DOCSLIB.ORG

ADAM17 Targets MMP-2 and MMP-9 Via EGFR-MEK-ERK Pathway Activation to Promote Prostate Cancer Cell Invasion

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
ADAM17 Targets MMP-2 and MMP-9 Via EGFR-MEK-ERK Pathway Activation to Promote Prostate Cancer Cell Invasion 1714 INTERNATIONAL JOURNAL OF ONCOLOGY 40: 1714-1724, 2012 ADAM17 targets MMP-2 and MMP-9 via EGFR-MEK-ERK pathway activation to promote prostate cancer cell invasion LI-JIE XIAO1,2*, PING LIN1*, FENG LIN1*, XIN LIU1, WEI QIN1, HAI-FENG ZOU1, LIANG GUO1, WEI LIU1, SHU-JUAN WANG1 and XIAO-GUANG YU1 1Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Harbin Medical University, 194 Xuefu Road, Harbin 150081, Heilongjiang; 2College of Life Science and Technology, Heilongjiang Bayi Agricultural University, 2 Xinyang Road, Daqing 163319, Heilongjiang, P.R. China Received September 8, 2011; Accepted November 18, 2011 DOI: 10.3892/ijo.2011.1320 Abstract. ADAM17, also known as tumor necrosis factor-α released and down-regulation of MMP-2, MMP-9. However, converting enzyme (TACE), is involved in proteolytic ectodo- these effects could be reversed by simultaneous addition of main shedding of cell surface molecules and cytokines. TGF-α. These data demonstrated that ADAM17 contributes to Although aberrant expression of ADAM17 has been shown androgen-independent prostate cancer cell invasion by shed- in various malignancies, the function of ADAM17 in prostate ding of EGFR ligand TGF-α, which subsequently activates the cancer has not been clarified. In the present study, we sought EGFR-MEK-ERK signaling pathway, leading finally to over- to elucidate whether ADAM17 contributes to prostate cancer expression of MMP-2 and MMP-9. This study suggests that the cell invasion, as well as the mechanism involved in the process. ADAM17 expression level may be a new predictive biomarker The expression pattern of ADAM17 was investigated in human of invasion and metastasis of prostate cancer, and ADAM17 prostate cancer cells. The results showed that ADAM17 could provide a target for treating metastatic PCa. expression levels are correlated with the invasive ability of androgen-independent prostate cancer cell lines. Further, Introduction ADAM17 was overexpressed in cells showing high invasion characteristics, activation of the EGFR-MEK-ERK pathway, Prostate cancer (PCa) is one of the most common malignancies up-regulation of MMP-2, MMP-9, and an increased TGF-α among Western males with an occurrence of approximately release into the supernatant. However, AG1478, PD98059 and 192,280 new cases and 27,360 deaths in the US, in 2009 antibody against TGF-α deactivating the EGFR-MEK-ERK (1). The growth and maintenance of cancerous cells in the signaling pathway, abolished up-regulation of MMP-2, MMP-9 early stages of prostate cancer is androgen-dependent and and prevented cell invasion. In addition, cells with knockdown therapy often involves androgen deprivation (2). In the later of ADAM17 by siRNA exhibited low invasive ability, deacti- stages therapeutic alternatives are not available, as the tumor vated EGFR-MEK-ERK signaling pathway, reduced TGF-α progresses from androgen-dependence to -independence. The lack of effective therapies for advanced PCa is related to a large extent to poor understanding of the molecular mechanisms underlying the progression of disease toward invasion and Correspondence to: Professor Xiao-Guang Yu, Department of metastasis (3). Therefore, the identification of new predictive Biochemistry and Molecular Biology, College of Basic Medical biomarkers, especially those that are indicative of invasiveness Science, Harbin Medical University, 194 Xuefu Road, Harbin of the disease, which could serve as targets for establishing 150081, Heilongjiang, P.R. China effectiveness of therapeutic and chemopreventive interventions, E-mail: [email protected] will