; Gligorov, Joseph; Lerebours, Florence; Roché, Henri; Bachelot, Thomas; van Laere, Steven; Ueno, Naoto T.; Toiron, Yves; Finetti, Pascal; Birnbaum, Daniel; Borg, Jean-Paul; Viens, Patrice

    2016-01-01

    Purpose Addition of bevacizumab to trastuzumab-based neoadjuvant chemotherapy in HER2-positive inflammatory breast cancer (IBC) was associated with favorable outcome in the BEVERLY-2 phase II trial. Circulating levels of matrix metalloproteinases (MMP) 2 and 9 were correlated to high response rate and prolonged survival in high-grade glioma treated with bevacizumab. We examined the prognostic impact of MMP2 and MMP9 serum levels in BEVERLY-2 patients. Experimental design MMP2 and MMP9 serum levels were assessed using ELISA at baseline and before surgery in 45/52 available samples. Correlations were tested with pathological complete response (pCR), disease-free survival (DFS) and overall survival (OS). Results Baseline (b) MMP2 and MMP9 serum levels were independent from patient characteristics and circulating tumor or endothelial cells, and were not correlated to pCR. High bMMP2 was correlated to better DFS (p=0.001) and OS (p=0.032), while low bMMP9 was correlated to better OS (p=0.022) and tended to be associated with longer DFS (p=0.071). In multivariate analyses, bMMP2 (p=0.003, Hazard Ratio [HR]: 0.115) and bMMP9 (p=0.041, HR: 3.511) remained correlated to DFS. As continuous variables, bMMP2 was associated with relapse (p=0.002) and death (p=0.049), while bMMP9 was associated with death (p=0.035). During treatment, significant increase in MMP2 and decrease in MMP9 levels (p<0.001 for both) were observed in 100% and 87% of patients respectively. Conclusions High bMMP2 and low bMMP9 serum levels were associated with better survival in HER2-positive IBC patients treated with bevacizumab- and trastuzumab-based neoadjuvant chemotherapy. Their predictive value of bevacizumab benefit should be evaluated in a randomized trial. PMID:26921265

  4. The Effects of Amphiregulin Induced MMP-13 Production in Human Osteoarthritis Synovial Fibroblast

    Directory of Open Access Journals (Sweden)

    Yi-Te Chen

    2014-01-01

    Full Text Available Osteoarthritis (OA belongs to a group of degenerative diseases. Synovial inflammation, cartilage abrasion, and subchondral sclerosis are characteristics of OA. Researchers do not fully understand the exact etiology of OA. However, matrix metalloproteinases (MMPs, which are responsible for cartilage matrix degradation, play a pivotal role in the progression of OA. Amphiregulin (AREG binds to the EGF receptor (EGFR and activates downstream proteins. AREG is involved in a variety of pathological processes, such as the development of tumors, inflammatory diseases, and rheumatoid arthritis. However, the relationship between AREG and MMP-13 in OA synovial fibroblasts (SFs remains unclear. We investigated the signaling pathway involved in AREG-induced MMP-13 production in SFs. AREG caused MMP-13 production in a concentration- and time-dependent manner. The results of using pharmacological inhibitors and EGFR siRNA to block EGFR revealed that the EGFR receptor was involved in the AREG-mediated upregulation of MMP-13. AREG-mediated MMP-13 production was attenuated by PI3K and Akt inhibitors. The stimulation of cells by using AREG activated p65 phosphorylation and p65 translocation from the cytosol to the nucleus. Our results provide evidence that AREG acts through the EGFR and activates PI3K, Akt, and finally NF-kappaB on the MMP-13 promoter, thus contributing to cartilage destruction during osteoarthritis.

  5. Matrix metalloproteinase 20-dentin sialophosphoprotein interaction in oral cancer.

    Science.gov (United States)

    Saxena, G; Koli, K; de la Garza, J; Ogbureke, K U E

    2015-04-01

    Matrix metalloproteinase 20 (MMP-20), widely regarded as tooth specific, participates with MMP-2 in processing dentin sialophosphoprotein (DSPP) into dentin sialoprotein, dentin phosphoprotein, and dentin glycoprotein. In biochemical system, MMP-2, MMP-3, and MMP-9 bind with high affinity to, and are activated by, specific small integrin-binding ligand N-linked glycoproteins (SIBLINGs): bone sialoprotein, osteopontin, and dentin matrix protein 1, respectively. Subsequent reports documented possible biological relevance of SIBLING-MMP interaction in vivo by showing that SIBLINGs are always coexpressed with their MMP partners. However, the cognate MMPs for 2 other SIBLINGs-DSPP and matrix extracellular phosphogylcoprotein-are yet to be identified. Our goal was to investigate MMP-20 expression and to explore preliminary evidence of its interaction with DSPP in oral squamous cell carcinomas (OSCCs). Immunohistochemistry analysis of sections from 21 cases of archived human OSCC tissues showed immunoreactivity for MMP-20 in 18 (86%) and coexpression with DSPP in all 15 cases (71%) positive for DSPP. Similarly, 28 (93%) of 30 cases of oral epithelial dysplasia were positive for MMP-20. Western blot and quantitative real-time polymerase chain reaction analysis on OSCC cell lines showed upregulation of MMP-20 protein and mRNA, respectively, while immunofluorescence showed coexpression of MMP-20 and DSPP. Colocalization and potential interaction of MMP-20 with dentin sialoprotein was confirmed by coimmunoprecipitation and mass spectrometry analysis of immunoprecipitation product from OSCC cell lysate, and in situ proximity ligation assays. Significantly, results of chromatin immunoprecipation revealed a 9-fold enrichment of DSPP at MMP-20 promoter-proximal elements. Our data provide evidence that MMP-20 has a wider tissue distribution than previously acknowledged. MMP-20-DSPP specific interaction, excluding other MMP-20-SIBLING pairings, identifies MMP-20 as DSPP cognate MMP

  6. Rock2 promotes the invasion and metastasis of hepatocellular carcinoma by modifying MMP2 ubiquitination and degradation.

