Thrombin induces proliferation of osteoblast-like cells through phosphatidylcholine hydrolysis

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

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

We examined the effect of thrombin on phosphatidylcholine-hydrolyzing phospholipase D activity in osteoblast-like MC3T3-E1 cells. Thrombin stimulated the formation of choline dose dependently in the range between 0.01 and 1 U/ml, but not the phosphocholine formation. Diisopropylfluorophosphate (DFP)-inactivated thrombin had little effect on the choline formation. The combined effects of thrombin and 12-O-tetradecanoylphorbol-13-acetate, a protein kinase C-activating phorbol ester, on the choline formation were additive. Staurosporine, an inhibitor of protein kinases, had little effect on the thrombin-induced formation of choline. Combined addition of thrombin and NaF, an activator of heterotrimeric GTP-binding protein, did not stimulate the formation of choline further. Pertussis toxin had little effect on the thrombin-induced formation of choline. Thrombin stimulated Ca 2+ influx from extracellular space time and dose dependently. The depletion of extracellular Ca 2+ by EGTA exclusively reduced the thrombin-induced choline formation. Thrombin had only a slight effect on phosphoinositide-hydrolyzing phospholipase C activity. Thrombin induced diacylglyceroI formation and DNA synthesis, and increased the number of MC3T3-E1 cells, but DFP-inactivated thrombin did not. Thrombin suppressed both basal and fetal calf serum-induced alkaline phosphatase activity in these cells. Propranolol, an inhibitor of phosphatidic acid phosphohydrolase, inhibited both the thrombin-induced diacylglycerol formation and DNA synthesis. These results suggest that thrombin stimulates phosphatidylcholine-hydrolyzing phospholipase D due to self-induced Ca 2+ influx independently of protein kinase C activation in osteoblast-like cells and that its proliferative effect depends on phospholipase D activation.

Original languageEnglish
Pages (from-to)209-216
Number of pages8
JournalJournal of Cellular Physiology
Volume168
Issue number1
DOIs
Publication statusPublished - 01-07-1996
Externally publishedYes

Fingerprint

Osteoblasts
Phosphatidylcholines
Thrombin
Hydrolysis
Choline
Phospholipase D
Isoflurophate
Protein Kinase C
Chemical activation
Phosphatidate Phosphatase
Phosphoinositide Phospholipase C
Heterotrimeric GTP-Binding Proteins
Staurosporine
Phosphorylcholine
Egtazic Acid
DNA
Diglycerides
Pertussis Toxin
Extracellular Space
Type C Phospholipases

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Suzuki, Atsushi ; Kozawa, Osamu ; Shinoda, Junji ; Watanabe, Yasuko ; Saito, Hidehiko ; Oiso, Yutaka. / Thrombin induces proliferation of osteoblast-like cells through phosphatidylcholine hydrolysis. In: Journal of Cellular Physiology. 1996 ; Vol. 168, No. 1. pp. 209-216.
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Thrombin induces proliferation of osteoblast-like cells through phosphatidylcholine hydrolysis. / Suzuki, Atsushi; Kozawa, Osamu; Shinoda, Junji; Watanabe, Yasuko; Saito, Hidehiko; Oiso, Yutaka.

In: Journal of Cellular Physiology, Vol. 168, No. 1, 01.07.1996, p. 209-216.

Research output: Contribution to journalArticle

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AU - Suzuki, Atsushi

AU - Kozawa, Osamu

AU - Shinoda, Junji

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