improve clinical management of PCa (4). * The ADAMs (a disintegrin and metalloproteinase) are Contributed equally members of the metzincin superfamily of Zn-dependent matrix metalloproteinases. They are transmembrane and secreted Abbreviations: ADAM17, a disintegrin and metalloprotease 17; ADAMs, a disintegrin and metalloproteinase; TACE, tumor necrosis proteins with important roles in diverse biological functions, factor-α converting enzyme; MMP-2 and MMP-9, matrix metallo- such as fertilization, adhesion, migration, proteolysis and proteinase-2 and -9; TGF-α, transforming growth factor-α; EGFR, signaling (5,6). Recently, specific ADAMs were implicated in a epidermal growth factor receptor; Akt, mitogenactivated protein broad range of diseases, including Alzheimer's disease, asthma, kinase; ERK, extracellular signal-regulated protein kinase; ECM, diabetes, rheumatoid arthritis, atherosclerosis, kidney fibrosis, extracellular matrix and cancer (7,8). To date, 21 functional ADAMs have been described in humans and 40 in different organisms (9). Out of Key words: a disintegrin and metalloprotease 17, transforming this long list of ADAMs, the best characterized ADAM family growth factor-α, prostatic adenocarcinomas, matrix metallopro- member is ADAM17. ADAM17, also known as tumor necrosis teinase-2 and -9 factor-α converting enzyme (TACE) was originally described XIAO et al: ADAM17 TARGETS MMP-2 AND MMP-9 TO PROMOTE PROSTATE CANCER 1715 as being responsible for the proteolytic cleavage of the soluble are as follows: ADAM17-siRNA1, 5'-AUGAGUUGUAACCAG form of tumor necrosis factor-α (TNF-α), which is located on GUCAGCUUCC-3'; ADAM17-siRNA2, 5'-AUACAUGACAUA the cell surface (10,11). However, subsequent studies showed UUUCCCUCCCUGG-3'. DU145 cells grown to 40-60% conflu- that ADAM17 also sheds a variety of important cell surface ency were transfected with ADAM17 siRNA or a negative molecules, including growth factors (12), adhesion molecules control siRNA using Lipofectamine 2000 (Invitrogen) according (13,14) and chemokines (15,16). Any of these play key roles in to manufacturer's protocol. The transfected cells were harvested a variety of diseases, including cancer. Aberrant expression of at 48 or 72 h for mRNA isolation and protein extraction, ADAM17 has been shown in various malignancies (17-19). respectively. However, reports on the expression and function of Transfections of PC-3-ADAM17 cells with MMP-2-siRNA ADAM17 in prostate cancer are limited. ADAM17 mRNA or MMP-9-siRNA were performed as described above. expression was found in both androgen-dependent and Nucleotide synthetic duplexes of MMP-2-siRNA and MMP-9- androgen-independent prostate cancer cell lines (2). Subsequent siRNA were obtained from Invitrogen. Sequences are as study showed ADAM17 expression in non-cancerous (BPH) follows: MMP-2-siRNA, 5'-AGUAGAUCCAGUAUUCAUUC prostatic tissues was lower than that in tumor samples (20). CCUGC-3'; MMP-9-siRNA, 5'-AAUACAGCUGGUUCCCAA Although the expression pattern for ADAM17 was determined UCUCCGC-3'. in human prostatic adenocarcinomas, no functional assays were performed in these studies, where the expression has been Semi-quantitative real-time PCR. Total RNA from control investigated in a very limited way. Nevertheless, these studies treated cells was extracted using TRIzol reagent (Invitrogen). raise the possibility of a functional role of ADAM17 in prostatic Then, 2 µg RNA was converted to cDNA with Superscript II carcinomas. Therefore, the goal of this study was to investigate reverse transcriptase (Invitrogen). The cDNA product was mixed whether ADAM17 contributes to prostate cancer cell invasion with SYBR® Green PCR Master Mix (Applied Biosystems) and its mechanisms. and specific primers of ADAM17 or β-actin in the subsequent PCR using ABI7500 real-time PCR system (Applied Biosys- Materials and methods tems). Primers used