    Science.gov (United States)

    Huang, Da; Du, Xiaohong; Yuan, Rongfa; Chen, Leifeng; Liu, Tiande; Wen, Chongyu; Huang, Mingwen; Li, Ming; Hao, Liang; Shao, Jianghua

    2014-10-10

    Rho-associated coiled-coil-containing protein kinase 2 (Rock2) is a downstream effector of Rho that plays an important role in the tumorigenesis and progression of hepatocellular carcinoma (HCC). Matrix metalloproteinase 2 (MMP2) is a master regulator of tumor metastasis. In this study, we investigated the collections of Rock2 and MMP2 in HCCs and determined the potential role and molecular mechanism of Rock2 in MMP2-mediated invasiveness and metastasis. We found that Rock2 and MMP2 were markedly overexpressed in HCCs compared with the corresponding adjacent tissues, where a positive correlation in their expression was found. The knockdown of Rock2 significantly decreased MMP2 expression and inhibited the invasion and metastasis of HCC in vitro and in vivo. Additionally, the upregulation of MMP2 rescued the decreased migration and invasion induced by the knockdown of Rock2, whereas the knockdown of MMP2 decreased Rock2-enhanced HCC migration and invasion. Mechanistically, Rock2 stabilized MMP2 by preventing its ubiquitination and degradation. Together, our results link two drivers of invasion and metastasis in HCC and identify a novel pathway for MMP2 control.

  7. Implications of MMP9 for Blood Brain Barrier Disruption And Hemorrhagic Transformation Following Ischemic Stroke

    Directory of Open Access Journals (Sweden)

    Renee Jade Turner

    2016-03-01

    Full Text Available Numerous studies have documented increases in matrix metalloproteinases (MMPs, specifically MMP-9 levels following stroke, with such perturbations associated with disruption of the blood brain barrier (BBB, increased risk of hemorrhagic complications and worsened outcome. Despite this, controversy remains as to which cells release MMP-9 at the normal and pathological BBB, with even less clarity in the context of stroke. This may be further complicated by the influence of tissue plasminogen activator (tPA treatment. The aim of the present review is to examine the relationship between neutrophils, MMP-9 and tPA following ischemic stroke to elucidate which cells are responsible for the increases in MMP-9 and resultant barrier changes and hemorrhage observed following stroke.

  8. Implications of MMP9 for Blood Brain Barrier Disruption and Hemorrhagic Transformation Following Ischemic Stroke

    Science.gov (United States)

    Turner, Renée J.; Sharp, Frank R.

    2016-01-01

    Numerous studies have documented increases in matrix metalloproteinases (MMPs), specifically MMP-9 levels following stroke, with such perturbations associated with disruption of the blood brain barrier (BBB), increased risk of hemorrhagic complications, and worsened outcome. Despite this, controversy remains as to which cells release MMP-9 at the normal and pathological BBB, with even less clarity in the context of stroke. This may be further complicated by the influence of tissue plasminogen activator (tPA) treatment. The aim of the present review is to examine the relationship between neutrophils, MMP-9 and tPA following ischemic stroke to elucidate which cells are responsible for the increases in MMP-9 and resultant barrier changes and hemorrhage observed following stroke. PMID:26973468

  9. Synthesis of bicyclic molecular scaffolds (BTAa): an investigation towards new selective MMP-12 inhibitors.

    Science.gov (United States)

    Mannino, Claudia; Nievo, Marco; Machetti, Fabrizio; Papakyriakou, Athanasios; Calderone, Vito; Fragai, Marco; Guarna, Antonio

    2006-11-15

    Starting from 3-aza-6,8-dioxa-bicyclo[3.2.1]octane scaffold (BTAa) a virtual library of molecules was generated and screened in silico against the crystal structure of the Human Macrophage Metalloelastase (MMP-12). The molecules obtaining high score were synthesized and the affinity for the catalytic domain of MMP-12 was experimentally proved by NMR experiments. A BTAa scaffold 20 having a N-hydroxyurea group in position 3 and a p-phenylbenzylcarboxy amide in position 7 showed a fair inhibition potency (IC50 = 149 microM) for MMP-12 and some selectivity towards five different MMPs. These results, taken together with the X-ray structure of the adduct between MMP-12, the inhibitor 20 and the acetohydroxamic acid (AHA), suggest that bicyclic scaffold derivatives may be exploited for the design of new selective matrix metalloproteinase inhibitors (MMPIs).

  10. Differential expression of MMP-2, MMP-9 and TIMP proteins in thoracic aortic aneurysm - comparison with and without bicuspid aortic valve: a meta-analysis.

    Science.gov (United States)

    Rabkin, Simon W

    2014-11-01

    Hintergrund: Es wird angenommen, dass das Gleichgewicht der Matrix- Metalloproteasen (MMPs) und der Gewebeinhibitoren von Metalloproteinasen (TIMPs ) in der Aorta eine entscheidende Rolle bei der Ausbildung von Aneurysmen spielt. Das Ziel dieser Studie war es, eine Meta-Analyse von Studien durchzuführen, die die Protein-Expression von MMPs und TIMPs bei Aneurysmen der Aorta ascendens (TAA) untersuchten. Dabei sollte zwischen trikuspiden (TAV) und bikuspiden Aortenklappen (BAV) unterschieden werden. Methoden: Medline und Embase OvidSP wurden systematisch nach Studien abgefragt, die MMPs oder TIMPs aus TAAs und Kontrollen untersuchten. Eine entsprechende Suche erfolgte zum Vergleich zwischen BAV und TAV. Ergebnisse: Acht Studien erfüllten die Einschlusskriterien. Es gab einen signifikanten Anstieg der MMP-9 und keine Veränderung in MMP-2 in der Aorta von Personen mit TAA (N = 106 ) im Vergleich zur Kontrolle (n = 30). Zudem gab es eine hochsignifikante Verringerung der TIMP-1 und TIMP-2 in TAA (N = 93) im Vergleich zur Kontrolle (n = 24 ), was zu einem MMP-9 zu TIMP-1 oder TIMP-2-Verhältnis von mehr als 3,5 im Vergleich zu Kontrollen führte. Es fand sich eine hochsignifikante Erhöhung der MMP-2, aber nicht der MMP-9 in TAAs mit BAV (N = 112 ) im Vergleich zu TAV (N = 53). Es gab eine signifikante Reduktion von TIMP-1 bei BAV im Vergleich zu TAV, aber keine Änderung von TIMP-2, TIMP-3 oder TIMP-4 . Schlussfolgerungen: Diese Daten deuten darauf hin, dass MMP eine Rolle in der Pathogenese von TAA spielt. Es gibt eine unterschiedliche Expression mit erhöhtem MMP-9 und verminderten TIMP-1 und -2 bei den häufigsten Formen des TAA. MMP-2 ist erhöht und nur TIMP-1 vermindert bei TAA mit BAV im Vergleich zu TAV.

