title: CCL5/CCR5 axis induces vascular endothelial growth factor-mediated tumor angiogenesis in human osteosarcoma microenvironment.
abstract: Chemokines modulate angiogenesis and metastasis that dictate cancer development in tumor microenvironment. Osteosarcoma is the most frequent bone tumor and is characterized by a high metastatic potential. Chemokine CCL5 (previously called RANTES) has been reported to facilitate tumor progression and metastasis. However, the crosstalk between chemokine CCL5 and vascular endothelial growth factor (VEGF) as well as tumor angiogenesis in human osteosarcoma microenvironment has not been well explored. In this study, we found that CCL5 increased VEGF expression and production in human osteosarcoma cells. The conditioned medium (CM) from CCL5-treated osteosarcoma cells significantly induced tube formation and migration of human endothelial progenitor cells. Pretreatment of cells with CCR5 antibody or transfection with CCR5 specific siRNA blocked CCL5-induced VEGF expression and angiogenesis. CCL5/CCR5 axis demonstrably activated protein kinase Cδ (PKCδ), c-Src and hypoxia-inducible factor-1 alpha (HIF-1α) signaling cascades to induce VEGF-dependent angiogenesis. Furthermore, knockdown of CCL5 suppressed VEGF expression and attenuated osteosarcoma CM-induced angiogenesis in vitro and in vivo. CCL5 knockdown dramatically abolished tumor growth and angiogenesis in the osteosarcoma xenograft animal model. Importantly, we demonstrated that the expression of CCL5 and VEGF were correlated with tumor stage according the immunohistochemistry analysis of human osteosarcoma tissues. Taken together, our findings provide evidence that CCL5/CCR5 axis promotes VEGF-dependent tumor angiogenesis in human osteosarcoma microenvironment through PKCδ/c-Src/HIF-1α signaling pathway. CCL5 may represent a potential therapeutic target against human osteosarcoma.
authors.affiliation: Department of Medicine, Mackay Medical College, New Taipei City 252, Taiwan.
authors.forename: Shih-Wei
authors.initials: SW
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics, Mackay Memorial Hospital, Taipei 104, Taiwan.
authors.forename: Shih-Chia
authors.initials: SC
authors.lastname: Liu
authors.affiliation: Department of Molecular Virology, Immunology and Medical Genetics, Ohio state University, Columbus, OH 43210, USA.
authors.forename: Hui-Lung
authors.initials: HL
authors.lastname: Sun
authors.affiliation: Department of Orthopaedics, Mackay Memorial Hospital, Taipei 104, Taiwan.
authors.forename: Te-Yang
authors.initials: TY
authors.lastname: Huang
authors.affiliation: Department of Orthopaedics, Mackay Memorial Hospital, Taipei 104, Taiwan.
authors.forename: Chia-Han
authors.initials: CH
authors.lastname: Chan
authors.affiliation: Department of Orthopaedics, Mackay Memorial Hospital, Taipei 104, Taiwan.
authors.forename: Chen-Yu
authors.initials: CY
authors.lastname: Yang
authors.affiliation: Department of Medicine, Mackay Medical College, New Taipei City 252, Taiwan, Department of Internal Medicine, Mackay Memorial Hospital, Taipei 104, Taiwan.
authors.forename: Hung-I
authors.initials: HI
authors.lastname: Yeh
authors.affiliation: Department of Biotechnology, College of Health Science, Asia University, Taichung 413, Taiwan.
authors.forename: Yuan-Li
authors.initials: YL
authors.lastname: Huang
authors.affiliation: Department of Orthopedic Surgery, Kaohsiung Chang Gung Memorial Hospital Medical Center, Kaohsiung 833, Taiwan.
authors.forename: Wen-Yi
authors.initials: WY
authors.lastname: Chou
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan, Department of Orthopedic Surgery, Taichung Veterans General Hospital, Taichung 407, Taiwan, chtang@mail.cmu.edu.tw ymlin@vghtc.gov.tw.
authors.forename: Yu-Min
authors.initials: YM
authors.lastname: Lin
authors.affiliation: Department of Biotechnology, College of Health Science, Asia University, Taichung 413, Taiwan, Graduate Institute of Basic Medical Science, China Medical University, Taichung 404, Taiwan and Department of Pharmacology, School of Medicine, China Medical University, Taichung 404, Taiwan chtang@mail.cmu.edu.tw.
authors.forename: Chih-Hsin
authors.initials: CH
authors.lastname: Tang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D002465:Cell Movement; D049109:Cell Proliferation; D018946:Chemokine CCL5 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000378:metabolism / Q000473:pathology; D047369:Chromatin Immunoprecipitation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D012516:Osteosarcoma / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D019713:Receptors, CCR5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046888:Tissue Array Analysis; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
chemical_list: C483024:CCL5 protein, human; C584933:CCR5 protein, human; D018946:Chemokine CCL5; D017077:Culture Media, Conditioned; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019713:Receptors, CCR5; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A