DOCSLIB.ORG

Endoglin Expression in Breast Tumor Cells Suppresses Invasion and Metastasis and Correlates with Improved Clinical Outcome

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Endoglin Expression in Breast Tumor Cells Suppresses Invasion and Metastasis and Correlates with Improved Clinical Outcome Oncogene (2011) 30, 1046–1058 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc ORIGINAL ARTICLE Endoglin expression in breast tumor cells suppresses invasion and metastasis and correlates with improved clinical outcome LA Henry1, DA Johnson1, D Sarrio´1, S Lee1, PR Quinlan2, T Crook1, AM Thompson2, JS Reis-Filho1 and CM Isacke1 1Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, London, UK and 2Department of Surgery and Molecular Oncology, Ninewells Hospital and Medical School, Dundee, UK Tumor growth factor-b (TGF-b) signaling in cancer has been and metastatic events in late stage carcinomas (Roberts implicated in growth suppression of early lesions and and Wakefield, 2003; Massague, 2008). TGF-b ligands act enhancing tumor cell invasion and metastasis. However, by binding to their cognate transmembrane serine/ the cellular mechanisms that determine this signaling output threonine kinase receptors (Shi and Massague, 2003; Feng in individual tumors are still largely unknown. In endothelial and Derynck, 2005). This results in the activation of the cells, TGF-b signaling is modulated by the TGF-b co- canonical TGF-b pathway mediated by the phosphoryla- receptor endoglin (CD105). Here we demonstrate that tion of Smad proteins and their subsequent translocation endoglin is expressed in a subset of invasive breast cancers to the nucleus where they promote the transcriptional and cell lines and is subject to epigenetic silencing by gene regulation of TGF-b response genes. However, TGF-b can methylation. Endoglin downregulation in non-tumorigenic also signal through a range of other pathways (Moustakas MCF10A breast cells leads to the formation of abnormal and Heldin, 2005; Zhang, 2009) and these alternate acini in 3D culture, but does not promote cell migration or pathways are particularly important in the pro-invasive transformation. In contrast, in the presence of activated responses to TGF-b in cancer (Bhowmick et al., 2001; ErbB2, endoglin downregulation in MCF10A cells leads to Ozdamar et al., 2005). enhanced invasion into a 3D matrix. Consistent with these Endoglin (CD105) is a type I integral transmembrane data, ectopic expression of endoglin in MDA-MB-231 cells glycoprotein that has predominantly been investigated blocks TGF-b-enhanced cell motility and invasion and in the vasculature where it is upregulated during reduces lung colonization in an in vivo metastasis model. angiogenesis (Bernabeu et al., 2007, 2009). In endothe- Unlike endothelial cells, endoglin does not modulate Smad- lial cells, endoglin associates at the cell surface with the mediated TGF-b signaling in breast cells but attenuates the type I and type II TGF-b receptors and has a key role in cytoskeletal remodeling to impair cell migration and modulating downstream signaling to the Smad transdu- invasion. Importantly, in a large cohort of invasive breast cer proteins (Guerrero-Esteo et al., 2002; Lebrin et al., cancers, lack of endoglin expression in the tumor cell 2004). The importance of endoglin acting as a TGF-b compartment correlates with ENG gene methylation and co-receptor in these events is evidenced by the observa- poor clinical outcome. tion that mutations in either ENG or the type I receptor Oncogene (2011) 30, 1046–1058; doi:10.1038/onc.2010.488; ALK1 give rise to the vascular disorder hereditary published online 1 November 2010 hemorrhagic telangiectasia (McAllister et al., 1994; Johnson et al., 1996). From an initial demethylation Keywords: endoglin; CD105; breast cancer; TGF-b; screen, we identified ENG as a candidate gene for MCF10A; ErbB2 epigenetic regulation in breast cancer cell lines. Together with previous reports that endoglin can inhibit the migration of other epithelial tumors (Craft et al., 2007; Perez-Gomez et al., 2007), we