title: An uncoordinated-5 homolog B receptor monoclonal antibody regulates A375 melanoma cell migration.
abstract: Uncoordinated-5 homolog B receptor (UNC5B) was first found to mediate neural chemorepulsive effects by binding to its ligand netrin-1 in the nervous system. Newer evidence indicated that UNC5B also has functions outside the nervous system. In this study, we report on the generation of a monoclonal antibody specific to the outer-membrane immunoglobulin-like domains of UNC5B using the hybridoma technique. Western blot, immunofluorescence, and flow cytometry analyses showed that the antibody specifically bound to UNC5B protein. Interestingly, the antibody blocked the Netrin-1-induced inhibitory effect on the mobility of melanoma A375 cells by wound healing assay and transwell migration assay, whereas it had no effects on cell proliferation measured by CCK-8 assay. Thus, the functional antibody may provide a useful tool for the study of UNC5B expression profiles and functions outside the nervous system.
authors.affiliation: 1 Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University , Suzhou, China .
authors.forename: Yuanling
authors.initials: Y
authors.lastname: Zuo
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Yang
authors.forename: Jun
authors.initials: J
authors.lastname: He
authors.forename: Yiming
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yang
authors.initials: Y
authors.lastname: He
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000276:immunology*; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D000075386:Netrin Receptors; D011956:Receptors, Cell Surface / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D000075386:Netrin Receptors; D011956:Receptors, Cell Surface; C532445:UNC5B protein, human