| Summary: | 碩士 === 慈濟大學 === 醫學生物技術研究所 === 98 === Androgen receptor (AR) plays an important role in the function of several organs including primary and accessory sexual organs, skeletal muscle, and bone, making it a desirable therapeutic target. AR acts as a ligand-regulated, DNA-binding transcription factor to regulate gene expression by binding the androgen response element (ARE) within target genes. AR was shown to affect osteogenic differentiation. However, the detailed molecular mechanisms of AR in regulating bone cell differentiation remain elusive. In this present study, we determined the role and molecular mechanism of AR in D1 cell (multipotent bone marrow stromal precursor cells) osteogenic differentiation.
The effect of androgen/AR signaling on D1 cell osteogenic differentiation was examined by ARS staining, flowcytometry - surface marker, PI staining and XTT for cell morphology, cell cycle, cell proliferation and apoptosis. We examined the expression levels of AR during the osteogenic differentiation of D1 cells by semi-quantitative RT-PCR, Western blot assay and Real Time-PCR. Dual-luciferase reporter assay was also performed to determine the transcriptional activity of AR during the osteogenic differentiation of D1 cells. The transcription factor Runx2/Cbfa1 and osteocalcin are known to be essential for multipotent mesenchymal cells to differentiate into osteoblasts. Therefore, we investigated further on the effects of AR on the expression of Cbfa-1 and osteocalcin by semi-quantitative RT-PCR and Real Time-PCR. We demonstrated that Cbfa-1 and osteocalcin mRNA level were increased on osteogenic differentiation of D1 cells by androgen treatment.
Our findings suggest that androgen/AR signaling plays a critical role in determining the fate of mesenchymal progenitor cells by affecting the cell biology and gene expression.
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