CCL3 promotes angiogenesis by dysregulation of miR-374b/ VEGF-A axis in human osteosarcoma cells

// Yuan-Ya Liao 1, 2, * , Hsiao-Chi Tsai 3, * , Pei-Yu Chou 4 , Shih-Wei Wang 5 , Hsien-Te Chen 6, 7 , Yu-Min Lin 8, 9 , I-Ping Chiang 10 , Tzu-Ming Chang 11 , Shao-Keh Hsu 11 , Ming-Chih Chou 1, 2 , Chih-Hsin Tang 3, 12, 13 , Yi-Chin Fong 6, 14 1 Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan 2 Department of Surgery, Chung Shan Medical University Hospital, Taichung, Taiwan 3 Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan 4 Department of Nursing, Hung Kuang University, Taichung, Taiwan 5 Department of Medicine, Mackay Medical College, New Taipei City, Taiwan 6 Department of Orthopedic Surgery, China Medical University Hospital, Taichung, Taiwan 7 School of Chinese Medicine, China Medical University, Taichung, Taiwan 8 Department of Medicine, Chung Shan Medical University, Taichung, Taiwan 9 Department of Orthopaedics, Taichung Veterans General Hospital, Taichung, Taiwan 10 Department of Pathology, China Medical University Hospital, Taichung, Taiwan 11 Department of Orthopedic Surgery, Tungs' Taichung Metroharbor Hospital, Taichung, Taiwan 12 Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan 13 Department of Biotechnology, College of Health Science, Asia University, Taichung, Taiwan 14 Department of Sports Medicine, College of Health Care, China Medical University, Taichung, Taiwan * These authors have contributed equally to this work Correspondence to: Yi-Chin Fong, e-mail: yichin.fong@gmail.com Chih-Hsin Tang, e-mail: chtang@mail.cmu.edu.tw Ming-Chih Chou, e-mail: cshe032@csh.org.tw Keywords: MAPK, miR-374b, VEGF-A, angiogenesis, osteosarcoma Received: July 27, 2015 Accepted: December 05, 2015 Published: December 21, 2015 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.