hypothesized that Introduction expression of endoglin might function to suppress breast cancer progression. To address this we investigated the The tumor growth factor-b (TGF-b) signaling pathway functional role of endoglin in both in vitro and in vivo has an important role in all stages of breast cancer models, its impact on the TGF-b signaling pathway and progression. It is well established that TGF-b has a dual the prognostic impact and regulation of endoglin function in acting both as a tumor suppressor in the early expression in a large cohort of invasive breast cancers. stage of malignant progression and to promote invasive Results Correspondence: Professor CM Isacke, Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, 237 Fulham Road, Downregulation of endoglin expression enhances breast London SW3 6JB, UK. E-mail: [email protected] cancer cell invasion Received 30 March 2010; revised 31 August 2010; accepted 3 September A panel of cell lines was subject to immunoblotting and 2010; published online 1 November 2010 quantitative real-time PCR (qPCR). Endoglin mRNA Endoglin suppresses breast cancer invasion LA Henry et al 1047 100 Endoglin MCF10A 75 (Long exposure) Endoglin 100 75 (Short exposure) shEng1 shEng5 shEng2 shCont Parental 50 Tubulin 100 Endoglin 75 50 37 ERK1/2 Relative mRNA expression p=0.0145 p=0.0141 p=0.1927 Parental shCont shEng2 % of abnormal acini shEng1 shEng5 shCont shEng1 shEng2 shEng5 Parental Figure 1 Endoglin downregulation enhances abnormal MCF10A acini formation in 3D culture. (a) Endoglin expression analyzed in a series of breast cell lines by immunoblotting (upper panels) or qPCR (lower panel). Data shown are the mean±s.e.m. of RQ relative expression of two independent experiments. (b) Immunoblotting of MCF10A cells stably infected with different shRNA lentiviral constructs against endoglin (shEng) or control (shCont). Note, the shEng2 cell line showed negligible endoglin knockdown and therefore was used as an independent shRNA control. (c) MCF10A cell lines stably expressing control shRNA (shCont, shEng2) or endoglin knockdown (shEng1, shEng5) shRNA constructs were grown in 3D culture and fixed on day 20. Cells were permeabilized and stained for collagen IV and Alexa488 anti-rabbit Ig (green), the Golgi protein b-COP and Alexa555 anti-rat Ig (red) and DAPI (blue). Left panel, representative confocal images. Scale bar, 25 mm. Right panel, mean % of abnormal acini morphology for three independent experiments±s.e.m. and protein expression was readily detected in the non- endoglin lentiviral small hairpin RNAs (shRNAs) tumorigenic breast cell line MCF10A, in a subset of the (shEng1–5) targeting different regions of the ENG gene cancer cell lines (MDA-MB-468, SKBR3, T47D) and in or with control shRNA (shCont) (Figure 1b). Substan- the recently re-classified melanoma line MDA-MB-435 tial knockdown was observed with shEng1 and shEng5 (Figure 1a). This expression was 2.5–5-fold lower than but not with shEng2. The shEng2 and shCont cell lines found in the cultured endothelial cells (Supplementary expressed similar levels of endoglin as the parental line Figure 1). No endoglin expression was detected in and hence were used as two independent controls. In MCF7, Cal51, MDA-MB-231, MDA-MB-453, HCC1954 this 3D culture model, shCont and shEng2 cells were or BT474 cells. Reverse transcription–PCR analysis indistinguishable from the parental MCF10A cells and revealed that in the endoglin-positive cell lines the formed single acini that contained a hollow lumen predominant expression was of the long-endoglin isoform surrounded by a polarized layer of epithelial cells with with little or no expression of the short-endoglin isoform their Golgi orientated inwards (Figure 1c). In contrast, detected (data not shown). the endoglin-knockdown lines (shEng1 and shEng5) To explore a potential role for endoglin as a breast produced abnormally shaped acini in which the outer cancer suppressor, we initiated this study with the non- layer of cells was not fully polarized and the lumena tumorigenic MCF10A cell line. MCF10A cells when were incompletely