The role of BMP6 in the proliferation and differentiation of chicken cartilage cells.

Fei Ye,Qing Zhu,Diyan Li,Xiaoling Zhao,Yan Wang,Hengyong Xu,Huadong Yin,Wei Shi

doi:10.1371/journal.pone.0204384

Abstract

Previous studies have indicated that bone morphogenetic protein (BMP) 6 may play an important role in skeletal system development and progression. However, the mechanism underlying the effects of BMP6 in cartilage cell proliferation and differentiation remains unknown. In this study, cartilage cells were isolated from shanks of chicken embryos and treated with different concentrations of Growth Hormone. Cell proliferation potential was assessed using real-time polymerase chain reaction (RT-PCR), western blotting and CCK-8 assays in vitro. The results showed that at 48 h, the Collagen II and BMP6 expression levels in 50 ng/μl GH-treated cartilage cells were significantly higher than in groups treated with 100 ng/μl or 200 ng/μl GH. We further observed that knockdown of BMP6 in cartilage cells led to significantly decreased expression mRNAs and proteins of Collagen II and Collagen X. Moreover, the suppression of BMP6 expression by a specific siRNA led to significantly decreased expression mRNA levels of IGF1R, JAK2, PKC, PTH, IHH and PTHrP and decreased protein levels of PKC, IHH and PTHrP. Taken together, our data suggest that BMP6 may play a critical role in chicken cartilage cell proliferation and differentiation through the regulation of IGF1, JAK2, PKC, PTH, and IHH-PTHrP signaling pathways.

Highlights

Bone morphogenetic proteins (BMPs) are secreted-type multifunctional proteins belonging to the transforming growth factor (TGF)-β superfamily
Local injection of GH can increase the number of cartilage cells in rats [18].In this study, we first extracted and cultured cartilage cells from different breeds of chickens, and we investigated the expression of BMP6 and the changes in expression of key genes involved in related signaling pathways through GH (Growth hormone)-mediated induction at different concentrations to determine its potential role in cell proliferation and differentiation
In our previous studies we found that the polymorphism in BMP6 gene in chicken had significant correlation with femur perimeter [32], but the expression of BMP6 in cartilage plate of 15 E, 1 day avian broiler and yellow bantam chickens detected by real-time polymerase chain reaction (RT-PCR) was high

Summary

Introduction

Bone morphogenetic proteins (BMPs) are secreted-type multifunctional proteins belonging to the transforming growth factor (TGF)-β superfamily. Many studies have reported that BMPs play very important roles in bone formation and cartilage induction in both vertebrates and invertebrates [1,2]; they are considered crucial molecules involved in cell growth, differentiation, chemotaxis and apoptosis during embryonic development and postnatal tissue remodeling [3]. BMPs stimulate target cells mainly through their specific type I and type II receptors on the cell membrane. BMPs usually combine with the type II receptor, activation of receptor type I [4,5]. BMPs first bind to the receptors on the membrane and transmit this signal through the Smads pathway to promote the differentiation of chondrocytes into the osteogenic lineage [6]. In addition to the Smads signaling pathway, other signaling pathways can be transmitted from BMP family, such as mitogen-activated protein kinase (MAPK) pathways [7,8]

Results

Discussion

Conclusion