Article
miR-126 regulates platelet-derived growth factor receptor-α expression and migration of primary human osteoblasts
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Authors
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Filip Simunovic
- Universitätsklinikum Freiburg, Plastische und Hand Chirurgie, Freiburg, Deutschland
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Yvonne Schmidt
- Universitätsklinikum Freiburg, Plastische und Hand Chirurgie, Freiburg, Deutschland
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Sandra Strassburg
- Universitätsklinikum Freiburg, Plastische und Hand Chirurgie, Freiburg, Deutschland
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Dietmar Pfeifer
- Universitätsklinikum Freiburg, Klinik für Innere Medizin I, Freiburg, Deutschland
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G. Björn Stark
- Universitätsklinikum Freiburg, Plastische und Hand Chirurgie, Freiburg, Deutschland
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Günter Finkenzeller
- Universitätsklinikum Freiburg, Plastische und Hand Chirurgie, Freiburg, Deutschland
| Published: | April 24, 2015 |
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Outline
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Introduction: One of the strategies to improve bone tissue engineering is to coimplant osteoblasts (hOB) and endothelial cells (HUVEC). Investigating the direct coculture of these cells in vitro, we previously showed a downregulation of platelet-derived growth factor receptor-α (PDGFR-α) in hOBs, which was a consequence of reduced mRNA stability. In the current study we investigated the possible involvement of microRNAs in this process.
Material and methods: We performed a microarray analysis of osteoblastic miRNAs following cocultivation with HUVECs. The results were verified by RT-qPCR. miRNA mimics and inhibitors were used to investigate the functional role of miRNA-126. A PDGFR-α blocking antibody was used inhibit PDGFR-α. Cellular migration was observed in the scratch migration assay. Cell proliferation was measured by cell counting, apoptosis by cell-death ELISA and osteoblast differentiation by van Kossa staining and Ca2 -quantification.
Results: The microarray revealed a marked upregulation of miR-126 in hOBs after cocultivation mit HUVECs. This result was confirmed by RT-qPCR, and it was shown that the increase is dependent on direct cell-to-cell contact. Gain-of-function and loss-of-function experiments showed that miR-126 is a negative regulator of PDGFR-α mRNA. Additionally, migration of hOBs was inhibited by miR-126 overexpression and stimulated by miR-126 inhibition. Addition of PDGFR-α blocking antibody to hOB culture also inhibited hOB migration. There was no effect of miR-126 modulation on osteoblast proliferation, apoptosis rate or differentiation.
Conclusion: This data suggest that the miR-126/PDGFR-α system regulates the migratory behavior of human osteoblasts, without exerting effects on cell survival and differentiation.
