Abstract
In the present study, we have assessed whether a putative calcium channel α2δ2 auxiliary subunit (CACNA2D2 gene) could be involved in prostate cancer (PCA) progression. We therefore carried out experiments to determine whether this protein is expressed in PCA LNCaP cells and in PCA tissues, and whether its expression may be altered during cancer development. In addition, we evaluated the influence on cell proliferation of overexpressing or downregulating this subunit. In vitro experiments show that α2δ2 subunit overexpression is associated with increased cell proliferation, alterations of calcium homeostasis and the recruitment of a nuclear factor of activated T-cells pathway. Furthermore, we carried out in vivo experiments on immuno-deficient nude mice in order to evaluate the tumorigenic potency of the α2δ2 subunit. We show that α2δ2-overexpressing PCA LNCaP cells are more tumorigenic than control LNCaP cells when injected into nude mice. In addition, gabapentin, a ligand of α2δ2, reduces tumor development in LNCaP xenografts. Finally, we show that the action of α2δ2 on tumor development occurs not only through a stimulation of proliferation, but also through a stimulation of angiogenesis, via an increased secretion of vascular endothelial growth factor in cells overexpressing α2δ2.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Bidaud I, Mezghrani A, Swayne LA, Monteil A, Lory P . Voltage-gated calcium channels in genetic diseases. Biochim Biophys Acta 2006; 1763: 1169–1174.
Monteith GR, Davis FM, Roberts-Thomson SJ . Calcium channels and pumps in cancer: changes and consequences. J Biol Chem 2012; 287: 31666–31673.
Lee JM, Davis FM, Roberts-Thomson SJ, Monteith GR . Ion channels and transporters in cancer. 4. Remodeling of Ca(2+) signaling in tumorigenesis: role of Ca(2+) transport. Am J Physiol Cell Physiol 2011; 301: C969–C976.
Wang XT, Nagaba Y, Cross HS, Wrba F, Zhang L, Guggino SE . The mRNA of L-type calcium channel elevated in colon cancer: protein distribution in normal and cancerous colon. Am J Pathol 2000; 157: 1549–1562.
Toyota M, Ho C, Ohe-Toyota M, Baylin SB, Issa JP . Inactivation of CACNA1G, a T-type calcium channel gene, by aberrant methylation of its 5' CpG island in human tumors. Cancer Res 1999; 59: 4535–4541.
Fiske JL, Fomin VP, Brown ML, Duncan RL, Sikes RA . Voltage-sensitive ion channels and cancer. Cancer Metastasis Rev 2006; 25: 493–500.
Kunzelmann K . Ion channels and cancer. J Membr Biol 2005; 205: 159–173.
Bennett ES, Smith BA, Harper JM . Voltage-gated Na+ channels confer invasive properties on human prostate cancer cells. Pflugers Arch 2004; 447: 908–914.
Mariot P, Vanoverberghe K, Lalevee N, Rossier MF, Prevarskaya N . Overexpression of an alpha 1H (Cav3.2) T-type calcium channel during neuroendocrine differentiation of human prostate cancer cells. J Biol Chem 2002; 277: 10824–10833.
Skryma R, Van Coppenolle F, Dufy-Barbe L, Dufy B, Prevarskaya N . Characterization of Ca(2+)-inhibited potassium channels in the LNCaP human prostate cancer cell line. Receptors Channels 1999; 6: 241–253.
Spitzner M, Ousingsawat J, Scheidt K, Kunzelmann K, Schreiber R . Voltage-gated K+ channels support proliferation of colonic carcinoma cells. Faseb J 2007; 21: 35–44.
Gackiere F, Bidaux G, Delcourt P, Van Coppenolle F, Katsogiannou M, Dewailly E et al. CaV3.2 T-type calcium channels are involved in calcium-dependent secretion of neuroendocrine prostate cancer cells. J Biol Chem 2008; 283: 10162–10173.
Lerman MI, Minna JD . The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3: identification and evaluation of the resident candidate tumor suppressor genes. The International Lung Cancer Chromosome 3p21.3 Tumor Suppressor Gene Consortium. Cancer Res 2000; 60: 6116–6133.
Ghosh S, Ghosh A, Maiti GP, Alam N, Roy A, Roy B et al. Alterations of 3p21.31 tumor suppressor genes in head and neck squamous cell carcinoma: Correlation with progression and prognosis. Int J Cancer 2008; 123: 2594–2604.