in this study were as follows: β-actin, sense: 5'-TACCTCATGAAGATCCTCACC-3'; antisense: 5'-TTTCG Materials. Primary antibodies against Akt, p-Akt, and p-EGFR TGGATGCCACAGGAC-3'; ADAM 17, sense: 5'-AGCAACAG were purchased from Cell Signaling Technology; antibodies CATCCACCTAAT-3'; antisense: 5'-CAGGAGTGTCTGGAA against ERK, p-ERK, MMP-2, MMP-9 and GAPDH were from TTGTC-3'. Annealing temperatures was 55˚C. Amplification Santa Cruz Biotechnology; ADAM17 antibody was obtained cycles were 30 for ADAM17 and 23 for β-actin. Each reaction from Abcam and EGFR antibody from SAB. HRP-labeled anti- was done in triplicate to minimize experimental variation and mouse or rabbit antibodies were from Santa Cruz Biotechnology. transcript levels of β-actin were monitored as an internal control. Geneticin and culture media were from Gibco (Grand Island, NY, USA) and Hyclone (Logan, UT), respectively. TGF-α and Western blot analysis. Cells treated with different compounds EGF were from R&D Systems (Minneapolis, MN) and AG1478, were harvested at indicated time, washed, and lysed in ice-cold LY294002, PD98059 were purchased from Cell Signaling lysis buffer (Beyotime Institute of Biotechnology, P.R. China) Technology (Danvers, MA, USA). All the primers used in this for 10 min on ice. The lysate was centrifuged at 12,000 g at research were synthesized by Shanghai Sangon Biotechnology 4˚C for 5 min. The protein concentration of extracts was deter- Co., Ltd. mined with BCA reagent (Beyotime Institute of Biotechnology, P.R. China). Equal amounts of protein (40 µg/lane) were Cell culture. Human prostate cancer cell line DU145 and PC-3 loaded, separated by 12% SDS-PAGE and transferred onto were purchased from American Type Culture Collection, and nitrocellulose membrane. After blocked with 5% non-fat milk, PC-3M, PC-3M-1E8 and PC-3M-2B4 were obtained from the the membranes were incubated with different primary antibody Peking University Health Science Center (Beijing, P.R. China). at 4˚C overnight, and then HRP-labeled secondary antibodies. Cells were maintained
Load more

Recommended publications
  • MATRIX METALLOPROTEINASE REQUIREMENTS for NEUROMUSCULAR SYNAPTOGENESIS by Mary Lynn Dear Dissertation Submitted to the Faculty O
    MATRIX METALLOPROTEINASE REQUIREMENTS FOR NEUROMUSCULAR SYNAPTOGENESIS By Mary Lynn Dear Dissertation Submitted to the Faculty of the Graduate School of Vanderbilt University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Biological Sciences January 31, 2018 Nashville, Tennessee Approved: Todd Graham, Ph.D. Kendal Broadie, Ph.D. Katherine Friedman, Ph.D. Barbara Fingleton, Ph.D. Jared Nordman, Ph.D. Copyright © 2018 by Mary Lynn Dear All Rights Reserved ACKNOWLEDGEMENTS I would like to acknowledge my advisor, Kendal Broadie, and the entire Broadie laboratory, both past and present, for their endless support, engaging conversations, thoughtful suggestions and constant encouragement. I would also like to thank my committee members both past and present for playing such a pivotal role in my graduate career and growth as a scientist. I thank the Department of Biological Sciences for fostering my graduate education. I thank the entire protease community for their continued support, helpful suggestions and collaborative efforts that helped my project move forward. I would like to acknowledge Dr. Andrea Page-McCaw and her entire laboratory for helpful suggestions, being engaged in my studies and providing many tools that were invaluable to my project. I thank my parents, Leisa Justus and Raymond Dear, my brother Jake Dear and my best friend Jenna Kaufman. Words cannot describe how grateful I am for your support and encouragement. Most importantly, I would like to thank my husband, Jeffrey Thomas, for always believing in me, your unwavering support and for helping me endure all the ups and downs that come with graduate school.
