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Involvement of phosphatidylcholine hydrolysis by phospholipase C in prostaglandin F2alpha-induced 1,2-diacylglycerol formation in osteoblast-like MC3T3-E1 cells.
J Bone Miner Metab. 2004; 22(3):198-206.JB

Abstract

We previously demonstrated that a prostaglandin F2alpha (PGF2alpha)-induced, sustained increase in 1,2-diacylglycerol (DAG) production was important for proliferation in osteoblast-like MC3T3-E1 cells. The 1,2-DAG formation is mediated by various enzymes, such as phos-phoinositide (PI)-specific phospholipase C (PLC), phospholipase D (PLD), and phosphatidylcholine (PC)-specific phospholipase C (PC-PLC). In the present study, to elucidate the mechanism of the 1,2-DAG formation, we have examined the PGF2alpha-induced production of [(3)H]phosphorylcholine, a product of PC-PLC activity, in [(3)H]choline-labeled MC3T3-E1 cells. The PGF2alpha-induced [(3)H]phosphorylcholine production was inhibited by genistein, a potent protein tyrosine kinase inhibitor, and increased by vanadate, a potent protein tyrosine phosphatase inhibitor. However, there were no effects after treatment with protein kinase C (PKC) inhibitors, the guanosine triphosphate (GTP) binding protein activator, NaF/AlCl(3), a Ca(2+)-ionophore, or the potent activator of PKC, phorbol 12-myristate 13-acetate (PMA), suggesting that a tyrosine kinase(s) was involved in the PGF2alpha-induced [(3)H]phosphorylcholine formation. Furthermore, a PGF2alpha analogue, 16-(3-trifluoromethylphenoxy)-Omega-tetranor-trans-Delta(2) PGF2alpha methyl ester (ONO-995), stimulated the proliferation of MC3T3-E1 cells to a level similar to that seen with PGF2alpha, and also caused phosphorylcholine and 1,2-DAG generation. However, neither an increase in intracellular free calcium ion ([Ca(2+)]i) levels by PI-PLC, nor phosphatidylethanol formation (and choline production) by PC-PLD were observed. From these results, we conclude that PGF2alpha-induced 1,2-DAG accumulation was mediated mainly via tyrosine kinase(s)-dependent PC hydrolysis by PLC activity in osteoblast-like MC3T3-E1 cells.

Authors+Show Affiliations

Sakai T
Department of Oral and Maxillo-Facial Surgery, Toho University School of Medicine, Tokyo, Japan. tsakai-gif@umin.ac.jp
Sugiyama T
No affiliation info available
Banno Y
No affiliation info available
Kato Y
No affiliation info available
Nozawa Y
No affiliation info available

MeSH

3T3 CellsAnimalsCalciumCell LineDiglyceridesDinoprostGTP-Binding ProteinsHydrolysisMiceOsteoblastsPhosphatidylcholinesProtein KinasesThymidineType C Phospholipases

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15108061

Citation

Sakai, Takayuki, et al. "Involvement of Phosphatidylcholine Hydrolysis By Phospholipase C in Prostaglandin F2alpha-induced 1,2-diacylglycerol Formation in Osteoblast-like MC3T3-E1 Cells." Journal of Bone and Mineral Metabolism, vol. 22, no. 3, 2004, pp. 198-206.
Sakai T, Sugiyama T, Banno Y, et al. Involvement of phosphatidylcholine hydrolysis by phospholipase C in prostaglandin F2alpha-induced 1,2-diacylglycerol formation in osteoblast-like MC3T3-E1 cells. J Bone Miner Metab. 2004;22(3):198-206.
Sakai, T., Sugiyama, T., Banno, Y., Kato, Y., & Nozawa, Y. (2004). Involvement of phosphatidylcholine hydrolysis by phospholipase C in prostaglandin F2alpha-induced 1,2-diacylglycerol formation in osteoblast-like MC3T3-E1 cells. Journal of Bone and Mineral Metabolism, 22(3), 198-206.
Sakai T, et al. Involvement of Phosphatidylcholine Hydrolysis By Phospholipase C in Prostaglandin F2alpha-induced 1,2-diacylglycerol Formation in Osteoblast-like MC3T3-E1 Cells. J Bone Miner Metab. 2004;22(3):198-206. PubMed PMID: 15108061.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Involvement of phosphatidylcholine hydrolysis by phospholipase C in prostaglandin F2alpha-induced 1,2-diacylglycerol formation in osteoblast-like MC3T3-E1 cells. AU - Sakai,Takayuki, AU - Sugiyama,Takatoshi, AU - Banno,Yoshiko, AU - Kato,Yukihiro, AU - Nozawa,Yoshinori, PY - 2002/08/07/received PY - 2003/07/18/accepted PY - 2004/4/27/pubmed PY - 2005/1/12/medline PY - 2004/4/27/entrez SP - 198 EP - 206 JF - Journal of bone and mineral metabolism JO - J Bone Miner Metab VL - 22 IS - 3 N2 - We previously demonstrated that a prostaglandin F2alpha (PGF2alpha)-induced, sustained increase in 1,2-diacylglycerol (DAG) production was important for proliferation in osteoblast-like MC3T3-E1 cells. The 1,2-DAG formation is mediated by various enzymes, such as phos-phoinositide (PI)-specific phospholipase C (PLC), phospholipase D (PLD), and phosphatidylcholine (PC)-specific phospholipase C (PC-PLC). In the present study, to elucidate the mechanism of the 1,2-DAG formation, we have examined the PGF2alpha-induced production of [(3)H]phosphorylcholine, a product of PC-PLC activity, in [(3)H]choline-labeled MC3T3-E1 cells. The PGF2alpha-induced [(3)H]phosphorylcholine production was inhibited by genistein, a potent protein tyrosine kinase inhibitor, and increased by vanadate, a potent protein tyrosine phosphatase inhibitor. However, there were no effects after treatment with protein kinase C (PKC) inhibitors, the guanosine triphosphate (GTP) binding protein activator, NaF/AlCl(3), a Ca(2+)-ionophore, or the potent activator of PKC, phorbol 12-myristate 13-acetate (PMA), suggesting that a tyrosine kinase(s) was involved in the PGF2alpha-induced [(3)H]phosphorylcholine formation. Furthermore, a PGF2alpha analogue, 16-(3-trifluoromethylphenoxy)-Omega-tetranor-trans-Delta(2) PGF2alpha methyl ester (ONO-995), stimulated the proliferation of MC3T3-E1 cells to a level similar to that seen with PGF2alpha, and also caused phosphorylcholine and 1,2-DAG generation. However, neither an increase in intracellular free calcium ion ([Ca(2+)]i) levels by PI-PLC, nor phosphatidylethanol formation (and choline production) by PC-PLD were observed. From these results, we conclude that PGF2alpha-induced 1,2-DAG accumulation was mediated mainly via tyrosine kinase(s)-dependent PC hydrolysis by PLC activity in osteoblast-like MC3T3-E1 cells. SN - 0914-8779 UR - https://www.unboundmedicine.com/medline/citation/15108061/Involvement_of_phosphatidylcholine_hydrolysis_by_phospholipase_C_in_prostaglandin_F2alpha_induced_12_diacylglycerol_formation_in_osteoblast_like_MC3T3_E1_cells_ DB - PRIME DP - Unbound Medicine ER -