Characterization of calcium-mobilizing, purinergic P2Y(2) receptors in human ovarian cancer cells.
In human ovarian EFO-21 and EFO-27 carcinoma cells, extracellular ATP induced a concentration-dependent rise in intracellular calcium concentration ([Ca(2+)](i)), suggesting the expression of a purinoreceptor. ATP and UTP were equipotent in generating [Ca(2+)](i) signals, followed by ATP-gamma-S and ADP, whereas beta, gamma-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 P2Y(2) subtype in both cell types, and this was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) analysis using P2Y(2) primers. ATP-induced [Ca(2+)](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 [Ca(2+)](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-gamma-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 P2Y(2) receptors.
Schultze-Mosgau
A
A
Department of Obstetrics and Gynecology, University of Lubeck, Germany.
Katzur
A C
AC
Arora
K K
KK
Stojilkovic
S S
SS
Diedrich
K
K
Ortmann
O
O
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D004095:Dihydropyridines / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D020404:Glycerophospholipids / Q000378:metabolism; D006801:Humans; D009543:Nifedipine / Q000494:pharmacology; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D010739:Phospholipase D / Q000187:drug effects / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D058473:Receptors, Purinergic P2Y2; D019284:Thapsigargin / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014544:Uridine Triphosphate / Q000494:pharmacology
D002121:Calcium Channel Blockers; D004095:Dihydropyridines; D004791:Enzyme Inhibitors; D020404:Glycerophospholipids; C546385:P2RY2 protein, human; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; C051521:phosphatidylethanol; C022571:adenosine 5'-O-(3-thiotriphosphate); D019284:Thapsigargin; C038806:1,4-dihydropyridine; D000255:Adenosine Triphosphate; D010739:Phospholipase D; D009543:Nifedipine; D011188:Potassium; D002118:Calcium; D014544:Uridine Triphosphate