cleared. Nonetheless, these shEng1 grown on a thick layer of Matrigel form epithelial acini and shEng5 acini, still growth arrested, did not invade (3D culture model), and this system has been used into the Matrigel and were no larger than control extensively to identify pathways that can perturb the structures indicating that the loss of endoglin expression normal breast architecture (Debnath and Brugge, 2005). alone does not result in the transformation of the non- MCF10A cells were stably infected with five individual tumorigenic MCF10A cells. Oncogene Endoglin suppresses breast cancer invasion LA Henry et al 1048 Cell line Expression MCF10A.ErbB2 parental 100% MCF10A.ErbB2 shCont 91.1% MCF10A.ErbB2 shEng1 33.2% MCF10A.ErbB2 shEng2 71.0% MCF10A.ErbB2 shEng5 4.7% p=0.0126 p=0.0428 Parental shEng1 p=0.0403 p=0.0087 p=0.7998 shCont shEng5 % of acini with invasive protrusions -+++ - +++ -+++ +++ -+++- AP1510 --+--+- --+----+- -+- - TGF-β1 shEng2 ---+--+ ---+---+---- + SB431542 Parental shContshEng2 shEng1 shEng5 Figure 2 Endoglin downregulation cooperates with ErbB2 activation to promote invasion in 3D culture. (a) MCF10A.ErbB2 cells were infected with shCont or shEng lentiviral constructs. Stable cell lines were subject to FACS analysis with an isotype control or endoglin antibody. The mean level of fluorescence signal (FITC-A) relative to the parental cells is shown. Note, the shEng2 cell line has negligible endoglin knockdown and therefore was used as an independent shRNA control. (b) MCF10A.ErbB2 cell lines were grown in 3D culture for 5 days±TGF-b1±SB431542±AP1510 dimerizer added from day 4. Acini were stained for laminin V and Alexa488 anti-mouse Ig (green), Alexa546-phalloidin (red), and DAPI (blue). Left panel, representative confocal images of cultures with AP1510 dimerizer added from day 4 in the absence of TGF-b1 and SB431542. Arrowheads indicate invasion protrusions and laminin V degradation. Scale bar, 25 mm. Right panel, mean % of acini with invasive protrusions for three independent experiments±s.e.m. We next tested whether endoglin might function to AP1510-treated cultures produced acini with signifi- modulate the effects of activated oncogenes.
Load more

Recommended publications
  • The Prevalence of MADH4 and BMPR1A Mutations in Juvenile Polyposis and Absence of BMPR2, BMPR1B, and ACVR1 Mutations
    484 ORIGINAL ARTICLE J Med Genet: first published as 10.1136/jmg.2004.018598 on 2 July 2004. Downloaded from The prevalence of MADH4 and BMPR1A mutations in juvenile polyposis and absence of BMPR2, BMPR1B, and ACVR1 mutations J R Howe, M G Sayed, A F Ahmed, J Ringold, J Larsen-Haidle, A Merg, F A Mitros, C A Vaccaro, G M Petersen, F M Giardiello, S T Tinley, L A Aaltonen, H T Lynch ............................................................................................................................... J Med Genet 2004;41:484–491. doi: 10.1136/jmg.2004.018598 Background: Juvenile polyposis (JP) is an autosomal dominant syndrome predisposing to colorectal and gastric cancer. We have identified mutations in two genes causing JP, MADH4 and bone morphogenetic protein receptor 1A (BMPR1A): both are involved in bone morphogenetic protein (BMP) mediated signalling and are members of the TGF-b superfamily. This study determined the prevalence of mutations See end of article for in MADH4 and BMPR1A, as well as three other BMP/activin pathway candidate genes in a large number authors’ affiliations ....................... of JP patients. Methods: DNA was extracted from the blood of JP patients and used for PCR amplification of each exon of Correspondence to: these five genes, using primers flanking each intron–exon boundary. Mutations were determined by Dr J R Howe, Department of Surgery, 4644 JCP, comparison to wild type sequences using sequence analysis software. A total of 77 JP cases were University of Iowa College sequenced for mutations in the MADH4, BMPR1A, BMPR1B, BMPR2, and/or ACVR1 (activin A receptor) of Medicine, 200 Hawkins genes. The latter three genes were analysed when MADH4 and BMPR1A sequencing found no mutations.
