BMPR2 promotes invasion and metastasis via the RhoA-ROCK-LIMK2 pathway in human osteosarcoma cells.

Tingting Ren,Wei Guo,Shidong Wang,Yi Huang,Bingxin Zheng,Guangjun Jiao,Kuisheng Liu,Xing Bao,Kunkun Sun,Fan Zhang

doi:10.18632/oncotarget.17382

Abstract

Bone morphogenetic protein receptor 2 (BMPR2) has been identified in several types of cancer. However, its role in osteosarcoma is largely unknown. We systematically investigated the role of BMPR2 in osteosarcoma cell lines, human tissue samples and xenograft models. The relationship between BMPR2 expression and osteosarcoma patients’ survival was investigated by bioinformatics and clinical data. Wound healing assay and transwell assay were used to detect the changes of cell migration and invasion ability after BMPR2 transfection. In addition, downstream phosphoproteins were analyzed by iTRAQ-based phosphoproteomic analysis and verified by western blotting. In vivo, the effects of BMPR2 on the growth, formation and metastasis of 143B cells were observed by orthotopic transplantation of nude mice. Here, we demonstrated that BMPR2 expression was elevated in a majority of osteosarcoma tissues compared with normal bone tissue. Osteosarcoma patients with greater BMPR2 expressing level showed a poor overall survival. The depletion of BMPR2 in 143B cells markedly reduced the invasive capacity in vitro and metastatic potential in vivo. Mechanistically, we found that LIM domain kinase 2 (LIMK2) was phosphorylated and activated by BMPR2 and that this event was crucial for activation of the BMPR2-mediated signal pathway in osteosarcoma cells. Additionally, we demonstrated that BMPR2 could active LIMK2 through the RhoA/ROCK pathway and could also interact with LIMK2 directly. Taken together, our study revealed that BMPR2 functions as a prometastatic oncogene in vitro and in vivo with the activation of the RhoA-ROCK-LIMK2 pathway and may represent a potential therapeutic target for osteosarcoma.

Highlights

Osteosarcoma is the most common primary bone malignancy diagnosed in children and adolescents and has a high propensity for local invasion and distant metastasis [1,2,3,4]
To investigate the relationship between Bone morphogenetic protein receptor 2 (BMPR2) levels and the outcomes in osteosarcoma, we first searched for publically available datasets. 88 cases in the GSE33383 dataset were able to be involved in the survival analysis, which indicated high BMPR2 expression may be implicated with decreased metastasis-free survival and overall survival in osteosarcoma (Figure 1A)
Immunohistochemistry detection of BMPR2 was performed on 67 osteosarcoma samples, which was divided into a high and low expression group according to the cutoff value (Figure 1C)

Summary

Introduction

Osteosarcoma is the most common primary bone malignancy diagnosed in children and adolescents and has a high propensity for local invasion and distant metastasis [1,2,3,4]. Advances of neoadjuvant chemotherapy and the refinements of surgical techniques have greatly raised the 5-year survival rate of patients with localized osteosarcoma. Bone morphogenetic proteins (BMPs) are multifunctional proteins that are members of the transforming growth factor β (TGF-β) superfamily [5, 6]. They play an important role during development and regulate many processes, including cellular proliferation, adhesion, migration, differentiation, inflammation, and apoptosis [7,8,9]. BMP signaling can follow a Smad-independent pathway through several intracellular mediators such as p38 MAPK, ERK and JNK [5]

Methods

Results

Conclusion