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Open AccessArticle

Src and ROCK Kinases Differentially Regulate Mineralization of Human Osteosarcoma Saos-2 Cells

1
Laboratory of Biochemistry of Lipids, Nencki Institute of Experimental Biology, Polish Academy of Sciences, 3 Pasteur Str., 02-093 Warsaw, Poland
2
Université de Lyon, CEDEX 69622 Villeurbanne, France
3
Université Lyon 1, CEDEX 69622 Villeurbanne, France
4
NSA de Lyon, CEDEX 69621 Villeurbanne, France
5
CPE Lyon, CEDEX 69616 Villeurbanne, France
6
ICBMS CNRS UMR 5246, CEDEX 69622 Villeurbanne, France
7
Laboratory of Cellular Metabolism, Nencki Institute of Experimental Biology, Polish Academy of Sciences, 3 Pasteur Str., 02-093 Warsaw, Poland
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(12), 2872; https://doi.org/10.3390/ijms20122872
Received: 15 April 2019 / Revised: 28 May 2019 / Accepted: 10 June 2019 / Published: 12 June 2019
(This article belongs to the Special Issue Kinase Signal Transduction 2019)
Osteoblasts initiate bone mineralization by releasing matrix vesicles (MVs) into the extracellular matrix (ECM). MVs promote the nucleation process of apatite formation from Ca2+ and Pi in their lumen and bud from the microvilli of osteoblasts during bone development. Tissue non-specific alkaline phosphatase (TNAP) as well as annexins (among them, AnxA6) are abundant proteins in MVs that are engaged in mineralization. In addition, sarcoma proto-oncogene tyrosine-protein (Src) kinase and Rho-associated coiled-coil (ROCK) kinases, which are involved in vesicular transport, may also regulate the mineralization process. Upon stimulation in osteogenic medium containing 50 μg/mL of ascorbic acid (AA) and 7.5 mM of β-glycerophosphate (β-GP), human osteosarcoma Saos-2 cells initiated mineralization, as evidenced by Alizarin Red-S (AR-S) staining, TNAP activity, and the partial translocation of AnxA6 from cytoplasm to the plasma membrane. The addition of 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo [3,4-d] pyrimidine (PP2), which is an inhibitor of Src kinase, significantly inhibited the mineralization process when evaluated by the above criteria. In contrast, the addition of (R)-(+)-trans-4-(1-aminoethyl)-N-(4-pyridyl) cyclohexane carboxamide hydrochloride (Y-27632), which is an inhibitor of ROCK kinase, did not affect significantly the mineralization induced in stimulated Saos-2 cells as denoted by AR-S and TNAP activity. In conclusion, mineralization by human osteosarcoma Saos-2 cells seems to be differently regulated by Src and ROCK kinases. View Full-Text
Keywords: Src kinase; ROCK; annexin A6; mineralization; matrix vesicles; Saos-2 cells Src kinase; ROCK; annexin A6; mineralization; matrix vesicles; Saos-2 cells
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Strzelecka-Kiliszek, A.; Romiszewska, M.; Bozycki, L.; Mebarek, S.; Bandorowicz-Pikula, J.; Buchet, R.; Pikula, S. Src and ROCK Kinases Differentially Regulate Mineralization of Human Osteosarcoma Saos-2 Cells. Int. J. Mol. Sci. 2019, 20, 2872.

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