  11. Effect of Polypeptide Extract from Scorpion Venom on MMP2 and MMP9 Expression in Leukemia -NOD/SCID Mice%蝎毒多肽提取物对白血病NOD/SCID小鼠MMP2、MMP9表达的影响

    Institute of Scientific and Technical Information of China (English)

    杨文华; 郝征; 杨向东; 史哲新; 于文俊; 吕俊秀

    2009-01-01

    Objective: To investigate the effect of polypeptide extract from scorpion venom (PESV) on the matrix metalloproteinase2(MMP2)and matrix metalloproteinase9(MMP9)expression in leukemia-NOD/SCID mice, and the intervention mechanism of PESV in the multiplication and infiltration of leukemic cells thereof. Methods: In order to establish the animal model of outer marrow infiltration of human leukemia,bone marrow mononuclear cells of leukemia patients, irradiated 270 cGy on body by ~(137)Cs, were injected into NOD/SCID mice. The mice were randomly divided into five groups. The groups I, II and III were treated with different concentrations of PESV. Group Ⅳ was the model group injected by the normal saline solution. GroupⅤ was taken as control. The peripheral white blood cell count and blood smear were observed in groups. All of the mice were killed after four-week observation and MMP2 and MMP9 expressions were examined using Real time PCR method. Results: The expression levels of MMP2 and MMP9 were significantly lower in group I, group II and group III than that of the model group(P < 0.05). The expression levels of MMP2 and MMP9 were related to the concentration of PESV. Moreover, the peripheral white blood cell count and blood smear were more normal in mice treated with PESV than those of mice of model group. Conclusion: PESV inhibited the overexpression of MMP2 and MMP9 in leukemia-NOD/SCID mice, which significantly inhibited the multiplication and infiltration of leukemic cells.%目的:观察蝎毒多肽提取物(PESV)对白血病NOD/SCID小鼠基质金属蛋白酶(MMP)2、MMP9表达的影响,探讨PESV对白血病细胞外基质降解和髓外浸润的干预机制.方法:首先选取急性白血病患者骨髓单个核细胞注入经过铯-137源照射的NOD/SCID小鼠体内,建立人白血病NOD/SCID小鼠髓外浸润模型;再将实验小鼠随机分组,Ⅰ组、Ⅱ组、Ⅲ组分别注射不同浓度的PESV,Ⅳ组为模型组注射生理盐

  12. Immunohistochemical analysis of MMP-9, MMP-2 and TIMP-1, TIMP-2 expression in the central nervous system following infection with viral and bacterial meningitis.

    Science.gov (United States)

    Sulik, Artur; Chyczewski, Lech

    2008-01-01

    Matrix metalloproteinases (MMPs) are capable of degrading components of the basal lamina of cerebral vessels, thereby disrupting the blood-brain barrier and inducing leukocyte recruitment. This study provides comprehensive information regarding the cell specificity of matrix metalloproteinases (MMP-2, MMP-9) and their binding tissue inhibitors (TIMP-1, TIMP-2) in the central nervous system during viral and bacterial meningitis. Specifically, we evaluated the immunoreactivity of MMPs and TIMPs in various cell types in brain parenchyma and meninges obtained from autopsy tissues. We found that a higher proportion of endothelial cells were positive for MMP-9 during meningitis when compared to controls. In addition, the immunoreactivity of MMP-9 decreased and the immunoreactivity of TIMP-1 increased in astrocytes upon infection. Furthermore, the results of this study revealed that mononuclear cells were highly immunoreactive for TIMP-1, TIMP-2 and MMP-9 during viral meningitis and that the expression of TIMPs in polymorphonuclear cells was even higher during bacterial meningitis. Taken together the results of this study indicated that the central nervous system resident cells and inflammatory infiltrates contribute to MMPs activity and that the expression patterns vary between cell types and in response to viral and bacterial meningitis.

  13. Immunohistochemical analysis of MMP-9, MMP-2 and TIMP-1, TIMP-2 expression in the central nervous system following infection with viral and bacterial meningitis.

    Directory of Open Access Journals (Sweden)

    Lech Chyczewski

    2009-01-01

    Full Text Available Matrix metalloproteinases (MMPs are capable of degrading components of the basal lamina of cerebral vessels, thereby disrupting the blood-brain barrier and inducing leukocyte recruitment. This study provides comprehensive information regarding the cell specificity of matrix metalloproteinases (MMP-2, MMP-9 and their binding tissue inhibitors (TIMP-1, TIMP-2 in the central nervous system during viral and bacterial meningitis. Specifically, we evaluated the immunoreactivity of MMPs and TIMPs in various cell types in brain parenchyma and meninges obtained from autopsy tissues. We found that a higher proportion of endothelial cells were positive for MMP-9 during meningitis when compared to controls. In addition, the immunoreactivity of MMP-9 decreased and the immunoreactivity of TIMP-1 increased in astrocytes upon infection. Furthermore, the results of this study revealed that mononuclear cells were highly immunoreactive for TIMP-1, TIMP-2 and MMP-9 during viral meningitis and that the expression of TIMPs in polymorphonuclear cells was even higher during bacterial meningitis. Taken together the results of this study indicated that the central nervous system resident cells and inflammatory infiltrates contribute to MMPs activity and that the expression patterns vary between cell types and in response to viral and bacterial meningitis.