    [Show full text]
  • What Are the Roles of Metalloproteinases in Cartilage and Bone Damage? G Murphy, M H Lee
    iv44 Ann Rheum Dis: first published as 10.1136/ard.2005.042465 on 20 October 2005. Downloaded from REPORT What are the roles of metalloproteinases in cartilage and bone damage? G Murphy, M H Lee ............................................................................................................................... Ann Rheum Dis 2005;64:iv44–iv47. doi: 10.1136/ard.2005.042465 enzyme moiety into an upper and a lower subdomain. A A role for metalloproteinases in the pathological destruction common five stranded beta-sheet and two alpha-helices are in diseases such as rheumatoid arthritis and osteoarthritis, always found in the upper subdomain with a further C- and the irreversible nature of the ensuing cartilage and bone terminal helix in the lower subdomain. The catalytic sites of damage, have been the focus of much investigation for the metalloproteinases, especially the MMPs, have been several decades. This has led to the development of broad targeted for the development of low molecular weight spectrum metalloproteinase inhibitors as potential therapeu- synthetic inhibitors with a zinc chelating moiety. Inhibitors tics. More recently it has been appreciated that several able to fully differentiate between individual enzymes have families of zinc dependent proteinases play significant and not been identified thus far, although a reasonable level of varied roles in the biology of the resident cells in these tissues, discrimination is now being achieved in some cases.7 Each orchestrating development, remodelling, and subsequent family does, however, have other unique domains with pathological processes. They also play key roles in the numerous roles, including the determination of physiological activity of inflammatory cells. The task of elucidating the substrate specificity, ECM, or cell surface localisation (fig 1).
    [Show full text]
  • Matrix Metalloproteinases in Angiogenesis: a Moving Target for Therapeutic Intervention
    Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention William G. Stetler-Stevenson J Clin Invest. 1999;103(9):1237-1241. https://doi.org/10.1172/JCI6870. Perspective Angiogenesis is the process in which new vessels emerge from existing endothelial lined vessels. This is distinct from the process of vasculogenesis in that the endothelial cells arise by proliferation from existing vessels rather than differentiating from stem cells. Angiogenesis is an invasive process that requires proteolysis of the extracellular matrix and, proliferation and migration of endothelial cells, as well as synthesis of new matrix components. During embryonic development, both vasculogenesis and angiogenesis contribute to formation of the circulatory system. In the adult, with the single exception of the reproductive cycle in women, angiogenesis is initiated only in response to a pathologic condition, such as inflammation or hypoxia. The angiogenic response is critical for progression of wound healing and rheumatoid arthritis. Angiogenesis is also a prerequisite for tumor growth and metastasis formation. Therefore, understanding the cellular events involved in angiogenesis and the molecular regulation of these events has enormous clinical implications. This understanding is providing novel therapeutic targets for the treatment of a variety of diseases, including cancer. Whatever the pathologic condition, an initiating stimulus results in the formation of a migrating solid column of endothelial cells called the vascular sprout. The advancing front of this endothelial cell column presumably focuses proteolytic activity to create a defect in the extracellular matrix, through which the advancing and proliferating column of endothelial […] Find the latest version: https://jci.me/6870/pdf Matrix metalloproteinases in angiogenesis: Perspective a moving target for therapeutic intervention SERIES Topics in angiogenesis David A.
    [Show full text]
  • Gent Forms of Metalloproteinases in Hydra
    Cell Research (2002); 12(3-4):163-176 http://www.cell-research.com REVIEW Structure, expression, and developmental function of early diver- gent forms of metalloproteinases in Hydra 1 2 3 4 MICHAEL P SARRAS JR , LI YAN , ALEXEY LEONTOVICH , JIN SONG ZHANG 1 Department of Anatomy and Cell Biology University of Kansas Medical Center Kansas City, Kansas 66160- 7400, USA 2 Centocor, Malvern, PA 19355, USA 3 Department of Experimental Pathology, Mayo Clinic, Rochester, MN 55904, USA 4 Pharmaceutical Chemistry, University of Kansas, Lawrence, KS 66047, USA ABSTRACT Metalloproteinases have a critical role in a broad spectrum of cellular processes ranging from the breakdown of extracellular matrix to the processing of signal transduction-related proteins. These hydro- lytic functions underlie a variety of mechanisms related to developmental processes as well as disease states. Structural analysis of metalloproteinases from both invertebrate and vertebrate species indicates that these enzymes are highly conserved and arose early during metazoan evolution. In this regard, studies from various laboratories have reported that a number of classes of metalloproteinases are found in hydra, a member of Cnidaria, the second oldest of existing animal phyla. These studies demonstrate that the hydra genome contains at least three classes of metalloproteinases to include members of the 1) astacin class, 2) matrix metalloproteinase class, and 3) neprilysin class. Functional studies indicate that these metalloproteinases play diverse and important roles in hydra morphogenesis and cell differentiation as well as specialized functions in adult polyps. This article will review the structure, expression, and function of these metalloproteinases in hydra. Key words: Hydra, metalloproteinases, development, astacin, matrix metalloproteinases, endothelin.
