title: Store-operated Ca2+ entry does not control proliferation in primary cultures of human metastatic renal cellular carcinoma.
abstract: Store-operated Ca(2+) entry (SOCE) is activated following depletion of the inositol-1,4,5-trisphosphate (InsP3)-sensitive Ca(2+) pool to regulate proliferation in immortalized cell lines established from either primary or metastatic lesions. The molecular nature of SOCE may involve both Stim1, which senses Ca(2+) levels within the endoplasmic reticulum (ER) Ca(2+) reservoir, and a number of a Ca(2+)-permeable channels on the plasma membrane, including Orai1, Orai3, and members of the canonical transient receptor (TRPC1-7) family of ion channels. The present study was undertaken to assess whether SOCE is expressed and controls proliferation in primary cultures isolated from secondary lesions of heavily pretreated metastatic renal cell carcinoma (mRCC) patients. SOCE was induced following pharmacological depletion of the ER Ca(2+) store, but not by InsP3-dependent Ca(2+) release. Metastatic RCC cells express Stim1-2, Orai1-3, and TRPC1-7 transcripts and proteins. In these cells, SOCE was insensitive to BTP-2, 10 µM Gd(3+) and Pyr6, while it was inhibited by 100 µM Gd(3+), 2-APB, and carboxyamidotriazole (CAI). Neither Gd(3+) nor 2-APB or CAI impaired mRCC cell proliferation. Consistently, no detectable Ca(2+) signal was elicited by growth factor stimulation. Therefore, a functional SOCE is expressed but does not control proliferation of mRCC cells isolated from patients resistant to multikinase inhibitors.
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Silvia
authors.identifier: 0000-0002-9107-6291
authors.initials: S
authors.lastname: Dragoni
authors.affiliation: Laboratory of Immunology Transplantation, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Turin
authors.affiliation: Department of Molecular Medicine, University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Umberto
authors.identifier: 0000-0003-2951-5983
authors.initials: U
authors.lastname: Laforenza
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Duilio Michele
authors.initials: DM
authors.lastname: Potenza
authors.affiliation: Laboratory of Immunology Transplantation, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Bottino
authors.affiliation: Christian Doppler Laboratory for Flow Chemistry, Institute of Chemistry, Karl-Franzens-University Graz, Heinrichstrasse 28, 8010 Graz, Austria.
authors.forename: Toma N
authors.initials: TN
authors.lastname: Glasnov
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Martina
authors.identifier: 0000-0001-8019-0497
authors.initials: M
authors.lastname: Prestia
authors.affiliation: Laboratory of Immunology Transplantation, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ferulli
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Anna
authors.identifier: 0000-0001-8805-5126
authors.initials: A
authors.lastname: Saitta
authors.affiliation: Medical Oncology, "Maggiore della Carità" University Hospital, 28100 Novara, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Mosca
authors.affiliation: Department of Health Sciences, University of Molise, Via F. de Santis, 86100 Campobasso, Italy.
authors.forename: Germano
authors.initials: G
authors.lastname: Guerra
authors.affiliation: Laboratory of Biotechnology, Center for the Study of Myelofibrosis, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Rosti
authors.affiliation: Department of Pathological Anatomy, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Ombretta
authors.initials: O
authors.lastname: Luinetti
authors.affiliation: Medical Oncology, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Carlo
authors.identifier: 0000-0002-5839-3965
authors.initials: C
authors.lastname: Ganini
authors.affiliation: Medical Oncology, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Camillo
authors.initials: C
authors.lastname: Porta
authors.affiliation: Medical Oncology, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Pedrazzoli
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Franco
authors.identifier: 0000-0001-6998-1095
authors.initials: F
authors.lastname: Tanzi
authors.affiliation: Laboratory of Immunology Transplantation, Foundation IRCCS Policlinico San Matteo, Piazzale Golgi 19, 27100 Pavia, Italy ; Department of Clinical-Surgical, Diagnostic and Pediatric Sciences, University of Pavia, 27100 Pavia, Italy.
authors.forename: Daniela
authors.identifier: 0000-0002-3182-266X
authors.initials: D
authors.lastname: Montagna
authors.affiliation: Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia, Via Forlanini 6, 27100 Pavia, Italy ; Department of Physiology, University of Pavia, Via Forlanini 6, 27100 Pavia, Italy.
authors.forename: Francesco
authors.identifier: 0000-0003-0010-0098
authors.initials: F
authors.lastname: Moccia
mesh_terms: D000368:Aged; D015220:Calcium Channels / Q000096:biosynthesis; D020013:Calcium Signaling / Q000235:genetics*; D002292:Carcinoma, Renal Cell / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D049109:Cell Proliferation / Q000235:genetics*; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000473:pathology; D005260:Female; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis; D008875:Middle Aged; D009362:Neoplasm Metastasis / Q000235:genetics* / Q000473:pathology; D009363:Neoplasm Proteins / Q000096:biosynthesis; D000071740:ORAI1 Protein; D061251:Primary Cell Culture; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D000071737:Stromal Interaction Molecule 1; D050052:TRPC Cation Channels
chemical_list: D015220:Calcium Channels; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D000071740:ORAI1 Protein; C510746:ORAI1 protein, human; C522867:Orai3 protein, human; D047428:Protein Kinase Inhibitors; C103530:STIM1 protein, human; D000071737:Stromal Interaction Molecule 1; D050052:TRPC Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1; D015544:Inositol 1,4,5-Trisphosphate