  14. Spontaneous metastasis in matrix metalloproteinase 3-deficient mice

    DEFF Research Database (Denmark)

    Juncker-Jensen, Anna; Rømer, John; Pennington, Caroline J

    2009-01-01

    Matrix metalloproteinases (MMPs) have been linked to the metastatic potential of tumor cells due to their ability to degrade the extracellular matrix. MMP-3 (stromelysin-1) is upregulated in a wide variety of human tumors. We used the MMTV-PyMT breast cancer model to determine if MMP-3 is involved...

  15. Isolation and characterization of chicken bile matrix metalloproteinase

    Science.gov (United States)

    Avian bile is rich in matrix metalloproteinases (MMP), the enzymes that cleave extracellular matrix (ECM) proteins such as collagens and proteoglycans. Changes in bile MMP expression have been correlated with hepatic and gall bladder pathologies but the significance of their expression in normal, he...

  16. Altered neuronal architecture and plasticity in the visual cortex of adult MMP-3-deficient mice.

    Science.gov (United States)

    Aerts, Jeroen; Nys, Julie; Moons, Lieve; Hu, Tjing-Tjing; Arckens, Lutgarde

    2015-09-01

    Matrix metalloproteinases (MMPs) are Zn(2+)-dependent endopeptidases considered to be essential for normal brain development and neuroplasticity by modulating extracellular matrix proteins, receptors, adhesion molecules, growth factors and cytoskeletal proteins. Specifically, MMP-3 has recently been implicated in synaptic plasticity, hippocampus-dependent learning and neuronal development and migration in the cerebellum. However, the function(s) of this enzyme in the neocortex is understudied. Therefore, we explored the phenotypical characteristics of the neuronal architecture and the capacity for experience-dependent cortical plasticity in the visual cortex of adult MMP-3-deficient (MMP-3(-/-)) mice. Golgi-Cox stainings revealed a significant reduction in apical dendritic length and an increased number of apical obliques for layer V pyramidal neurons in the visual cortex of adult MMP-3(-/-) mice compared to wild-type (WT) animals. In addition, a significant upregulation of both phosphorylated and non-phosphorylated neurofilament protein (NF)-high, phosphorylated NF-medium, NF-low and α-internexin was detected in the visual cortex of MMP-3(-/-) mice. To assess the effect of MMP-3 deficiency on cortical plasticity, we monocularly enucleated adult MMP-3(-/-) mice and analyzed the reactivation of the contralateral visual cortex 7 weeks post-enucleation. In contrast to previous results in C57Bl/6J adult mice, activity remained confined to the binocular zone and did not expand into the monocular regions indicative for an aberrant open-eye potentiation. Permanent hypoactivity in the monocular cortex lateral and medial to V1 also indicated a lack of cross-modal plasticity. These observations demonstrate that genetic inactivation of MMP-3 has profound effects on the structural integrity and plasticity response of the visual cortex of adult mice.

  17. MMP-1/PAR-1 signal transduction axis and its prognostic impact in esophageal squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Hong-hua Peng

    2012-01-01

    Full Text Available The matrix metalloprotease-1 (MMP-1/protease-activated receptor-1 (PAR-1 signal transduction axis plays an important role in tumorigenesis. To explore the expression and prognostic value of MMP-1 and PAR-1 in esophageal squamous cell carcinoma (ESCC, we evaluated the expression of two proteins in resected specimens from 85 patients with ESCC by immunohistochemistry. Sixty-two (72.9% and 58 (68.2% tumors were MMP-1- and PAR-1-positive, respectively, while no significant staining was observed in normal esophageal squamous epithelium. MMP-1 and PAR-1 overexpression was significantly associated with tumor node metastasis (TNM stage and regional lymph node involvement. Patients with MMP-1- and PAR-1-positive tumors, respectively, had poorer disease-free survival (DFS than those with negative ESCC (P = 0.002 and 0.003, respectively. Univariate analysis showed a significant relationship between TNM stage [hazard ratio (HR = 2.836, 95% confidence interval (CI = 1.866-4.308], regional lymph node involvement (HR = 2.955, 95%CI = 1.713-5.068, MMP-1 expression (HR = 2.669, 95%CI = 1.229-6.127, and PAR-1 expression (HR = 1.762, 95%CI = 1.156-2.883 and DFS. Multivariate analysis including the above four parameters identified TNM stage (HR = 2.035, 95%CI = 1.167-3.681, MMP-1 expression (HR = 2.109, 95%CI = 1.293-3.279, and PAR-1 expression (HR = 1.967, 95%CI = 1.256-2.881 as independent and significant prognostic factors for DFS. Our data suggest for the first time that MMP-1 and PAR-1 were both overexpressed in ESCC and are novel predictors of poor patient prognosis after curative resection. The MMP-1/PAR-1 signal transduction axis might be a new therapeutic target for future therapies tailored against ESCC.

  18. MMP-1/PAR-1 signal transduction axis and its prognostic impact in esophageal squamous cell carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Hong-hua; Zhang, Xi; Cao, Pei-guo [Department of Oncology, the Third Xiangya Hospital, Central South University, Changsha, Hunan Province (China)

    2011-11-18

    The matrix metalloprotease-1 (MMP-1)/protease-activated receptor-1 (PAR-1) signal transduction axis plays an important role in tumorigenesis. To explore the expression and prognostic value of MMP-1 and PAR-1 in esophageal squamous cell carcinoma (ESCC), we evaluated the expression of two proteins in resected specimens from 85 patients with ESCC by immunohistochemistry. Sixty-two (72.9%) and 58 (68.2%) tumors were MMP-1- and PAR-1-positive, respectively, while no significant staining was observed in normal esophageal squamous epithelium. MMP-1 and PAR-1 overexpression was significantly associated with tumor node metastasis (TNM) stage and regional lymph node involvement. Patients with MMP-1- and PAR-1-positive tumors, respectively, had poorer disease-free survival (DFS) than those with negative ESCC (P = 0.002 and 0.003, respectively). Univariate analysis showed a significant relationship between TNM stage [hazard ratio (HR) = 2.836, 95% confidence interval (CI) = 1.866-4.308], regional lymph node involvement (HR = 2.955, 95%CI = 1.713-5.068), MMP-1 expression (HR = 2.669, 95%CI = 1.229-6.127), and PAR-1 expression (HR = 1.762, 95%CI = 1.156-2.883) and DFS. Multivariate analysis including the above four parameters identified TNM stage (HR = 2.035, 95%CI = 1.167-3.681), MMP-1 expression (HR = 2.109, 95%CI = 1.293-3.279), and PAR-1 expression (HR = 1.967, 95%CI = 1.256-2.881) as independent and significant prognostic factors for DFS. Our data suggest for the first time that MMP-1 and PAR-1 were both overexpressed in ESCC and are novel predictors of poor patient prognosis after curative resection. The MMP-1/PAR-1 signal transduction axis might be a new therapeutic target for future therapies tailored against ESCC.

