Msx2 plays an important role in BMP6-induced osteogenic differentiation of two mesenchymal cell lines: C3H10T1/2 and C2C12

Chuan Cai; Jing Wang; Na Huo; Li Wen; Peng Xue; Ye Huang

doi:10.1016/j.reth.2020.03.015

Msx2 plays an important role in BMP6-induced osteogenic differentiation of two mesenchymal cell lines: C3H10T1/2 and C2C12

Regen Ther. 2020 May 15:14:245-251. doi: 10.1016/j.reth.2020.03.015. eCollection 2020 Jun.

Authors

Chuan Cai¹, Jing Wang¹, Na Huo¹, Li Wen¹, Peng Xue¹, Ye Huang²

Affiliations

¹ Department of Stomatology, The First Medical Center, Chinese PLA General Hospital, Beijing, 100853, China.
² Department of Dermatology, Air Force General Hospital of Chinese PLA, Beijing, 100412, China.

Abstract

Bone morphogenetic proteins (BMPs), have been shown to enhance the osteogenic differentiation of mesenchymal cells (MCs) and to promote bone formation. BMP6 is known to play an important role in the process of MCs towards osteogenic differentiation by virtue of their osteoinductive and cell type specific proliferative activity. However, the molecular mechanism relate to BMP6 osteoinductive activity is still unclear and continues to warrant further investigation. Msx2 is a member of the homeobox gene family of transcription factors and promotes calcification. Hence, we wondered if it might also play a role in BMP6-induced osteogenesis. In this study, two mouse mesenchymal cell lines were treated with BMP6, adenovirus-Msx2 (Ad-Msx2) or adenovirus-siMsx2 (Ad-siMsx2). Based on the results of mRNA and protein expression, it was indicated that BMP6 could enhance the expression of Msx2 and activate the phosphorylation of Smad 1/5/8, p38 and ERK1/2. Being transfected by Ad-Msx2, the BMP6-induced activation of phosphorylation was significantly promoted. On the contrary, two cell lines transfected by Ad-siMsx2 presented an inhibited expression of three phosphorylated proteins even after being induced by BMP6. The evaluation of ALP, OPN, OC and calcium deposits revealed the osteogenic results those were corresponding to the results of mRNA and protein. Taken together, these findings can be a novel viewpoint for the understanding of the mechanisms of BMP6-induced osteogenesis and provide therapeutic targets of bone defect.

Keywords: Ad, adenovirus-transfection; Adenovirus-transfection; BMP, Bone morphogenetic protein; BMP6; Msx, Msh homeobox; Msx2; Osteogenesis; Signaling pathway; siRNA, silencing RNA.