title: Calcium dependence of purinergic subtype P2Y₁ receptor modulation of C6 glioma cell migration.
abstract: We have examined activation of purinergic P2Y₁ receptor-dependent Ca²⁺-signaling pathways in mediating C6 glioma cell migration. The administration of 2-methylthioadenosine 5'-diphosphate (2MeSADP), a selective agonist for P2Y₁R, induced marked increases in patterns of glioma migration in both scratch wound and Boyden chamber assays. Antagonism of P2Y₁R with either the broad spectrum purinergic blocker, pyridoxal-phosphate-6-azophenyl-2',4'-disulfonate (PPADS) or the specific P2Y₁R antagonist, 2'-deoxy-N⁶-methyladenosine-3',5'-bisphosphate (MRS2179), significantly inhibited C6 cell migration. Calcium-sensitive spectrofluorometry showed 2MeSADP stimulation of glioma cells caused a biphasic change in intracellular Ca²⁺ ([Ca²⁺]i). The rapid transient phase was unchanged in Ca²⁺-free solution reflecting a [Ca²⁺]i component due to intracellular stores release subsequent to activation of a metabotropic P2Y subtype receptor. The secondary prolonged phase of [Ca²⁺]i was abolished in Ca²⁺-free solution or in glioma cells treated with the store-operated channel (SOC) blocker, SKF96365. Treatment of glioma with either MRS2179 or PPADS significantly attenuated both the rapid and prolonged phases of [Ca²⁺]i. These results suggest critical roles for activation of P2Y₁R in mediating glioma cell mobility and migration with changes in [Ca²⁺]i contributing as a mechanistic link between activated receptor and functional response. Our findings suggest that pharmacological modulation of metabotropic P2Y₁R-dependent signaling pathways may serve as a novel therapeutic procedure to slow glioma progression.
authors.affiliation: Department of Anesthesiology, Pharmacology and Therapeutics, Faculty of Medicine, University of British Columbia, 2176 Health Sciences Mall, Vancouver, BC V6T1Z3 Canada.
authors.forename: Nattinee
authors.initials: N
authors.lastname: Jantaratnotai
authors.forename: James G
authors.initials: JG
authors.lastname: McLarnon
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D007424:Intracellular Fluid / Q000378:metabolism; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D009361:Neoplasm Invasiveness / Q000473:pathology; D058912:Purinergic P2Y Receptor Agonists / Q000494:pharmacology; D058921:Purinergic P2Y Receptor Antagonists / Q000494:pharmacology; D051381:Rats; D058472:Receptors, Purinergic P2Y1 / Q000187:drug effects / Q000502:physiology*; D013873:Thionucleotides / Q000494:pharmacology; D013997:Time Factors
chemical_list: D002121:Calcium Channel Blockers; D058912:Purinergic P2Y Receptor Agonists; D058921:Purinergic P2Y Receptor Antagonists; D058472:Receptors, Purinergic P2Y1; D013873:Thionucleotides; C002480:methylthio-ADP; D000244:Adenosine Diphosphate; D002118:Calcium