Effects of phosphates on the expression of tissue-nonspecific alkaline phosphatase gene and phosphate-regulating genes in short-term cultures of human osteosarcoma cell lines

Orimo, Hideo; Shimada, Takashi

doi:10.1007/s11010-006-1520-6

Cited by 21 publications

(5 citation statements)

References 45 publications

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“…SaOS-2 cells have been reported to be much more tumorigenic and metastatic with respect to U-2 OS (50). Moreover, SaOS-2 cells may mimic undifferentiated osteoblasts, while MG-63 cells may represent a more differentiated stage (51). This has been also proven by proteomics, indicating that SaOS-2 express proteins typical of immature osteoblasts (24).…”

Section: Si-83 Pyrazolo[34-d]pyrimidine Inhibits Cell Viability In Fmentioning

confidence: 70%

Antiproliferative and proapoptotic activities of new pyrazolo[3,4‐d]pyrimidine derivative Src kinase inhibitors in human osteosarcoma cells

et al. 2008

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Osteosarcoma is the most frequent primitive malignant tumor of the skeletal system, characterized by an extremely aggressive clinical course that still lacks an effective treatment. Src kinase seems to be involved in the osteosarcoma malignant phenotype. We show that the treatment of human osteosarcoma cell lines with a new pyrazolo[3,4-d]pyrimidine derivative Src inhibitor, namely SI-83, impaired cell viability, with a half-maximal inhibitory concentration of 12 microM in nonstarved cells and a kinetic different from that known for the Src inhibitor PP2. Analysis by terminal deoxynucleotidyl transferase-mediated nick end labeling, Hoechst, and flow cytometric assay showed that SI-83 induced apoptosis in SaOS-2 cells. Moreover, SI-83, by inhibiting Src phosphorylation, decreased in vivo osteosarcoma tumor mass in a mouse model. Finally, SI-83 showed selectivity for osteosarcoma, since it had a far lower effect in primary human osteoblasts. These results show that human osteosarcoma had Src-dependent proliferation and that modulation of Src activity may be a therapeutic target of this new compound with low toxicity for nonneoplastic cells.

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Section: Si-83 Pyrazolo[34-d]pyrimidine Inhibits Cell Viability In Fmentioning

confidence: 70%

Antiproliferative and proapoptotic activities of new pyrazolo[3,4‐d]pyrimidine derivative Src kinase inhibitors in human osteosarcoma cells

et al. 2008

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“…Addition of NaH 2 PO 4 or β-glycerophosphate into a human osteosarcoma cell line (SaOS-2) resulted in the induction ALP mRNA expression and enzymatic activity [5, 29]. In addition, glycerol, one of the products of ALP induced hydrolysis of β-glycerophosphate, had no effect on ALP activity [5].…”

Section: Discussionmentioning

confidence: 99%

Immobilization of alkaline phosphatase on microporous nanofibrous fibrin scaffolds for bone tissue engineering

2009

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Alkaline phosphatase (ALP) promotes bone formation by degrading inorganic pyrophosphate (PP i ), an inhibitor of hydroxyapatite formation, and generating inorganic phosphate (P i ), an inducer of hydroxyapatite formation. P i is a crucial molecule in differentiation and mineralization of osteoblasts. In this study, a method to immobilize ALP on fibrin scaffolds with tightly controllable pore size and pore interconnection was developed, and the biological properties of these scaffolds were characterized both in vitro and in vivo. Microporous, nanofibrous fibrin scaffolds (FS) were fabricated using a sphere-templating method. ALP was covalently immobilized on the fibrin scaffolds using 1-ethyl-3-(dimethylaminopropyl)carbodiimide hydrochloride (EDC). Scanning electron microscopic observation (SEM) showed that mineral was deposited on immobilized alkaline phosphatase fibrin scaffolds (immobilized ALP/FS) when incubated in medium supplemented with β-glycerophosphate, suggesting that the immobilized ALP was active. Primary calvarial cells attached, spread and formed multiple layers on the surface of the scaffolds. Mineral deposition was also observed when calvarial cells were seeded on immobilized ALP/FS. Furthermore, cells seeded on immobilized ALP/FS exhibited higher osteoblast marker gene expression compared to control FS. Upon implantation in mouse calvarial defects, both the immobilized ALP/FS and FS alone treated group had higher bone volume in the defect compared to the empty defect control. Furthermore, bone formation in the immobilized ALP/FS treated group was statistically significant compared to FS alone group. However, the response was not robust.

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“…BGP can be replaced with physiologic levels of inorganic phosphate or other phosphate esters (ATP, pyridoxal phosphate, phosphoserine, etc , In addition, expression of some key genes relevant to mineralization has been shown to be comparable in mineralizing cultures with BGP or inorganic phosphate. ,, Thus, authors and readers alike should remember that the presence of hydroxyapatite in cultures with BGP simply indicates the presence of alkaline phosphatase or other phosphatase activity, especially since mineralization does not occur in these systems when the phosphatases are inhibited.…”

Section: Conclusion: Advice For Cell Culture Studies Of Calcificationmentioning

confidence: 99%

Cell Culture Systems for Studies of Bone and Tooth Mineralization

2008

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1. Introduction 4716 1.1. Cells and Tissues of Bones and Teeth 4716 1.1.1. The Mineral and Extracellular Matrix of Bones and Teeth 4716 1.1.2. Cells in Mineralized Tissues 4717 2. Is the Mineral Formed in Culture Similar to "Physiologic Mineral"? 4717 2.1. X-ray Diffraction 4718 2.2. SEM and TEM and Related Techniques 4718 2.3. Atomic Force Microscopy 4718 2.4. Light Microscopy 4719 2.5. Vibrational Spectroscopy and Vibrational Spectroscopic Imaging 4719 2.6. Radiographic and Related Methods 4720 2.6.1. Magnetic Resonance Methods 4720 2.6.2. Microcomputed Tomography (microCT) or X-ray Microtomography (XMT) 4720 2.7. Chemical Analyses 4721 3. Systems for Studying Mineralization in Culture 4721 3.1. Cell Sources 4721 3.1.1. Organ Culture 4721 3.

show abstract

Effects of phosphates on the expression of tissue-nonspecific alkaline phosphatase gene and phosphate-regulating genes in short-term cultures of human osteosarcoma cell lines

Cited by 21 publications

References 45 publications

Antiproliferative and proapoptotic activities of new pyrazolo[3,4‐d]pyrimidine derivative Src kinase inhibitors in human osteosarcoma cells

Antiproliferative and proapoptotic activities of new pyrazolo[3,4‐d]pyrimidine derivative Src kinase inhibitors in human osteosarcoma cells

Immobilization of alkaline phosphatase on microporous nanofibrous fibrin scaffolds for bone tissue engineering

Cell Culture Systems for Studies of Bone and Tooth Mineralization

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