Effect of platelet-derived growth factor on phosphatidylcholine-hydrolyzing phospholipase d in osteoblast-like cells

Osamu Kozawa, Atsushi Suzuki, Yasuko Watanabe, Junji Shinoda, Yutaka Oiso

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Abstract

We examined the effect of platelet-derived growth factor (PDGF) on the activation of phosphatidylcholine-hydrolyzing phospholipase D in osteoblast-like MC3T3-E1 cells. PDGF-BB stimulated both the formation of choline (EC50 = 15 ng/ml) and inositol phosphates (EC50 = 5 ng/ml). However, PDGF-BB had little effect on the formation of phosphocholine. The formation of choline stimulated by a combination of PDGF-BB and 12-O-tetradecanoylphorbol-13-acetate, a protein kinase C (PKC)-activating phorbol ester, was additive. H-7, an inhibitor of protein kinases, inhibited 12-O-tetradecanoylphorbol-13-acetate-induced choline formation, whereas HA1004, a control for H-7 as PKC inhibitor, had little effect. Neither H-7 nor HA1004 affected the PDGF-BB-induced formation of choline. Genistein and methyl 2,5-dihydroxycinnamate, inhibitors of protein tyrosine kinases, dose dependently inhibited the PDGF-BB-induced formation of choline. PDGF-BB stimulated Ca2+ influx from extracellular space. PDGF-BB-induced choline formation was significantly reduced by chelating extracellular Ca2+ with EGTA. PDGF-BB stimulated DNA synthesis of MC3YT3-E1 cells, and H-7 inhibited the DNA synthesis. These results strongly suggest that PDGF activates phosphatidyl-choline-hydrolyzing phospholipase D independently from PKC activated by phosphoinositide hydrolysis in osteoblast-like cells, and that both tyrosine kinase activation and Ca2+ influx are essential for this mechanism.

Original languageEnglish
Pages (from-to)4473-4478
Number of pages6
JournalEndocrinology
Volume136
Issue number10
DOIs
Publication statusPublished - 10-1995

Fingerprint

Phospholipases
Platelet-Derived Growth Factor
Osteoblasts
Phosphatidylcholines
Choline
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
Protein Kinase C
Phospholipase D
Tetradecanoylphorbol Acetate
Protein Kinase Inhibitors
Protein-Tyrosine Kinases
Inositol Phosphates
Phosphorylcholine
platelet-derived growth factor BB
Protein C Inhibitor
Genistein
Egtazic Acid
DNA
Extracellular Space
Phorbol Esters

All Science Journal Classification (ASJC) codes

  • Endocrinology

Cite this

Kozawa, Osamu ; Suzuki, Atsushi ; Watanabe, Yasuko ; Shinoda, Junji ; Oiso, Yutaka. / Effect of platelet-derived growth factor on phosphatidylcholine-hydrolyzing phospholipase d in osteoblast-like cells. In: Endocrinology. 1995 ; Vol. 136, No. 10. pp. 4473-4478.
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Effect of platelet-derived growth factor on phosphatidylcholine-hydrolyzing phospholipase d in osteoblast-like cells. / Kozawa, Osamu; Suzuki, Atsushi; Watanabe, Yasuko; Shinoda, Junji; Oiso, Yutaka.

In: Endocrinology, Vol. 136, No. 10, 10.1995, p. 4473-4478.

Research output: Contribution to journalArticle

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T1 - Effect of platelet-derived growth factor on phosphatidylcholine-hydrolyzing phospholipase d in osteoblast-like cells

AU - Kozawa, Osamu

AU - Suzuki, Atsushi

AU - Watanabe, Yasuko

AU - Shinoda, Junji

AU - Oiso, Yutaka

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N2 - We examined the effect of platelet-derived growth factor (PDGF) on the activation of phosphatidylcholine-hydrolyzing phospholipase D in osteoblast-like MC3T3-E1 cells. PDGF-BB stimulated both the formation of choline (EC50 = 15 ng/ml) and inositol phosphates (EC50 = 5 ng/ml). However, PDGF-BB had little effect on the formation of phosphocholine. The formation of choline stimulated by a combination of PDGF-BB and 12-O-tetradecanoylphorbol-13-acetate, a protein kinase C (PKC)-activating phorbol ester, was additive. H-7, an inhibitor of protein kinases, inhibited 12-O-tetradecanoylphorbol-13-acetate-induced choline formation, whereas HA1004, a control for H-7 as PKC inhibitor, had little effect. Neither H-7 nor HA1004 affected the PDGF-BB-induced formation of choline. Genistein and methyl 2,5-dihydroxycinnamate, inhibitors of protein tyrosine kinases, dose dependently inhibited the PDGF-BB-induced formation of choline. PDGF-BB stimulated Ca2+ influx from extracellular space. PDGF-BB-induced choline formation was significantly reduced by chelating extracellular Ca2+ with EGTA. PDGF-BB stimulated DNA synthesis of MC3YT3-E1 cells, and H-7 inhibited the DNA synthesis. These results strongly suggest that PDGF activates phosphatidyl-choline-hydrolyzing phospholipase D independently from PKC activated by phosphoinositide hydrolysis in osteoblast-like cells, and that both tyrosine kinase activation and Ca2+ influx are essential for this mechanism.

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