Wanajo A, Sasaki A, Nagasaki H, Shimada S, Otsubo T, Owaki S et al. Methylation of the calcium channel-related gene, CACNA2D3, is frequent and a poor prognostic factor in gastric cancer. Gastroenterology 2008; 135: 580–590.
Mitra R, Lee J, Jo J, Milani M, McClintick JN, Edenberg HJ et al. Prediction of postoperative recurrence-free survival in non-small cell lung cancer by using an internationally validated gene expression model. Clin Cancer Res 2011; 17: 2934–2946.
Halatsch ME, Low S, Mursch K, Hielscher T, Schmidt U, Unterberg A et al. Candidate genes for sensitivity and resistance of human glioblastoma multiforme cell lines to erlotinib. Laboratory investigation. J Neurosurg 2009; 111: 211–218.
Carboni GL, Gao B, Nishizaki M, Xu K, Minna JD, Roth JA et al. CACNA2D2-mediated apoptosis in NSCLC cells is associated with alterations of the intracellular calcium signaling and disruption of mitochondria membrane integrity. Oncogene 2003; 22: 615–626.
Dooley DJ, Taylor CP, Donevan S, Feltner D . Ca2+ channel alpha2delta ligands: novel modulators of neurotransmission. Trends Pharmacol Sci 2007; 28: 75–82.
Li Z, Taylor CP, Weber M, Piechan J, Prior F, Bian F et al. Pregabalin is a potent and selective ligand for alpha(2)delta-1 and alpha(2)delta-2 calcium channel subunits. Eur J Pharmacol 2011; 667: 80–90.
Starborg M, Gell K, Brundell E, Hoog C . The murine Ki-67 cell proliferation antigen accumulates in the nucleolar and heterochromatic regions of interphase cells and at the periphery of the mitotic chromosomes in a process essential for cell cycle progression. J Cell Sci 1996; 109: 143–153.
Dolphin AC . Calcium channel auxiliary alpha2delta and beta subunits: trafficking and one step beyond. Nat Rev Neurosci 2012; 13: 542–555.
Liu C, Hermann TE . Characterization of ionomycin as a calcium ionophore. J Biol Chem 1978; 253: 5892–5894.
Lytton J, Westlin M, Hanley MR . Thapsigargin inhibits the sarcoplasmic or endoplasmic reticulum Ca-ATPase family of calcium pumps. J Biol Chem 1991; 266: 17067–17071.
Davies A, Hendrich J, Van Minh AT, Wratten J, Douglas L, Dolphin AC . Functional biology of the alpha(2)delta subunits of voltage-gated calcium channels. Trends Pharmacol Sci 2007; 28: 220–228.
da Costa Prando E, Cavalli LR, Rainho CA . Evidence of epigenetic regulation of the tumor suppressor gene cluster flanking RASSF1 in breast cancer cell lines. Epigenetics 2011; 6: 1413–1424.
Zhao W, Wang L, Han H, Jin K, Lin N, Guo T et al. 1B50-1, a mAb raised against recurrent tumor cells, targets liver tumor-initiating cells by binding to the calcium channel alpha2delta1 subunit. Cancer Cell 2013; 23: 541–556.
Negrini S, Prada I, D'Alessandro R, Meldolesi J . REST: an oncogene or a tumor suppressor? Trends Cell Biol 2013; 23: 289–295.
Pelosi G, Fumagalli C, Trubia M, Sonzogni A, Rekhtman N, Maisonneuve P et al. Dual role of RASSF1 as a tumor suppressor and an oncogene in neuroendocrine tumors of the lung. Anticancer Res 2010; 30: 4269–4281.
Hobom M, Dai S, Marais E, Lacinova L, Hofmann F, Klugbauer N . Neuronal distribution and functional characterization of the calcium channel alpha2delta-2 subunit. Eur J Neurosci 2000; 12: 1217–1226.
Dolphin AC, Wyatt CN, Richards J, Beattie RE, Craig P, Lee JH et al. The effect of alpha2-delta and other accessory subunits on expression and properties of the calcium channel alpha1G. J Physiol 1999; 519: 35–45.
Gackiere F, Warnier M, Katsogiannou M, Derouiche S, Delcourt P, Dewailly E et al. Functional coupling between large-conductance potassium channels and Cav3.2 voltage-dependent calcium channels participates in prostate cancer cell growth. Biol Open 2013; 2: 941–951.