    [Show full text]
  • Human Endoglin/CD105 Quantikine
    Quantikine® ELISA Human Endoglin/CD105 Immunoassay Catalog Number DNDG00 For the quantitative determination of human Endoglin concentrations in cell culture supernates, serum, and plasma. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures. TABLE OF CONTENTS SECTION PAGE INTRODUCTION .....................................................................................................................................................................1 PRINCIPLE OF THE ASSAY ...................................................................................................................................................2 LIMITATIONS OF THE PROCEDURE .................................................................................................................................2 TECHNICAL HINTS .................................................................................................................................................................2 MATERIALS PROVIDED & STORAGE CONDITIONS ...................................................................................................3 OTHER SUPPLIES REQUIRED .............................................................................................................................................3 PRECAUTIONS .........................................................................................................................................................................4 SAMPLE COLLECTION & STORAGE
    [Show full text]
  • The Role of the TGF- Co-Receptor Endoglin in Cancer
    1 The role of the TGF- co-receptor endoglin in cancer Eduardo Pérez-Gómez1,†, Gaelle del Castillo1, Juan Francisco Santibáñez2, Jose Miguel López-Novoa3, Carmelo Bernabéu4 and 1,* Miguel Quintanilla . 1Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas (CSIC)-Universidad Autónoma de Madrid, 28029-Madrid, Spain; 2Institute for Medical Research, University of Belgrado, Belgrado, Serbia; 3Instituto Reina Sofía de Investigación Nefrológica, Departamento de Fisiología y Farmacología, Universidad de Salamanca, Salamanca, Spain; 4Centro de Investigaciones Biológicas, CSIC, and CIBER de Enfermedades Raras (CIBERER), Madrid, Spain. E-mails: [email protected]; [email protected]; [email protected]; [email protected]; [email protected]; [email protected] † Current address: Departamento de Bioquímica y Biología Molecular I, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain *Corresponding author 2 ABSTRACT Endoglin (CD105) is an auxiliary membrane receptor of transforming growth factor- (TGF-) that interacts with type I and type II TGF- receptors and modulates TGF- signalling. Mutations in endoglin are involved in Hereditary Hemorrhagic Telangiectasia type I, a disorder characterized by cutaneous telangiectasias, epistaxis (nosebleeds) and major arteriovenous shunts, mainly in liver and lung. Endoglin is overexpressed in the tumor-associated vascular endothelium where it modulates angiogenesis. This feature makes endoglin a promising target for antiangiogenic cancer therapy. Recent studies on human and experimental models of carcinogenesis point to an important tumor cell-autonomous role of endoglin by regulating proliferation, migration, invasion and metastasis. These studies suggest that endoglin behaves as a suppressor of malignancy in experimental and human carcinogenesis. In this review, we evaluate the implication of endoglin in tumor development underlying studies developed in our laboratories in recent years.
    [Show full text]
  • Molecular Classification of Patients with Unexplained Hamartomatous and Hyperplastic Polyposis
    ORIGINAL CONTRIBUTION Molecular Classification of Patients With Unexplained Hamartomatous and Hyperplastic Polyposis Kevin Sweet, MS, CGC Context Significant proportions of patients with hamartomatous polyposis or with Joseph Willis, MD hyperplastic/mixed polyposis remain without specific clinical and molecular diagnosis Xiao-Ping Zhou, MD, PhD or present atypically. Assigning a syndromic diagnosis is important because it guides management, especially surveillance and prophylactic surgery. Carol Gallione, PhD Objective To systematically classify patients with unexplained hamartomatous or hy- Takeshi Sawada, MD, PhD perplastic/mixed polyposis by extensive molecular analysis in the context of central Pia Alhopuro, MD rereview of histopathology results. Sok Kean Khoo, PhD Design, Setting, and Patients Prospective, referral-based study of 49 unrelated patients from outside institutions (n=28) and at a comprehensive cancer center (n=21), Attila Patocs, MD, PhD conducted from May 2, 2002, until December 15, 2004. Germline analysis of PTEN, Cossette Martin, PhD BMPR1A, STK11 (sequence, deletion), SMAD4, and ENG (sequence), specific exon screen- Scott Bridgeman, BSc ing of BRAF, MYH, and BHD, and rereview of polyp histology results were performed. John Heinz, PhD Main Outcome Measures Molecular, clinical, and histopathological findings in pa- tients with unexplained polyposis. Robert Pilarski, MS, CGC Results Of the 49 patients, 11 (22%) had germline mutations. Of 14 patients with Rainer Lehtonen, BSc juvenile polyposis, 2 with early-onset disease had mutations in ENG, encoding endo- Thomas W. Prior, PhD glin, previously only associated with hereditary hemorrhagic telangiectasia; 1 had hemi- zygous deletion encompassing PTEN and BMPR1A; and 1 had an SMAD4 mutation. Thierry Frebourg, MD, PhD One individual previously classified with Peutz-Jeghers syndrome had a PTEN dele- Bin Tean Teh, MD, PhD tion.
