title: Role for chondroitin sulfate glycosaminoglycan in NEDD9-mediated breast cancer cell growth.
abstract: There are lines of evidence demonstrating that NEDD9 (Cas-L, HEF-1) plays a key role in the development, progression, and metastasis of breast cancer cells. We previously reported that NEDD9 plays a critical role for promoting migration and growth of MDA-MB-231. In order to further characterize the mechanisms of NEDD9-mediated cancer migration and growth, stable cells overexpressing NEDD9 were generated using HCC38 as a parental cell line which expresses low level of endogenous NEDD9. Microarray studies demonstrated that core proteins of CD44 and Serglycin were markedly upregulated in HCC38(NEDD9) cells compared to HCC38(Vector) cells, while those of Syndecan-1, Syndecan-2, and Versican were downregulated in HCC38(NEDD9). Importantly, enzymes generating chondroitin sulfate glycosaminoglycans (CS) such as CHST11, CHST15, and CSGALNACT1 were upregulated in HCC38(NEDD9) compared to HCC38(Vector). Immunofluorescence studies using specific antibody, GD3G7, confirmed the enhanced expression of CS-E subunit in HCC38(NEDD9). Immunoprecipitation and western blotting analysis demonstrated that CS-E was attached to CD44 core protein. We demonstrated that removing CS by chondroitinase ABC significantly inhibited anchorage-independent colony formation of HCC38(NEDD9) in methylcellulose. Importantly, the fact that GD3G7 significantly inhibited colony formation of HCC38(NEDD9) cells suggests that CS-E subunit plays a key role in this process. Furthermore, treatment of HCC38(NEDD9) cells with chondroitinase ABC or GD3G7 significantly inhibited mammosphere formation. Exogenous addition of CS-E enhanced colony formation and mammosphere formation of HCC38 parental and HCC38(Vector) cells. These results suggest that NEDD9 regulates the synthesis and expression of tumor associated glycocalyx structures including CS-E, which plays a key role in promoting and regulating breast cancer progression and metastasis and possibly stem cell phenotypes.
authors.affiliation: Department of Cell Biology, Windber Research Institute, Windber, PA 15963, USA. Electronic address: g.iida@wriwindber.org.
authors.forename: Joji
authors.initials: J
authors.lastname: Iida
authors.affiliation: Department of Cell Biology, Windber Research Institute, Windber, PA 15963, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Dorchak
authors.affiliation: Department of Cell Biology, Windber Research Institute, Windber, PA 15963, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Clancy
authors.affiliation: Department of Cell Biology, Windber Research Institute, Windber, PA 15963, USA.
authors.forename: Juliana
authors.initials: J
authors.lastname: Slavik
authors.affiliation: Clinical Breast Care Project, Henry M, Jackson Foundation for the Advancement of Military Medicine, Windber, PA 15963, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Ellsworth
authors.affiliation: Developmental Neurobiology Section, Cell Biology and Physiology Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Katagiri
authors.affiliation: Department of Biochemistry, School of Medicine, West Virginia University, Morgantown, WV 26506, USA.
authors.forename: Elena N
authors.initials: EN
authors.lastname: Pugacheva
authors.affiliation: Department of Biochemistry, Nijmegen Centre for Molecular Life Science, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.affiliation: Department of Cell Biology, Windber Research Institute, Windber, PA 15963, USA.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Mural
authors.affiliation: Department of Pathology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Rd, Bethesda, MD 20814, USA.
authors.forename: Mary Lou
authors.initials: ML
authors.lastname: Cutler
authors.affiliation: Department of Surgery, Walter-Reed National Military Medical Center, Bethesda, MD 20814, USA.
authors.forename: Craig D
authors.initials: CD
authors.lastname: Shriver
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000096:biosynthesis / Q000378:metabolism; D001943:Breast Neoplasms / Q000473:pathology*; D002465:Cell Movement; D049109:Cell Proliferation; D019765:Chondroitin ABC Lyase / Q000378:metabolism / Q000494:pharmacology; D002809:Chondroitin Sulfates / Q000096:biosynthesis*; D015536:Down-Regulation; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D018960:Hyaluronan Receptors / Q000096:biosynthesis; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D017350:N-Acetylgalactosaminyltransferases / Q000096:biosynthesis; D009362:Neoplasm Metastasis / Q000473:pathology; D010750:Phosphoproteins / Q000096:biosynthesis / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis / Q000378:metabolism; D018874:Spheroids, Cellular / Q000473:pathology*; D015238:Sulfotransferases / Q000096:biosynthesis; D053668:Syndecan-1 / Q000096:biosynthesis; D053669:Syndecan-2 / Q000096:biosynthesis; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D053675:Versicans / Q000096:biosynthesis; D033921:Vesicular Transport Proteins / Q000096:biosynthesis; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000911:Antibodies, Monoclonal; D000941:Antigens; D019765:Chondroitin ABC Lyase; D002809:Chondroitin Sulfates; D018960:Hyaluronan Receptors; D008562:Membrane Glycoproteins; D017350:N-Acetylgalactosaminyltransferases; D010750:Phosphoproteins; D011509:Proteoglycans; D015238:Sulfotransferases; D053668:Syndecan-1; D053669:Syndecan-2; D053675:Versicans; D033921:Vesicular Transport Proteins; D000099216:Chondroitin Sulfate Proteoglycan 4; C497483:CD44 protein, human; C411874:CHST15 protein, human; C099942:NEDD9 protein, human; C508322:VCAN protein, human; C038142:chondroitin sulfate glycosaminoglycan; C075430:serglycin; C472559:chondroitin sulfate N-acetylgalactosaminyltransferase-1; C411659:CHST11 protein, human
keywords: Breast cancer; CD44; Chondroitin sulfate; Growth; Migration; NEDD9