Canti C, Nieto-Rostro M, Foucault I, Heblich F, Wratten J, Richards MW et al. The metal-ion-dependent adhesion site in the Von Willebrand factor-A domain of alpha2delta subunits is key to trafficking voltage-gated Ca2+ channels. Proc Natl Acad Sci USA 2005; 102: 11230–11235.
Whittaker CA, Hynes RO . Distribution and evolution of von Willebrand/integrin A domains: widely dispersed domains with roles in cell adhesion and elsewhere. Mol Biol Cell 2002; 13: 3369–3387.
Eroglu C, Allen NJ, Susman MW, O'Rourke NA, Park CY, Ozkan E et al. Gabapentin receptor alpha2delta-1 is a neuronal thrombospondin receptor responsible for excitatory CNS synaptogenesis. Cell 2009; 139: 380–392.
Garcia K, Nabhani T, Garcia J . The calcium channel alpha2/delta1 subunit is involved in extracellular signalling. J Physiol 2008; 586: 727–738.
Bornstein P . Thrombospondins function as regulators of angiogenesis. J Cell Commun Signal 2009; 3: 189–200.
Kondratska K, Kondratskyi A, Yassine M, Lemonnier L, Lepage G, Morabito A et al. Orai1 and STIM1 mediate SOCE and contribute to apoptotic resistance of pancreatic adenocarcinoma. Biochim Biophys Acta 2014; 1843: 2263–2269.
Chomczynski P, Sacchi N . Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156–159.
Katsogiannou M, El Boustany C, Gackiere F, Delcourt P, Athias A, Mariot P et al. Caveolae contribute to the apoptosis resistance induced by the alpha(1A)-adrenoceptor in androgen-independent prostate cancer cells. PLoS One 2009; 4: e7068.
Gackiere F, Bidaux G, Lory P, Prevarskaya N, Mariot P . A role for voltage gated T-type calcium channels in mediating "capacitative" calcium entry? Cell Calcium 2006; 39: 357–366.
Oh C, Park S, Lee EK, Yoo YJ . Downregulation of ubiquitin level via knockdown of polyubiquitin gene Ubb as potential cancer therapeutic intervention. Sci Rep 2013; 3: 2623.
Wilhelm MT, Rufini A, Wetzel MK, Tsuchihara K, Inoue S, Tomasini R et al. Isoform-specific p73 knockout mice reveal a novel role for delta Np73 in the DNA damage response pathway. Genes Dev 2010; 24: 549–560.
Lim DJ, Liu XL, Sutkowski DM, Braun EJ, Lee C, Kozlowski JM . Growth of an androgen-sensitive human prostate cancer cell line, LNCaP, in nude mice. Prostate 1993; 22: 109–118.
Zour E, Lodhi SA, Nesbitt RU, Silbering SB, Chaturvedi PR . Stability studies of gabapentin in aqueous solutions. Pharm Res 1992; 9: 595–600.
Acknowledgements
We thank the members of the imaging platform BICEL, E Richard and C Slommiany for their helpful contribution. This work was supported by INSERM, the University of Lille1 and the Region Nord-Pas de Calais. This work was supported by INSERM, the University of Lille1 and the Region Nord-Pas de Calais.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies this paper on the Oncogene website
Rights and permissions
About this article
Cite this article
Warnier, M., Roudbaraki, M., Derouiche, S. et al. CACNA2D2 promotes tumorigenesis by stimulating cell proliferation and angiogenesis. Oncogene 34, 5383–5394 (2015). https://doi.org/10.1038/onc.2014.467
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2014.467
This article is cited by
-
Identifying tumour microenvironment-related signature that correlates with prognosis and immunotherapy response in breast cancer
Scientific Data (2023)
-
The Role οf Ion Channels in the Development and Progression of Prostate Cancer
Molecular Diagnosis & Therapy (2023)
-
Calcium signaling in neurodevelopment and pathophysiology of autism spectrum disorders
Molecular Biology Reports (2022)
-
Calcium channelopathies and intellectual disability: a systematic review
Orphanet Journal of Rare Diseases (2021)
-
Transcriptional and epigenetic landscape of Ca2+-signaling genes in hepatocellular carcinoma
Journal of Cell Communication and Signaling (2021)