    [Show full text]
  • Differential Expression of Endoglin in Human Melanoma Cells Expressing the V3 Isoform of Versican by Microarray Analysis
    MOLECULAR MEDICINE REPORTS 3: 1035-1039, 2010 Differential expression of endoglin in human melanoma cells expressing the V3 isoform of versican by microarray analysis LAIA MIQUEL-SERRA, DANIEL HERNÁNDEZ, MARÍA-JOSÉ DOCAMPO and ANNA BASSOLS Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Spain Received May 28, 2010; Accepted August 13, 2010 DOI: 10.3892/mmr.2010.357 Abstract. Versican is a large chondroitin sulfate proteoglycan overexpression reduced tumor growth rate in mice, but favored produced by several tumor types, including malignant mela- the appearance of secondary tumors (12). noma, which exists as four different splice variants. The large Endoglin (ENG, CD105) is a cell surface component of the isoforms V0 and V1 promote melanoma cell proliferation. transforming growth factor (TGF)-β receptor complex (13), We previously described that overexpression of the short V3 which is highly expressed in the tumor-associated vascular isoform in MeWo human melanoma cells markedly reduced endothelium (14) and in tumor cells. The expression of tumor cell growth in vitro and in vivo, but favored the appear- endoglin appears to play an important role in cancer progres- ance of secondary tumors. This study aimed to elucidate the sion, influencing cell proliferation, motility, invasiveness and mechanisms of V3 by identifying differentially expressed tumorigenicity (15). genes between parental and V3-expressing MeWo melanoma In the present study, we used a high-throughput approach cells using microarray analysis. V3 expression significantly to gain further insight into the mechanism by which the V3 reduced the expression of endoglin, a transforming growth isoform exerts its effects on tumor progression.
    [Show full text]
  • Modulating the Crosstalk Between the Tumor and the Microenvironment Using Sirna: a Flexible Strategy for Breast Cancer Treatment
    cancers Review Modulating the Crosstalk between the Tumor and the Microenvironment Using SiRNA: A Flexible Strategy for Breast Cancer Treatment 1,2, 2, 1,2, Giuseppina Roscigno y, Iolanda Scognamiglio y, Francesco Ingenito y, 2, 1,2 1 2,3, Rosario Vincenzo Chianese y, Francesco Palma , Alan Chan and Gerolama Condorelli * 1 Percuros BV, Albinusdreef 2, 2333 ZA Leiden, The Netherlands; [email protected] (G.R.); [email protected] (F.I.); [email protected] (F.P.); [email protected] (A.C.) 2 Department of Molecular Medicine and Medical Biotechnology, “Federico II” University of Naples, Via Pansini 5, 80131 Naples, Italy; [email protected] (I.S.); [email protected] (R.V.C.) 3 Istituto per l’Endocrinologia e l’Oncologia Sperimentale “G. Salvatore” (IEOS), Consiglio Nazionale delle Ricerche (CNR), 80131 Naples, Italy * Correspondence: [email protected]; Tel.: +39-08-1545-2921 These authors contributed equally. y Received: 30 October 2020; Accepted: 4 December 2020; Published: 13 December 2020 Simple Summary: With this review we aimed to collect the most relevant scientific findings regarding siRNA therapeutic tools against breast cancer microenvironment. Remarkably, breast cancer treatments have been redirected towards the tumor microenvironment components, mainly involved in patients’ relapse and pharmacological resistance. Therefore, siRNAs represent a promising strategy to jeopardize the tumor microenvironment interplay thanks to their non-toxic and specific effects. Abstract: Tumorigenesis is a complex and multistep process in which sequential mutations in oncogenes and tumor-suppressor genes result in enhanced proliferation and apoptosis escape. Over the past decades, several studies have provided evidence that tumors are more than merely a mass of malignant cancer cells, with the tumor microenvironment (TME) also contributing to cancer progression.