  19. Epigenetic regulation of proMMP-1 expression in the HT1080 human fibrosarcoma cell line.

    Science.gov (United States)

    Poplineau, Mathilde; Dufer, Jean; Antonicelli, Frank; Trussardi-Regnier, Aurelie

    2011-06-01

    The matrix metalloproteinase (MMP) family members play an important role in various physiological and pathological processes. Although MMP-1 (collagenase-1) has been shown to be involved in tumor invasiveness, the regulation of its expression is still not fully elucidated and could implicate epigenetic mechanisms. The aim of this study was to analyze the effects of the Histone Deacetylase Inhibitor (HDI) trichostatin A (TSA) and the inhibitor of DNA methylation 5-aza-2'-deoxycytidine (5-azadC) on the proMMP-1 expression in the human HT1080 fibrosarcoma cell line. Real-time RT-PCR revealed that 5-azadC or 5-azadC + TSA but not TSA alone, despite global histone H4 hyperacetylation, increased proMMP-1 mRNA levels. This transcription activation was correlated with chromatin decondensation determined by nuclear texture image analysis technique. Western blot analysis of cell culture conditioned media revealed a significant increase in proMMP-1 secretion after 5-azadC or 5-azadC + TSA treatment compared to untreated cells. These results suggested that epigenetic mechanisms could be involved in proMMP-1 gene expression including chromatin supra-organization changes. Indeed, although the proMMP-1 gene promoter does not appear to contain CpG islands, its expression can be induced by the demethylating agent 5-azadC. Further experiments revealed that inhibition of protein neosynthesis by cycloheximide decreased 5-azadC-induced proMMP-1 mRNA, suggesting that epigenetically regulated intermediate molecules could be involved in proMMP-1 expression regulation in these cells.

  20. Lymph node micrometastasis and its correlation with MMP-2 expression in gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    Ze-Yu Wu; Jing-Hua Li; Wen-Hua Zhan; Yu-Long He

    2006-01-01

    AIM: To examine matrix metalloproteinase-2 (MMP-2)expression in gastric cancer tissues and to evaluate its relationship with lymph node micrometastasis.MATERIALS: The authors studied 850 lymph nodes resected from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenetomy using reverse transcription polymerase chain reaction (RT-PCR)assay in addition to H-E staining. MMP-2 expression of the tumor tissues was detected by immunohistochemical technique (EliVisionTM plus).RESULTS: MMP-2 expression was positive in 21 (70%)cases and negative in 9 (30%) cases. No significant correlations were found between MMP-2 expression and other variables such as age, gender, tumor location,tumor diameter, Lauren classification and lymphatic invasion. In contrast, MMP-2 expression correlated significantly with depth of tumor infiltration (P =0.022), lymph node metastasis (P = 0.030) and tumor differentiation (P = 0.043). Lymph node micrometastases were detected in 77 (12.5%) lymph nodes of 14 (46.7%)gastric carcinoma patients. MMP-2 expression was positive in 12 (85.7%) of the 14 patients with lymph node micrometastasis, and in 9 (56.3%) of the 16patients without lymph node micrometastasis (P = 0.118).CONCLUSIONS: Our results demonstrate that MMP-2 expression has significant correlation with tumor invasion, tumor differentiation and lymph node metastases. MMP-2 expression may be an important biological characteristics and significant prognostic parameter of gastric carcinoma. We also conclude that MMP-2 may participate in the development of lymph node micrometastasis of gastric carcinoma. Further investigations are needed to draw a conclusion.

  1. Serum level of MMP-2, MMP-9 and Ox-LDL in Alzheimer's disease with hyperlipoidemia

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective:To investigate serum levels of MMP-2,MMP-9, oxidized low density lipoprotein (ox-LDL) in Alzheimer's disease (AD) patients and study the possible pathway and mechanism of AD with abnormal lipid metabolism. Methods: Subjects in this study were divided into 4 groups: normal lipid group without AD (N), hyperlipoidemia group without AD (H), normal group with AD (A), hyperlipoidemia group with AD (AH). There were 15 individuals in each group. MMP-2, MMP-9, ox-LDL was measured by enzyme linked immunosorbent assay (ELISA). Serum lipids levels were measured by biochemical methods. Results: The serum levels of MMP-2, MMP-9, ox-LDL were significantly higher in H, A and AH groups than those in N group. Those of ox LDL in H, AH groups was higher than that of in A group. The serum level of MMP-2, MMP-9 in AH groups were higher than that of in H group. The score of mini-mental state examination (MMSE) in A and AD groups was negatively correlated with the serum level of ox-LDL. Relationship between the score of MMSE and the serum level of ox-LDL in AD groups and non-AD groups had statistical significance. Conclusion: MMP-2, MMP-9, ox-LDL and abnormal lipid metabolism may participate in pathogenesis of AD, in which abnormal lipid metabolism induces expressions of MMP-2,MMP-9 and ox-LDL. Oxidative stress and blood-brain barrier disruption might accelerate the process of AD.

