Molecular Human Reproduction, Vol. 6, No. 5, 435-442,
May 2000
© 2000 European Society of Human Reproduction and Embryology
Ovary and oogenesis |
Characterization of calcium-mobilizing, purinergic P2Y2 receptors in human ovarian cancer cells
1 Department of Obstetrics and Gynecology, University of Lubeck, Germany, D-23538 and 2 Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
Abstract
In human ovarian EFO-21 and EFO-27 carcinoma cells, extracellular ATP induced a concentration-dependent rise in intracellular calcium concentration ([Ca2+]i), suggesting the expression of a purinoreceptor. ATP and UTP were equipotent in generating [Ca2+]i signals, followed by ATP-
-S and ADP, whereas ß,-ATP, 2 methyl 1 thio-ATP, 3'-o-(4-benzoyl) benzoyl-ATP, AMP, and adenosine were ineffective. This pharmacological profile suggested the presence of the P2Y2 subtype in both cell types, and this was confirmed by reverse transcription–polymerase chain reaction (RT–PCR) analysis using P2Y2 primers. ATP-induced [Ca2+]i signals were composed of two phases: an early and extracellular calcium-independent phase, followed by a sustained plateau phase that was dependent on capacitative calcium influx. In addition to the rise in the [Ca2+]i, a time- and concentration-dependent increase in phosphatidylethanol accumulation was observed in ATP-stimulated cells, indicating an increase in phospholipase D activity. RT–PCR analysis identified the expression of a transcript for the phospholipase D-1 subtype of this enzyme. Activation of these receptors by a slowly degradable analogue, ATP--S, attenuated basal and fetal calf serum-induced cell proliferation in a time- and concentration-dependent manner. These results indicate that ATP may act as an extracellular messenger in controlling the ovarian epithelial cell cycle through P2Y2 receptors.
ATP receptors/calcium/cell proliferation/phospholipase C
Notes
3 To whom correspondence should be addressed at: Section on Cellular Signaling, ERRB/NICHD, Bldg. 49, Room 6A-36, 49 Convent Drive, Bethesda, MD 20892-4510, USA
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