    [Show full text]
  • Functional and Structural Insights Into Astacin Metallopeptidases
    Biol. Chem., Vol. 393, pp. 1027–1041, October 2012 • Copyright © by Walter de Gruyter • Berlin • Boston. DOI 10.1515/hsz-2012-0149 Review Functional and structural insights into astacin metallopeptidases F. Xavier Gomis-R ü th 1, *, Sergio Trillo-Muyo 1 Keywords: bone morphogenetic protein; catalytic domain; and Walter St ö cker 2, * meprin; metzincin; tolloid; zinc metallopeptidase. 1 Proteolysis Lab , Molecular Biology Institute of Barcelona, CSIC, Barcelona Science Park, Helix Building, c/Baldiri Reixac, 15-21, E-08028 Barcelona , Spain Introduction: a short historical background 2 Institute of Zoology , Cell and Matrix Biology, Johannes Gutenberg University, Johannes-von-M ü ller-Weg 6, The fi rst report on the digestive protease astacin from the D-55128 Mainz , Germany European freshwater crayfi sh, Astacus astacus L. – then termed ‘ crayfi sh small-molecule protease ’ or ‘ Astacus pro- * Corresponding authors tease ’ – dates back to the late 1960s (Sonneborn et al. , 1969 ). e-mail: [email protected]; [email protected] Protein sequencing by Zwilling and co-workers in the 1980s did not reveal homology to any other protein (Titani et al. , Abstract 1987 ). Shortly after, the enzyme was identifi ed as a zinc met- allopeptidase (St ö cker et al., 1988 ), and other family mem- The astacins are a family of multi-domain metallopepti- bers emerged. The fi rst of these was bone morphogenetic β dases with manifold functions in metabolism. They are protein 1 (BMP1), a protease co-purifi ed with TGF -like either secreted or membrane-anchored and are regulated growth factors termed bone morphogenetic proteins due by being synthesized as inactive zymogens and also by co- to their capacity to induce ectopic bone formation in mice localizing protein inhibitors.
    [Show full text]
  • MMP-9) on the Metastatic Behavior of Oropharyngeal Cancer
    ANTICANCER RESEARCH 25: 4129-4134 (2005) Influence of Matrix Metalloproteinase 9 (MMP-9) on the Metastatic Behavior of Oropharyngeal Cancer A.A. DÜNNE1, A. GRÖBE1, A.M. SESTERHENN1, P. BARTH2, C. DALCHOW1 and J.A. WERNER1 1Department of Otolaryngology, Head and Neck Surgery and 2Department of Pathology, Philipps University of Marburg, Marburg, Germany Abstract. Background: The role of the single matrix molecular biological factors have been proposed for their metalloproteinases (MMPs) in the metastatic process of possible impact on the lymphogenic metastatic process. squamous cell carcinomas (SCC) is still obscure. Materials In this context, special interest is being paid to a better and Methods: The MMP-9 expression was described knowledge of the matrix metalloproteinases (MMPs) (5), immunohistochemically in 105 patients (40-79 years of age, that are involved in the physiology of reconstruction and mean: 57.84 years; 84 male, 21 female) suffering from renewal processes of surrounding tissue. In particular, MMP- oropharyngeal cancer (22x T1, 31x T2, 24x T3, 28x T4) with 2 and -9, originating from the subfamily of gelatinases, seem different neck stages (41x N0, 6x N1, 54x N2, 4x N3 neck). to play important roles in the metastatic process of several Results: A significant correlation between MMP-9 expression carcinomas (6, 7), including not only SCC of the head and and T stage (p<0.05), N stage (r=0.55, p<0.01) and UICC neck (8), but also malignant tumors of the lung (9), the stage (r=0.55, p<0.01) was revealed. Most remarkable was the prostate (10), the colon (11) and the bladder (12).