  2. The active form of MMP-3 is a marker of synovial inflammation and cartilage turnover in inflammatory joint diseases

    DEFF Research Database (Denmark)

    Sun, Shu; Bay-Jensen, Anne-Christine; Karsdal, Morten A

    2014-01-01

    ) and erythrocyte sedimentation rate (ESR). In addition, in patients receiving anti-TNF-α treatment, the serum level of act-MMP-3 was significantly reduced compared to baseline level reflecting the anti-inflammatory effects of the treatment. CONCLUSION: We have successfully developed an assay measuring act-MMP-3......BACKGROUND: Matrix metalloproteinase-3 (MMP-3) plays an important role in the pathology of rheumatoid arthritis (RA) and ankylosing spondylitis (AS). Measurement of active MMP-3 in clinical samples could provide information about progression of rheumatoid diseases, and potentially response...... and after anti-TNF-α treatment were evaluated. RESULTS: A highly specific and technically robust ELISA detecting act-MMP-3 in serum was developed. The lower limit of detection was 33.7 pg/mL. The dilution and spiking recovery of human serum was within 100 ± 20%. The average intra- and inter-assay variations...

  3. A PKC-dependent recruitment of MMP-2 controls semaphorin-3A growth-promoting effect in cortical dendrites.

    Directory of Open Access Journals (Sweden)

    Bertrand Gonthier

    Full Text Available There is increasing evidence for a crucial role of proteases and metalloproteinases during axon growth and guidance. In this context, we recently described a functional link between the chemoattractive Sema3C and Matrix metalloproteinase 3 (MMP3. Here, we provide data demonstrating the involvement of MMP-2 to trigger the growth-promoting effect of Sema3A in cortical dendrites. The in situ analysis of MMP-2 expression and activity is consistent with a functional growth assay demonstrating in vitro that the pharmacological inhibition of MMP-2 reduces the growth of cortical dendrites in response to Sema3A. Hence, our results suggest that the selective recruitment and activation of MMP-2 in response to Sema3A requires a PKC alpha dependent mechanism. Altogether, we provide a second set of data supporting MMPs as effectors of the growth-promoting effects of semaphorins, and we identify the potential signalling pathway involved.

  4. Inhibition of MMP-9 by green tea catechins and prediction of their interaction by molecular docking analysis.

    Science.gov (United States)

    Sarkar, Jaganmay; Nandy, Suman Kumar; Chowdhury, Animesh; Chakraborti, Tapati; Chakraborti, Sajal

    2016-12-01

    Green tea polyphenolic catechins have been shown to prevent various types of diseases such as pulmonary hypertension (PAH), cancer and cardiac and neurological disorders. Matrix metalloproteinases (MMPs) play an important role in the development of PAH. The present study demonstrated that among the four green tea catechins (EGCG, ECG, EC and EGC), EGCG and ECG inhibit pro-/active MMP-9 activities in pulmonary artery smooth muscle cell culture supernatant. Based on the above, we investigated the interactions of pro-/active MMP-9 with the green tea catechins by computational methods. In silico molecular docking analysis revealed a strong interaction between pro-/active MMP-9 and EGCG/ECG, and galloyl group appears to be responsible for this enhanced interaction. The molecular docking studies corroborate our experimental observation that EGCG and ECG are mainly active in preventing both the proMMP-9 and MMP-9 activities.

  5. MMP-8 C-799T and MMP-8 C+17G polymorphisms in mild and severe preeclampsia: Association between MMP-8 C-799T with susceptibility to severe preeclampsia.

    Science.gov (United States)

    Rahimi, Ziba; Zangeneh, Maryam; Rezaeyan, Arezoo; Shakiba, Ebrahim; Rahimi, Zohreh

    2017-07-26

    The aim of present study was to determine the role of matrix metalloproteinase-8 (MMP-8) C-799T (rs11225395) and C+17 (rs2155052) polymorphisms in susceptibility to preeclampsia. In a case-control study, 256 pregnant women including 152 women with preeclampsia (86 women with mild preeclampsia and 66 women with severe preeclampsia) and 104 women with normal pregnancy from Western Iran with Kurdish ethnic background were investigated for MMP-8 C-799T and C + 17G polymorphisms using polymerase chain reaction-restriction fragment length polymorphism method. Comparing the MMP-8 TT genotype with the combined genotype of CC+CT (recessive model) indicated a significantly higher frequency of the MMP-8 TT genotype (47%) in severe preeclamptic patients than that in healthy pregnant women (30.8%) that was associated with 1.99-fold increased risk of severe preeclampsia (95% CI = 1.05-3.77, p = 0.034). The frequency of MMP-8 G allele was 27.3% in all preeclamptic patients compared to 30.2% in controls (p = 0.56). Also, no significant difference was detected comparing the frequency of G allele in mild (26.6%, p = 0.46) and severe preeclamptic patients (28.4%, p = 0.75) with controls (30.2%). Our study demonstrated that the MMP-8 C-799T is associated with the risk of developing severe preeclampsia during pregnancy. However, the MMP-8 C + 17G polymorphism might not be a risk factor for susceptibility to preeclampsia.

  6. MAPKs are essential upstream signaling pathways in proteolytic cartilage degradation--divergence in pathways leading to aggrecanase and MMP-mediated articular cartilage degradation

    DEFF Research Database (Denmark)

    Sondergaard, B-C; Schultz, N; Madsen, S H;

    2010-01-01

    Matrix metalloproteinases (MMPs) and aggrecanases are essential players in cartilage degradation. However, the signaling pathways that results in MMP and/or aggrecanase synthesis and activation are not well understood. We investigated the molecular events leading to MMP- and aggrecanase-mediated ...

  7. Effects of MMP inhibitors incorporated within dental adhesives.

    Science.gov (United States)

    Almahdy, A; Koller, G; Sauro, S; Bartsch, J W; Sherriff, M; Watson, T F; Banerjee, A

    2012-06-01

    Matrix metalloproteinase (MMP) inhibition has been shown to reduce adhesive bond degradation when applied as a pre-conditioner, adding to clinical steps in the placement of adhesives, but their incorporation within dental adhesives has not been fully explored. This study examined the effect of including 2 MMP inhibitors (BB94 and GM6001) within the primers of 3 commercially available adhesives. Fluorometric assay and zymography showed that adhesives with MMP inhibitors had high affinity toward both synthetic fluorogenic FRET peptides (95%) and dentin powder substrates, respectively. The immediate microtensile bond strength was enhanced for 2 types of adhesives following the addition of both inhibitors. However, no changes were detected between the control and the inhibitor groups following 3-month storage. The modified two-step etch-and-rinse and single-step systems showed less Rhodamine B penetration to the "hybrid layer" and to the "adhesive", respectively. The incorporation of BB94 and GM6001 within the primers resulted in the inhibition of dentin MMPs with improved initial bond strength and enhanced sealing ability.

