The effect of combination of β-catenin and bone morphogenetic protein 9 on osteogenic differentiation of osteosarcoma cell line TE85 / 肿瘤

ABSTRACT

Objective: To investigate the effect of β-catenin and bone morphogenetic protein 9 (BMP-9) on osteogenic differentiation of osteosarcoma TE85 cells. Methods: The TE85 cells were infected by recombinant adenovirus Adβ-catenin and AdBMP9 alone or in combination. The expression levels of β-catenin and BMP9 mRNAs in TE85 cells were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The relative β-catenin/TCF4 activity was detected by Luciferase Reported System. Alkaline phosphatase (ALP) staining and numeration were used to detect the early osteogenic potential of TE85 cells, and the alizarin red staining was performed to detect the late osteogenic potential. The expression levels of osteoprotegerin (OPG), osteocalcin (OC) and osteopontin (OPN) mRNAs were detected by semi-quantitative RT-PCR, and the expression levels of OC and OPN proteins were detected by Western blotting. Results: The exogenous mRNAs of β-catenin and BMP9 in TE85 cells were increased by adenovirus-mediated Adβ-catenin and AdBMP9 (both P < 0.05), and the relative β-catenin/TCF4 activity was increased after infection with Adβ-catenin (P < 0.05). Adβ-catenin or AdBMP9 infection could enhance the activity of ALP in a limited range in TE85 cells (P < 0.05), but they could not promote the mRNA and protein expressions of late osteogenic biomarkers OPN and OC as well as the calcium deposition. However, both early and late potential osteogenic abilities were enhanced obviously after infection with combination of Adβ-catenin and AdBMP9 (all P < 0.05). Conclusion: Osteogenic differentiation can be limitedly induced by β-catenin and BMP9 alone in TE85 cells, but this differentiation ability can be remarkably enhanced by combination of β-catenin and BMP9 with a synergistic effect.