    [Show full text]
  • The Targeting of Endoglin on Vascular Endothelial Cells Affects The
    921 Original Article The targeting of endoglin on vascular endothelial cells affects the infiltration of M2 macrophages into the breast cancer microenvironment by modulating the interleukin-6 (IL-6) level Long Zhang1, Bin Yang1, Xiaoli Li1, Yunjing Zhang1, Jinping Zhao1, Wenbin Wang1, Xiaohui Yu2, Zhenhua Zhai1, Hongzhi Sun3 1The Laboratory of Tumour Angiogenesis and Microenvironment, 2Department of Oncology, 3Department of General Surgery, Cancer Centre, The First Hospital Affiliated to Jinzhou Medical University, Jinzhou 121000, China Contributions: (I) Conception and design: L Zhang, Z Zhai, B Yang; (II) Administrative support: H Sun, Z Zhai; (III) Provision of study materials or patients: L Zhang, B Yang, J Zhao; (IV) Collection and assembly of data: All authors; (V) Data analysis and interpretation: L Zhang, B Yang, Z Zhai; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. Correspondence to: Hongzhi Sun. Department of General Surgery, Cancer Centre, The First Hospital Affiliated to Jinzhou Medical University, Jinzhou 121000, China. Email: [email protected]; Zhenhua Zhai. The Laboratory of Tumour Angiogenesis and Microenvironment, Cancer Centre, The First Hospital Affiliated to Jinzhou Medical University, 5-2 Renmin Street, Guta District, Jinzhou 121000, China. Email: [email protected]. Background: The effect and underlying mechanisms of endoglin on angiogenesis and immunity during tumour progression were investigated. Methods: Differences in the growth of established EO771 breast tumours between
    [Show full text]
  • BD Stem Cell Resource Markers of Self-Renewal and Differentiation
    BD Stem Cell Resource Markers of Self-Renewal and Differentiation 23-9953-02 Self Renewing Pathway TGF-β/Activin Sperm Liver Thymus Pancreas Thyroid Lung Intestine Wnt BMP-SMad Erk-MAPK Skin and Hair JAK-STAT Wnt TGF-β/Activin BMP4 bFGF LIF LIFR gp130 Egg p p p Jak β-catenin Smads 175 MEK/ERK Stat3 C-myc Sox2 Oct4 Nanog Smads 2/3 Neural Crest Primordial Germ Cell Stem Cell Neural Stem Cell Self Renewal Ectoderm Embryonic Stem Cell or Induced Pluripotent Endoderm Neurons, Glia Stem Cell Smooth Muscle Cardiac Tissue Chondrocytes Glial Restricted Progenitor Osteocytes Neuronal Type 1 Astrocyte Oligodendrocyte Restricted Progenitor Progenitor Mesoderm Mesenchymal Stem Cell Type 2 Astrocyte Endothelium Oligodendrocyte Neuron Heart Skeletal Muscle Kidney Smooth Muscle Myoblast Adipocyte (Fat) Chondrocyte Fibroblast Osteoblast (Bone) Hemangioblast (Cartilage) Myotube (Muscle) Plasmacytoid Dendritic Cell Hematopoietic Stem Cell Committed Lymphoid Progenitor Monoblast Megakaryoblast Proerythroblast Pre-NK Cell Thymocyte Pre B Cell Myeloblast Erythroblast Progranulocyte Monocyte Myeloid Dendrtitic Cell Megakaryocyte NK Lymphoblast T-Lymphoblast B-Lymphoblast Normoblast Neutrophilic Eosinophilic Basophilic Myelocyte Myelocyte Myelocyte NK Cell T Cell B Cell Macrophage Thrombocytes Reticulocyte Neutrophilic Eosinophilic Basophilic (Platelets) Band Cell Band Cell Band Cell Plasma Cell Erythrocyte Neutrophil Eosinophil Basophil (Red Blood Cell) White colored markers are available from BD Biosciences Ectoderm Markers Embryonic Stem Cells Mesoderm Markers
    [Show full text]