  8. Stimulation of MMP-11 (stromelysin-3 expression in mouse fibroblasts by cytokines, collagen and co-culture with human breast cancer cell lines

    Directory of Open Access Journals (Sweden)

    Matthaei Klaus I

    2004-07-01

    Full Text Available Abstract Background Matrix metalloproteinases (MMPs are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling. MT1-MMP (MMP-14 and stromelysin-3 (MMP-11 are two members of the MMP family of proteolytic enzymes that have been specifically implicated in breast cancer progression. Expressed in stromal fibroblasts adjacent to epithelial tumour cells, the mechanism of MT1-MMP and MMP-11 induction remains unknown. Methods To investigate possible mechanisms of induction, we examined the effects of a number of plausible regulatory agents and treatments that may physiologically influence MMP expression during tumour progression. Thus NIH3T3 and primary mouse embryonic fibroblasts (MEFs were: a treated with the cytokines IL-1β, IL-2, IL-6, IL-8 and TGF-β for 3, 6, 12, 24, and 48 hours; b grown on collagens I, IV and V; c treated with fibronectin, con-A and matrigel; and d co-cultured with a range of HBC (human breast cancer cell lines of varied invasive and metastatic potential. Results Competitive quantitative RT-PCR indicated that MMP-11 expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-1β, IL-2, TGF-β, fibronectin and collagen V. No other substantial changes in expression of MMP-11 or MT1-MMP in either tested fibroblast culture, under any treatment conditions, were observed. Conclusion We have demonstrated significant MMP-11 stimulation in mouse fibroblasts using cytokines, matrix constituents and HBC cell lines, and also some inhibition of MT1-MMP. Our data suggest that the regulation of these genes in the complex stromal-epithelial interactions that occur in human breast carcinoma, is influenced by several mechanisms.

  9. Variants of the Matrix Metalloproteinase-2 but not the Matrix Metalloproteinase-9 genes significantly influence functional outcome after stroke

    Directory of Open Access Journals (Sweden)

    Sobral João

    2010-03-01

    Full Text Available Abstract Background Multiple lines of evidence suggest that genetic factors contribute to stroke recovery. The matrix metalloproteinases -2 (MMP-2 and -9 (MMP-9 are modulators of extracellular matrix components, with important regulatory functions in the Central Nervous System (CNS. Shortly after stroke, MMP-2 and MMP-9 have mainly damaging effects for brain tissue. However, MMPs also have a beneficial activity in angiogenesis and neurovascular remodelling during the delayed neuroinflammatory response phase, thus possibly contributing to stroke functional recovery. Methods In the present study, the role of MMP-2 and MMP-9 genetic variants in stroke recovery was investigated in 546 stroke patients. Functional outcome was assessed three months after a stroke episode using the modified Rankin Scale (mRS, and patients were classified in two groups: good recovery (mRS ≤ 1 or poor recovery (mRS>1. Haplotype tagging single nucleotide polymorphisms (SNPs in the MMP-2 (N = 21 and MMP-9 (N = 4 genes were genotyped and tested for association with stroke outcome, adjusting for significant non-genetic clinical variables. Results Six SNPs in the MMP-2 gene were significantly associated with stroke outcome (0.0018P P MMP-9 gene. Conclusions The results presented strongly indicate that MMP-2 genetic variants are an important mediator of functional outcome after stroke.

  10. Effect of gene CTGF transfection on the expression of MMP-2 and MMP-9 and proliferation in human cervical cancer cells%CTGF基因转染对宫颈癌细胞MMP-2、MMP-9表达及细胞增殖的影响

    Institute of Scientific and Technical Information of China (English)

    肖蔚; 焦霞; 钱华; 崔永安; 林梅; 王薇; 周彤敏; 窦荣荣; 于鸿

    2012-01-01

    Objective: To investigate the effect of connective tissue growth factor ( CTGF ) on the proliferation in human cervical cancer cell line Hela, with a focus on the expression of matrix metalloproteinase-2( MMP-2 ) and matrix metalloproteinase-9( MMP-9 ), and explore the underlying mechanism for the role of CTGF in the development of cervical cancer. Methods: pcDNA3. 0-CTGF and pcDNA3.0 were transfected into Hela cells through lipofectamine and positive clones which were screened by G418. Fluorescence quan-titive polymerase chain reaction( FQ-PCR ) and Western blot were employed to identify mRNA and protein expression of CTGF in Hela cells, respectively. The expression of MMP-2 and MMP-9 in positive clones was detected by FQ-PCR and Western blot. Cell viability was assessed by dimethylthiazoldiphenyl-tetrazolium-bromide ( MTT ) method. Results: Positive clone C-16 with CTGF over-expression were successfully established. Compared with non-transfected control group, the expression of MMP-2 and MMP-9 in C-16 were increased significantly,and the proliferation level of C-16 was increased significantly. Conclusion: CTGF transfection could effectively enhance the expression of MMP-2 and MMP-9 and the proliferation in Hela cells which suggested a potential role for CTGF in gene therapy of cervical cancer.%目的:研究结缔组织生长因子(connective tissue growth factor,CTGF)基因对人宫颈癌Hela细胞基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达及细胞增殖的影响,探讨CTGF在宫颈癌侵袭和转移中的作用机制.方法:经脂质体介导将含有CTGF重组表达质粒转染人宫颈癌Hela细胞株,用G418筛选阳性细胞克隆及实时荧光定量PCR、蛋白质印迹鉴定;采用实时荧光定量PCR和蛋白质印迹法检测阳性克隆细胞MMP-2及MMP-9的表达;噻唑盐(MTT)比色法检测阳性细胞克隆的增殖活性.结果:成功建立稳定高表达CTGF的阳性Hela细胞克隆,证实其MMP-2、MMP-9表达及细

