title: CCL3 promotes angiogenesis by dysregulation of miR-374b/ VEGF-A axis in human osteosarcoma cells.
abstract: Osteosarcoma is the most frequent bone tumor, characterized by a high metastatic potential. However, the crosstalk between chemokine (C-C motif) ligand 3 (CCL3), which facilitates tumor progression and metastasis. Vascular endothelial growth factor-A (VEGF-A), an angiogenesis inducer and a highly specific mitogen for endothelial cells, has not been well explored in human osteosarcoma. Here we demonstrate the correlation of CCL3 and VEGF-A expressions, quantified by immunohistochemistry, with the tumor stage of human osteosarcoma tissues. Furthermore, CCL3 promotes VEGF-A expression in human osteosarcoma cells that subsequently induces human endothelial progenitor cell (EPC) migration and tube formation. Phosphorylation of JNK, ERK, and p38 was found after CCL3 stimulation. In addition, JNK, ERK, and p38 inhibitors also abolished CCL3-induced VEGF-A expression and angiogenesis. We noted that CCL3 reduces the expression of miR-374b and miR-374b mimic by reversing CCL3-promoted VEGF-A expression and angiogenesis in vitro and in vivo. This study shows that CCL3 promotes VEGF-A expression and angiogenesis in human osteosarcoma cells by down-regulating miR-374b expression via JNK, ERK, and p38 signaling pathways. Thus, CCL3 may be a new molecular therapeutic target in osteosarcoma angiogenesis and metastasis.
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.|Department of Surgery, Chung Shan Medical University Hospital, Taichung, Taiwan.
authors.forename: Yuan-Ya
authors.initials: YY
authors.lastname: Liao
authors.affiliation: Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan.
authors.forename: Hsiao-Chi
authors.initials: HC
authors.lastname: Tsai
authors.affiliation: Department of Nursing, Hung Kuang University, Taichung, Taiwan.
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Chou
authors.affiliation: Department of Medicine, Mackay Medical College, New Taipei City, Taiwan.
authors.forename: Shih-Wei
authors.initials: SW
authors.lastname: Wang
authors.affiliation: Department of Orthopedic Surgery, China Medical University Hospital, Taichung, Taiwan.|School of Chinese Medicine, China Medical University, Taichung, Taiwan.
authors.forename: Hsien-Te
authors.initials: HT
authors.lastname: Chen
authors.affiliation: Department of Medicine, Chung Shan Medical University, Taichung, Taiwan.|Department of Orthopaedics, Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Yu-Min
authors.initials: YM
authors.lastname: Lin
authors.affiliation: Department of Pathology, China Medical University Hospital, Taichung, Taiwan.
authors.forename: I-Ping
authors.initials: IP
authors.lastname: Chiang
authors.affiliation: Department of Orthopedic Surgery, Tungs' Taichung Metroharbor Hospital, Taichung, Taiwan.
authors.forename: Tzu-Ming
authors.initials: TM
authors.lastname: Chang
authors.affiliation: Department of Orthopedic Surgery, Tungs' Taichung Metroharbor Hospital, Taichung, Taiwan.
authors.forename: Shao-Keh
authors.initials: SK
authors.lastname: Hsu
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.|Department of Surgery, Chung Shan Medical University Hospital, Taichung, Taiwan.
authors.forename: Ming-Chih
authors.initials: MC
authors.lastname: Chou
authors.affiliation: Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan.|Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan.|Department of Biotechnology, College of Health Science, Asia University, Taichung, Taiwan.
authors.forename: Chih-Hsin
authors.initials: CH
authors.lastname: Tang
authors.affiliation: Department of Orthopedic Surgery, China Medical University Hospital, Taichung, Taiwan.|Department of Sports Medicine, College of Health Care, China Medical University, Taichung, Taiwan.
authors.forename: Yi-Chin
authors.initials: YC
authors.lastname: Fong
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D001859:Bone Neoplasms / Q000098:blood supply* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D054405:Chemokine CCL3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D049033:Chorioallantoic Membrane; D066026:Endothelial Progenitor Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D009389:Neovascularization, Pathologic / Q000473:pathology*; D012516:Osteosarcoma / Q000098:blood supply* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
chemical_list: C515429:CCL3 protein, human; D054405:Chemokine CCL3; C587995:MIRN374 microRNA 374, human; D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: MAPK; VEGF-A; angiogenesis; miR-374b; osteosarcoma