    [Show full text]
  • Identification of the Membrane-Type Matrix Metalloproteinase MT1-MMP
    Journal of Cell Science 110, 589-596 (1997) 589 Printed in Great Britain © The Company of Biologists Limited 1997 JCS9564 Identification of the membrane-type matrix metalloproteinase MT1-MMP in osteoclasts Takuya Sato1, Maria del Carmen Ovejero1, Peng Hou1, Anne-Marie Heegaard1, Masayoshi Kumegawa2, Niels Tækker Foged1 and Jean-Marie Delaissé1,* 1Department of Basic Research, Center for Clinical & Basic Research, Ballerup Byvej 222, DK-2750 Ballerup, Denmark 2The First Department of Oral Anatomy, Meikai University School of Dentistry, Keyakidai 1-1, Sakado, Saitama 350-02, Japan *Author for correspondence SUMMARY The osteoclasts are the cells responsible for bone resorp- on bone sections showed that MT1-MMP is expressed also tion. Matrix metalloproteinases (MMPs) appear crucial in osteoclasts in vivo. Antibodies recognizing MT1-MMP for this process. To identify possible MMP expression in reacted with specific plasma membrane areas corre- osteoclasts, we amplified osteoclast cDNA fragments sponding to lamellipodia and podosomes involved, respec- having homology with MMP genes, and used them as a tively, in migratory and attachment activities of the osteo- probe to screen a rabbit osteoclast cDNA library. We clasts. These observations highlight how cells might bring obtained a cDNA of 1,972 bp encoding a polypeptide of MT1-MMP into contact with focal points of the extracel- 582 amino acids that showed more than 92% identity to lular matrix, and are compatible with a role of MT1- human, mouse, and rat membrane-type 1 MMP (MT1- MMP in migratory and attachment activities of the osteo- MMP), a cell surface proteinase believed to trigger cancer clast. cell invasion. By northern blotting, MT1-MMP was found to be highly expressed in purified osteoclasts when compared with alveolar macrophages and bone stromal Key words: Osteoclast, MMP-14, MT1-MMP, Membrane proteinase, cells, as well as with various tissues.
    [Show full text]
  • Matrix Metalloproteinase -9, Human Recombinant, Expressed in Transfected Cells
    Matrix Metalloproteinase -9, human recombinant, expressed in transfected cells Catalog Number M4809 Storage Temperature –70 °C EC 3.4.24.35 All MMPs are synthesized as proenzymes, and most of Synonyms: MMP-9; Gelatinase-B; 95 kDa Gelatinase them are secreted from the cells as proenzymes. Thus, the activation of these proenzymes is a critical step that Product Description leads to extracellular matrix breakdown. Human Matrix Metalloproteinase-9 (MMP-9) is a matrix metalloproteinase that has been substrate-affinity MMPs are considered to play an important role in purified from transfected cells (pro and active forms). wound healing, apoptosis, bone elongation, embryo development, uterine involution, angiogenesis,5 and Matrix Metalloproteinase-9 (MMP-9) may be used in tissue remodeling, and in diseases such as multiple various immunochemical techniques such as sclerosis,3,6 Alzheimer’s,3 malignant gliomas,3 lupus, immunoblotting, ELISA, enzyme kinetics assays, and arthritis, periodontis, glomerulonephritis, athero- substrate assays. This preparation consists primarilay sclerosis, tissue ulceration, and in cancer cell invasion of the proenzyme form, with a small amount of active and metastasis.7 Numerous studies have shown that enzyme and TIMP present. The following bands may be there is a close association between expression of detected: various members of the MMP family by tumors and MMP-9 Proenzyme (>85%) at ~88 kDa non-reduced their proliferative and invasive behavior and metastatic and 92 kDa reduced potential. minor band (MMP-9 dimer) at ~180 kDa intermediate active form (very small amounts) at The tissue inhibitors of metalloproteinases (TIMPs) are ~84 kDa non-reduced naturally occurring proteins that specifically inhibit mature active form (very small amounts) at 82 kDa matrix metalloproteinases and regulate extracellular non-reduced matrix turnover and tissue remodeling by forming tight - TIMP-1 (~10%) at 28 kDa binding inhibitory complexes with the MMPs.
