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

Yuan-Ya Liao,I-Ping Chiang,Shih-Wei Wang,Yu-Min Lin,Pei-Yu Chou,Shao-Keh Hsu,Chih-Hsin Tang,Tzu-Ming Chang,Hsiao-Chi Tsai,Ming-Chih Chou,Hsien-Te Chen,Yi-Chin Fong

doi:10.18632/oncotarget.6708

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.

Highlights

Angiogenesis is a physiological process through which new blood vessels form from pre-existing vessels
We found that the conditioned medium (CM) from CCL3-treated osteosarcoma cells promoted endothelial progenitor cell (EPC) migration and tube formation (VEGF-A was used as positive control) (Fig. 1E1G and Supplementary Fig. S1)
CCL3 promotes angiogenesis through Vascular endothelial growth factor-A (VEGF-A)-dependent expression in human osteosarcoma cells

Summary

Introduction

Angiogenesis is a physiological process through which new blood vessels form from pre-existing vessels. It is involved in the growth, maintenance, and metastasis of most of the solid tumors [1, 2]. The use of chemotherapy has remarkably augmented the survival rates; patients have about 65-70% chance of 5-year relapse-free survival, if the disease is localized [3]. Patients with pulmonary metastasis of osteosarcoma have a very poor prognosis with a relapse within 5 years [4, 5]. It is known that vascular endothelial growth factor-A (VEGF-A) is one of the major inducers of angiogenesis [6] and plays an important role in the pathogenesis and progression of various cancers [7]. It is necessary to understand the effect of VEGF-A in angiogenesis pathways and its mechanism in human osteosarcoma

Methods

Results

Conclusion