  11. Activation of Proteinkinase ERK Mediates Induction of Macrophage MMP-12 by OxLDL

    Institute of Scientific and Technical Information of China (English)

    He Chun-yan; Zhou Xin; Li Xiao-ming; Yu Hong; Hong Jia-ling

    2004-01-01

    The present study was undertaken to investigate the effect of oxidized low density lipoprotein (oxLDL) on the expression of macrophage matrix metalloproteinase-12 (MMP-12), and the possible mechanisms. Activation of extracellular signal-regulated kinase 1/2 (ERK1/2) was detected by Western blot analysis. Enzymatic activity of MMP-12 was determined by β-casein zymogra-phy. RT-PCR analysis was used to measure the mRNA expression level of MMP-12. OxLDL-stimulated macrophages produced increased casein-degrading activities and oxLDL also significantly increased the mRNA level of MMP-12 in a dose-dependent manner. OxLDL stimulated the phosphorylation of ERK1/2 in macrophages. The use of the specific inhibitor indicated that the ERK1/2 signaling pathway was required for the induction of MMP-12. These data demonstrated that oxLDL induced MMP-12 expression in macrophages through an ERK1/2-dependent pathway.

  12. Angiogenin contributes to bladder cancer tumorigenesis by DNMT3b-mediated MMP2 activation.

    Science.gov (United States)

    Peres, Rafael; Furuya, Hideki; Pagano, Ian; Shimizu, Yoshiko; Hokutan, Kanani; Rosser, Charles J

    2016-07-12

    Epigenetic-mediated gene activation/silencing plays a crucial role in human tumorigenesis. Eliciting the underlying mechanism behind certain epigenetic changes is essential for understanding tumor biology. Previous studies in human cancers revealed an unrecognized interplay between Angiogenin (ANG) and matrix metalloproteinase-2 (MMP2) leading to pronounced tumorigenesis. Here we provide multiple lines of evidence further indicating ANG oncogenic potential. ANG expression resulted in the hypomethylated state of the MMP2 gene, which led to increased gene expression of MMP2. More than that, our global DNA methylation microarray analysis showed that gene manipulation of ANG affected a variety of pathways, such as cell migration, angiogenesis and specifically, tumor suppressor genes. Mechanistically, ANG negatively regulated DNA methyltransferase 3b (DNMT3b) enzymatic activity by down-regulating its expression and inhibiting its recruitment to the MMP2 promoter. Consistent with this, ANG-MMP2 overexpression and DNMT3b underexpression correlated with reduction in disease free survival of human bladder cancer patients. Together, the results continue to establish ANG as an oncoprotein and further reveal that ANG contributes to oncogenesis by the activation of MMP2 through modulation of DNMT3b functions.

  13. New insight into the role of MMP14 in metabolic balance

    Directory of Open Access Journals (Sweden)

    Hidetoshi Mori

    2016-07-01

    Full Text Available Membrane-anchored matrix metalloproteinase 14 (MMP14 is involved broadly in organ development through both its proteolytic and signal-transducing functions. Knockout of Mmp14 (KO in mice results in a dramatic reduction of body size and wasting followed by premature death, the mechanism of which is poorly understood. Since the mammary gland develops after birth and is thus dependent for its functional progression on systemic and local cues, we chose it as an organ model for understanding why KO mice fail to thrive. A global analysis of the mammary glands’ proteome in the wild type (WT and KO mice provided insight into an unexpected role of MMP14 in maintaining metabolism and homeostasis. We performed mass spectrometry and quantitative proteomics to determine the protein signatures of mammary glands from 7 to 11 days old WT and KO mice and found that KO rudiments had a significantly higher level of rate-limiting enzymes involved in catabolic pathways. Glycogen and lipid levels in KO rudiments were reduced, and the circulating levels of triglycerides and glucose were lower. Analysis of the ultrastructure of mammary glands imaged by electron microscopy revealed a significant increase in autophagy signatures in KO mice. Finally, Mmp14 silenced mammary epithelial cells displayed enhanced autophagy. Applied to a systemic level, these findings indicate that MMP14 is a crucial regulator of tissue homeostasis. If operative on a systemic level, these findings could explain how Mmp14KO litter fail to thrive due to disorder in metabolism.

  14. Involvement of CD147 in overexpression of MMP-2 and MMP-9 and enhancement of invasive potential of PMA-differentiated THP-1

    Directory of Open Access Journals (Sweden)

    Tang Hao

    2005-05-01

    Full Text Available Abstract Background During infection and inflammation, circulating blood monocytes migrate from the intravascular compartments to the extravascular compartments, where they mature into tissue macrophages. The maturation process prepares the cells to actively participate in the inflammatory and immune responses, and many factors have been reported to be involved in the process. We found in our study that CD147 played a very important role in this process. Results By using PMA-differentiated human monocyte cells line THP-1, we found that CD147 mediated matrix metalloproteinases (MMPs expression of the leukemic THP-1 cells and thus enhanced the invasiveness of THP-1 cells. After 24 hours of PMA-induced monocyte differentiation, the mean fluorescence intensity of CD147 in differentiated THP-1 cells (289.61 ± 31.63 was higher than that of the undifferentiated THP-1 cells (205.1 ± 19.25. There was a significant increase of the levels of proMMP-2, proMMP-9 and their activated forms in the differentiated THP-1 cells. Invasion assays using reconstituted basement membrane showed a good correlation between the invasiveness of THP-1 cells and the production of MMP-2 and MMP-9. The difference in the MMPs expression and the invasive ability was significantly blocked by HAb18G/CD147 antagonistic peptide AP-9. The inhibitory rate of the secretion of proMMP-9 in the undifferentiated THP-1 cells was 45.07%. The inhibitory rate of the secretion of proMMP-9, the activated MMP-9 and proMMP-2 in the differentiated THP-1 cells was 52.90%, 53.79% and 47.80%, respectively. The inhibitory rate of invasive potential in the undifferentiated cells and the differentiated TH