    [Show full text]
  • Expression of Matrix Metalloproteinase (MMP)-2 and MMP-9 in Breast Cancer with a Special Reference to Activator Protein-2, HER2, and Prognosis
    Vol. 10, 7621–7628, November 15, 2004 Clinical Cancer Research 7621 Expression of Matrix Metalloproteinase (MMP)-2 and MMP-9 in Breast Cancer with a Special Reference to Activator Protein-2, HER2, and Prognosis Johanna M. Pellikainen,1,2,3 Kirsi M. Ropponen,2 positive disease. In the univariate survival analysis, positive Vesa V. Kataja,3 Jari K. Kellokoski,4 stromal MMP-9 predicted shorter recurrence-free survival and breast cancer-related survival (0.0389 ؍ RFS; P) 1,2,3,6 5 ؉ ؍ Matti J. Eskelinen, and Veli-Matti Kosma 1 (BCRS; P 0.0081) in ER disease, especially in the sub- ؍ > Department of Pathology and Forensic Medicine, University of ؉ 2 3 group of ER tumors of 2 cm in diameter (T1; P 0.0031 ؍ ,Kuopio, Kuopio, Finland; Departments of Pathology, Oncology 4Otorhinolaryngology, Oral and Maxillofacial Unit, and 5Surgery, for RFS, and P 0.0089 for BCRS). High MMP-9 expres- in the (0.0351 ؍ Kuopio University Hospital, Kuopio, Finland; and 6Department of sion in cancer cells predicted longer RFS (P Pathology, Centre for Laboratory Medicine, University of Tampere whole patient group. In the multivariate analysis of the and Tampere University Hospital, Tampere, Finland whole patient group, the independent predictors of shorter RFS were reduced MMP-9 expression in carcinoma cells ؍ ؍ ABSTRACT (P 0.0248), HER2 overexpression (P 0.0001), and ad- -Shorter BCRS was pre .(0.0002 ؍ vanced-stage disease (P Purpose: In the present study, we investigated the ex- dicted by advanced-stage disease (P < 0.0001). pression and prognostic value of matrix metalloproteinase Conclusions: Expression of MMP-2 and MMP-9 in (MMP)-2 and MMP-9 in breast cancer as well as their breast cancer seems to be partly related to expression of relation to transcription factor activator protein (AP)-2 AP-2 and HER2.
    [Show full text]
  • Methods for Detection of Matrix Metalloproteinases As Biomarkers in Cardiovascular Disease Viorica Lopez-Avila
    The University of San Francisco USF Scholarship: a digital repository @ Gleeson Library | Geschke Center Biology Faculty Publications Biology 2008 Methods for Detection of Matrix Metalloproteinases as Biomarkers in Cardiovascular Disease Viorica Lopez-Avila Juliet Spencer University of San Francisco, [email protected] Follow this and additional works at: http://repository.usfca.edu/biol_fac Part of the Biology Commons, and the Cardiovascular Diseases Commons Recommended Citation Lopez-Avila, V and Spencer, JV. Methods for Detection of Matrix Metalloproteinases as Biomarkers in Cardiovascular Disease. Clinical Medicine Insights: Cardiology 2008:2 75-87. This Article is brought to you for free and open access by the Biology at USF Scholarship: a digital repository @ Gleeson Library | Geschke Center. It has been accepted for inclusion in Biology Faculty Publications by an authorized administrator of USF Scholarship: a digital repository @ Gleeson Library | Geschke Center. For more information, please contact [email protected]. REVIEW Methods for Detection of Matrix Metalloproteinases as Biomarkers in Cardiovascular Disease Viorica Lopez-Avila1 and Juliet V. Spencer2 1Agilent Technologies, Santa Clara, CA 95051, U.S.A. 2University of San Francisco, San Francisco, CA 94403, U.S.A. Abstract: Matrix metalloproteinases (MMPs) are a family of zinc-dependent proteolytic enzymes that degrade extracel- lular matrix (ECM) components like collagen, fi bronectin, and laminin. While this activity is important for normal develop- ment, morphogenesis, and wound healing, deregulation of MMP activity has been implicated in a number of cardiovascular diseases, including congenital heart defects, atherosclerosis, myocardial infarction, and congestive heart failure. MMPs are good potential diagnostic indicators of cardiovascular disease, but current detection methods are time consuming and quite laborious.
