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

Autor: Slawomir Pikula, Agnieszka Strzelecka-Kiliszek, René Buchet, Joanna Bandorowicz-Pikula, Saida Mebarek, Lukasz Bozycki, Marta Romiszewska
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0301 basic medicine
Saos-2 cells
Annexins
Cell Survival
Fluorescent Antibody Technique
Bone Neoplasms
Mineralization (biology)
Proto-Oncogene Mas
Catalysis
Article
matrix vesicles
lcsh:Chemistry
Inorganic Chemistry
Extracellular matrix
03 medical and health sciences
0302 clinical medicine
Calcification
Physiologic
Cell Line
Tumor
ROCK
Humans
mineralization
Physical and Theoretical Chemistry
lcsh:QH301-705.5
Molecular Biology
Spectroscopy
Cell Proliferation
Osteosarcoma
rho-Associated Kinases
Kinase
Chemistry
Organic Chemistry
annexin A6
General Medicine
Ascorbic acid
Computer Science Applications
Cell biology
Extracellular Matrix
Vesicular transport protein
030104 developmental biology
src-Family Kinases
lcsh:Biology (General)
lcsh:QD1-999
030220 oncology & carcinogenesis
Alkaline phosphatase
Src kinase
Biomarkers
Proto-oncogene tyrosine-protein kinase Src
Zdroj: International Journal of Molecular Sciences
Volume 20
Issue 12
International Journal of Molecular Sciences, Vol 20, Iss 12, p 2872 (2019)
ISSN: 1422-0067
Popis: 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 &mu
g/mL of ascorbic acid (AA) and 7.5 mM of &beta
glycerophosphate (&beta
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.
Databáze: OpenAIRE
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