title: Epigallocatechin-3-gallate elicits Ca2+ spike in MCF-7 breast cancer cells: essential role of Cav3.2 channels.
abstract: We used MCF-7 human breast cancer cells that endogenously express Cav3.1 and Cav3.2 T-type Ca(2+) channels toward a mechanistic study on the effect of EGCG on [Ca(2+)]i. Confocal Ca(2+) imaging showed that EGCG induces a [Ca(2+)]i spike which is due to extracellular Ca(2+) entry and is sensitive to catalase and to low-specificity (mibefradil) and high-specificity (Z944) T-type Ca(2+)channel blockers. siRNA knockdown of T-type Ca(2+) channels indicated the involvement of Cav3.2 but not Cav3.1. Application of EGCG to HEK cells expressing either Cav3.2 or Cav3.1 induced enhancement of Cav3.2 and inhibition of Cav3.1 channel activity. Measurements of K(+) currents in MCF-7 cells showed a reversible, catalase-sensitive inhibitory effect of EGCG, while siRNA for the Kv1.1 K(+) channel induced a reduction of the EGCG [Ca(2+)]i spike. siRNA for Cav3.2 reduced EGCG cytotoxicity to MCF-7 cells, as measured by calcein viability assay. Together, data suggest that EGCG promotes the activation of Cav3.2 channels through K(+) current inhibition leading to membrane depolarization, and in addition increases Cav3.2 currents. Cav3.2 channels are in part responsible for EGCG inhibition of MCF-7 viability, suggesting that deregulation of [Ca(2+)]i by EGCG may be relevant in breast cancer treatment.
authors.affiliation: Dipartimento di Scienze e Innovazione Tecnologica, DiSIT, Università del Piemonte Orientale, viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Elia
authors.initials: E
authors.lastname: Ranzato
authors.affiliation: Dipartimento di Scienze e Innovazione Tecnologica, DiSIT, Università del Piemonte Orientale, viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Magnelli
authors.affiliation: Dipartimento di Scienze e Innovazione Tecnologica, DiSIT, Università del Piemonte Orientale, viale T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Martinotti
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Rm 219 - 2185 East Mall, Vancouver, BC, Canada V6T 1Z4.
authors.forename: Zeina
authors.initials: Z
authors.lastname: Waheed
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Rm 219 - 2185 East Mall, Vancouver, BC, Canada V6T 1Z4.
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Cain
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Rm 219 - 2185 East Mall, Vancouver, BC, Canada V6T 1Z4.
authors.forename: Terrance P
authors.initials: TP
authors.lastname: Snutch
authors.affiliation: Istituto di Biofisica, Consiglio Nazionale delle Ricerche, via De Marini 6, 16149 Genova, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Marchetti
authors.affiliation: Dipartimento di Scienze e Innovazione Tecnologica, DiSIT, Università del Piemonte Orientale, viale T. Michel 11, 15121 Alessandria, Italy; Istituto di Biofisica, Consiglio Nazionale delle Ricerche, via De Marini 6, 16149 Genova, Italy. Electronic address: burlando@unipmn.it.
authors.forename: Bruno
authors.initials: B
authors.lastname: Burlando
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D020747:Calcium Channels, T-Type / Q000378:metabolism*; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D008564:Membrane Potentials / Q000187:drug effects*; D018408:Patch-Clamp Techniques
chemical_list: C479952:CACNA1H protein, human; D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D002392:Catechin; C045651:epigallocatechin gallate
keywords: Catalase; Cav3.1; Cav3.2; Confocal Ca(2+) imaging; Kv1.1; T-type Ca(2+) channels; Whole-cell voltage-clamp; Z944; [Ca(2+)](i) spike