  • Endoglin in the Spotlight to Treat Cancer
    International Journal of Molecular Sciences Review Endoglin in the Spotlight to Treat Cancer Teresa González Muñoz 1,† , Ana Teresa Amaral 2,*,†, Pilar Puerto-Camacho 2,†,Héctor Peinado 1,*,‡ and Enrique de Álava 2,3,*,‡ 1 Microenvironment and Metastasis Group, Department of Molecular Oncology, Spanish National Cancer Research Center (CNIO), 28029 Madrid, Spain; [email protected] 2 Molecular Pathology of Sarcomas, Institute of Biomedicine of Sevilla (IBiS), Virgen del Rocio University Hospital, CSIC, University of Sevilla, CIBERONC, 41013 Seville, Spain; [email protected] 3 Department of Normal and Pathological Cytology and Histology, School of Medicine, University of Seville, 41009 Seville, Spain * Correspondence: [email protected] (A.T.A.); [email protected] (H.P.); [email protected] (E.d.Á.) † These authors contributed equally to this work. ‡ These authors shared senior authorship. Abstract: A spotlight has been shone on endoglin in recent years due to that fact of its potential to serve as both a reliable disease biomarker and a therapeutic target. Indeed, endoglin has now been assigned many roles in both physiological and pathological processes. From a molecular point of view, endoglin mainly acts as a co-receptor in the canonical TGFβ pathway, but also it may be shed and released from the membrane, giving rise to the soluble form, which also plays important roles in cell signaling. In cancer, in particular, endoglin may contribute to either an oncogenic or a non-oncogenic phenotype depending on the cell context. The fact that endoglin is expressed by neoplastic and non-neoplastic cells within the tumor microenvironment suggests new possibilities Citation: González Muñoz, T.; for targeted therapies.
    [Show full text]
  • Mouse Endoglin/CD105 Antibody Antigen Affinity-Purified Polyclonal Goat Igg Catalog Number: AF1320
    Mouse Endoglin/CD105 Antibody Antigen Affinity-purified Polyclonal Goat IgG Catalog Number: AF1320 DESCRIPTION Species Reactivity Mouse Specificity Detects mouse Endoglin in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross­reactivity with recombinant human Endoglin is observed. Source Polyclonal Goat IgG Purification Antigen Affinity­purified Immunogen Mouse myeloma cell line NS0­derived recombinant mouse Endoglin/CD105 Glu27­Gly581 Accession # Q8K100 Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Western Blot 0.1 µg/mL Recombinant Mouse Endoglin/CD105 Fc Chimera (Catalog # 1320­EN) Flow Cytometry 2.5 µg/106 cells MS­1 mouse cell line Immunocytochemistry 5­15 µg/mL See Below Immunohistochemistry 5­15 µg/mL See Below DATA Immunocytochemistry Immunocytochemistry Endoglin/CD105 in MS­1 Endoglin/CD105 in Rat Mesenchymal Mouse Cell Line. Stem Cells. Endoglin/CD105 was detected in Endoglin/CD105 was detected in immersion fixed rat mesenchymal stem cells immersion fixed MS­1 mouse using Goat Anti­Mouse Endoglin/CD105 pancreatic islet endothelial cell Antigen Affinity­purified Polyclonal Antibody line using Goat Anti­Mouse (Catalog # AF1320) at 10 µg/mL for 3 hours Endoglin/CD105 Antigen Affinity­ at room temperature. Cells were stained purified Polyclonal Antibody using the NorthernLights™ 557­conjugated (Catalog # AF1320) at 10 µg/mL Anti­Goat IgG Secondary Antibody (red; for 3 hours at room temperature.