    [Show full text]
  • Matrix Metalloproteinase-3, Catalytic Domain (M9320)
    Matrix Metalloproteinase-3, Catalytic Domain human, recombinant expressed in E. coli Catalog Number M9320 Storage Temperature -70 °C EC 3.4.24.17 Matrix Metalloproteinase-3 (MMP-3) degrades a wide range of substrates, including gelatin, type IV, V, IX, Synonyms: MMP-3; Stromelysin-1; Transin; and X collagens, elastin, laminin, vitronectin, casein, Proteoglycanase fibronectin, proteoglycans, aggrecan, myelin basic protein, and a-1-antitrypsin.8,9 MMP-3 can be induced Product Description by cytokines IL-1b and TNF-a, by growth factors EGF, The matrix metalloproteinases (MMPs) are a family of PDGF, and IGFBP-3, and by the tumor promotor PMA. at least eighteen secreted and membrane-bound zinc- Its expression is inhibited by TGF-b and by all-trans endopeptidases. Collectively, these enzymes can retinoic acid (RA). The MMP-3 substrate repertoire degrade all the components of the extracellular matrix, extends beyond the extracellular matrix proteins and including fibrillar and non-fibrillar collagens, fibro- suggests that MMP-3 has additional roles other than in nectin, laminin and basement membrane glyco- direct tissue remodeling (i.e., enzyme cascades and proteins. In general, a signal peptide, a propeptide, cytokine regulation). and a catalytic domain containing the highly conserved zinc-binding site characterize the structure of the MMP-3 does not cleave the triple helical region of the MMPs. In addition, fibronectin-like repeats, a hinge interstitial collagens, which is a characteristic that region, and a C-terminal hemopexin-like domain allow distinguishes the stromelysins from the collagenases. categorization of MMPs into the collagenase, gela- Structurally, MMP-3 is divided into several distinct tinase, stomelysin and membrane-type MMP sub- domains: a pro-domain which is cleaved upon 1-4 families.
    [Show full text]
  • Gelatinase B/Matrix Metalloproteinase-9 Is a Phase-Specific Effector Molecule, Independent from Fas, in Experimental Autoimmune Encephalomyelitis
    bioRxiv preprint doi: https://doi.org/10.1101/321661; this version posted May 14, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Gelatinase B/matrix metalloproteinase-9 is a phase-specific effector molecule, independent from Fas, in experimental autoimmune encephalomyelitis Estefania Ugarte-Berzal1, Nele Berghmans2, Lise Boon1, Erik Martens1, Jennifer Vandooren1, Bénédicte Cauwe1, Greet Thijs1, Paul Proost2, Jo Van Damme2, Ghislain Opdenakker1 1,2 Rega Institute for Medical Research, 1Laboratory of Immunobiology, 2 Laboratory of Molecular Immunology, Department of Microbiology and Immunology, University of Leuven, KU Leuven, Belgium. Abstract Gelatinase B/matrix metalloproteinase-9 (MMP-9) triggers multiple sclerosis (MS) and the animal model of experimental autoimmune encephalomyelitis (EAE) by the breakdown of the blood-brain barrier. Interestingly, MMP-9 is beneficial in systemic autoimmunity caused by Fas-deficiency. Fas-deficient (faslpr) and Fas-ligand-deficient mice are protected against EAE. We here investigated the interaction between Fas and MMP-9 in the setting of induction of EAE and compared short- and long-term effects. We provoked EAE with myelin oligodendrocyte glycoprotein (MOG) peptide and compared EAE development in four genotypes (wild-type (WT), single knockout mmp-9-/-, faslpr, and mmp-9-/-/faslpr) and monitored leukocytes, cytokines and chemokines as immunological parameters. As expected, faslpr mice were resistant against EAE induction, whereas MMP-9 single knockout mice were not. In the double mmp-9-/-/ faslpr mice the effects on disease scores pointed to independent rather than interrelated disease mechanisms.
    [Show full text]