    [Show full text]
  • Endoglin As an Adhesion Molecule in Mature and Progenitor Endothelial Cells: a Function Beyond TGF-Β
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Digital.CSIC MINI REVIEW published: 30 January 2019 doi: 10.3389/fmed.2019.00010 Endoglin as an Adhesion Molecule in Mature and Progenitor Endothelial Cells: A Function Beyond TGF-β Elisa Rossi 1,2*, Carmelo Bernabeu 3 and David M. Smadja 1,2,4,5 1 Université Paris Descartes, Sorbonne Paris Cité, Paris, France, 2 Inserm UMR-S1140, Paris, France, 3 Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid, Spain, 4 Department of Hematology, AP-HP, Hôpital Européen Georges Pompidou, Paris, France, 5 Laboratory of Biosurgical Research, Carpentier Foundation, Hôpital Européen Georges Pompidou, Paris, France Endoglin (ENG) is a transmembrane glycoprotein expressed on endothelial cells that functions as a co-receptor for several ligands of the transforming growth factor beta (TGF-β) family. ENG is also a recognized marker of angiogenesis and mutations in the endoglin gene are responsible for Hereditary Hemorrhagic Telangiectasia (HHT) type 1, a vascular disease characterized by defective angiogenesis, arteriovenous malformations, Edited by: telangiectasia, and epistaxis. In addition to its involvement in the TGF-β family signaling Peter S. Steyger, pathways, several lines of evidence suggest that the extracellular domain of ENG has Oregon Health and Science University, a role in integrin-mediated cell adhesion via its RGD motif. Indeed, we have described United States a role for endothelial ENG in leukocyte trafficking and extravasation via its binding to Reviewed by: Eleni Papakonstantinou, leukocyte integrins. We have also found that ENG is involved in vasculogenic properties of Aristotle University of Thessaloniki, endothelial progenitor cells known as endothelial colony forming cells (ECFCs).
    [Show full text]
  • Recombinant Human Endoglin/CD105 Fc Chimera
    Recombinant Human Endoglin/CD105 Fc Chimera Catalog Number: 6578-EN DESCRIPTION Source Chinese Hamster Ovary cell line, CHO­derived Human Endoglin Human IgG (Met1­Gly586) IEGRMD 1 (Pro100­Lys330) Accession # P17813 N­terminus C­terminus N­terminal Sequence Glu26 Analysis Predicted Molecular 87.3 kDa (monomer) Mass SPECIFICATIONS SDS­PAGE 100­115 kDa, reducing conditions Activity Measured by its ability to inhibit BMP­10­induced alkaline phosphatase production by MC3T3­E1 mouse preosteoblast cells. The ED50 for this effect is typically 0.3­1.2 μg/mL. Endotoxin Level <0.10 EU per 1 μg of the protein by the LAL method. Purity >90%, by SDS­PAGE under reducing conditions and visualized by silver stain. Formulation Lyophilized from a 0.2 μm filtered solution in PBS. See Certificate of Analysis for details. PREPARATION AND STORAGE Reconstitution Reconstitute at 250 μg/mL in PBS. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. Stability & Storage Use a manual defrost freezer and avoid repeated freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 3 months, ­20 to ­70 °C under sterile conditions after reconstitution. BACKGROUND Endoglin (CD105) is a 90 kDa type I transmembrane glycoprotein of the zona pellucida (ZP) family of proteins (1­3). Endoglin and betaglycan/TβRIII are type III receptors for TGF beta superfamily ligands, sharing 71% aa identity with within the transmembrane (TM) and cytoplasmic domains. Endoglin is highly expressed on proliferating vascular endothelial cells, chondrocytes, and syncytiotrophoblasts of term placenta, with lower amounts on hematopoietic, mesenchymal and neural crest stem cells, activated monocytes, and lymphoid and myeloid leukemic cells (2­5).
    [Show full text]