Nuclear localization of vasoactive intestinal peptide (VIP) receptors in human breast cancer

Vasoactive intestinal peptide (VIP) and its receptors (VPACs) are involved in proliferation, survival, and differentiation in human breast cancer cells. Its mechanism of action is traditionally thought to be through specific plasma membrane receptors. There is compelling evidence for a novel intracrine mode of genomic regulation by G-protein-coupled receptors (GPCRs) that implies both endocytosis and nuclear translocation of peripheral GPCR and/or the activation of nuclear-located GPCRs by endogenously-produced, non-secreted ligands. Regarding to VPAC receptors, which are GPCRs, there is only a report suggesting them as a dynamic system for signaling from plasma membrane and nuclear membrane complex. In this study, we show that VPAC(1) receptor is localized in cell nuclear fraction whereas VPAC(2) receptor presents an extranuclear localization and its protein expression is lower than that of VPAC(1) receptor in human breast tissue samples. Both receptors as well as VIP are overexpressed in breast cancer as compared to non-tumor tissue. Moreover, we report the markedly nuclear localization of VPAC(1) receptors in estrogen-dependent (T47D) and independent (MDA-MB-468) human breast cancer cell lines. VPAC(1) receptors are functional in plasma membrane and nucleus as shown by VIP stimulation of cAMP production in both cell lines. In addition, VIP increases its own intracellular and extracellular levels, and could be involved in the regulation of VPAC(1)-receptor traffic from the plasma membrane to the nucleus. These results support new concepts on function and regulation of nuclear GPCRs which could have an impact on development of new therapeutic drugs.

[1]  R. Jensen,et al.  VIP as a trophic factor in the CNS and cancer cells , 2003, Peptides.

[2]  A. Bajo,et al.  Transactivation of HER2 by vasoactive intestinal peptide in experimental prostate cancer: Antagonistic action of an analog of growth-hormone-releasing hormone. , 2007, International journal of oncology.

[3]  A. Couvineau,et al.  Molecular pharmacology and structure of VPAC Receptors for VIP and PACAP , 2002, Regulatory Peptides.

[4]  S. Choufani,et al.  Presence of functional endothelin-1 receptors in nuclear membranes of human aortic vascular smooth muscle cells. , 2000, Journal of cardiovascular pharmacology.

[5]  T. Hébert,et al.  Functional beta-adrenergic receptor signalling on nuclear membranes in adult rat and mouse ventricular cardiomyocytes. , 2006, Cardiovascular research.

[6]  M. Omary,et al.  Identification of nuclear receptors for VIP on a human colonic adenocarcinoma cell line. , 1987, Science.

[7]  T. Ho¨kfelt,et al.  Distribution and origin of peptide-containing nerve fibres in the rat and human mammary gland , 1996, Neuroscience.

[8]  R. Jensen,et al.  (N-stearyl, norleucine17) VIPhybrid is a broad spectrum vasoactive intestinal peptide receptor antagonist , 2002, Journal of Molecular Neuroscience.

[9]  A. Burkhalter,et al.  Activation of Metabotropic Glutamate Receptor mGlu5 on Nuclear Membranes Mediates Intranuclear Ca2+ Changes in Heterologous Cell Types and Neurons* , 2003, Journal of Biological Chemistry.

[10]  A. Bajo,et al.  Vasoactive intestinal peptide (VIP) induces transactivation of EGFR and HER2 in human breast cancer cells , 2009, Molecular and Cellular Endocrinology.

[11]  M. Cobb,et al.  ERK3 Is a Constitutively Nuclear Protein Kinase (*) , 1996, The Journal of Biological Chemistry.

[12]  A. Malviya,et al.  Mechanism regulating nuclear calcium signalingThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell. , 2006 .

[13]  E. Goetzl.  Diverse pathways for nuclear signaling by G protein-coupled receptors and their ligands. , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  M. Karin,et al.  Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. , 2000, Annual review of immunology.

[15]  A. Bajo,et al.  Vasoactive intestinal peptide behaves as a pro‐metastatic factor in human prostate cancer cells , 2009, The Prostate.

[16]  I. Gutiérrez-Cañas,et al.  Vasoactive intestinal peptide induces neuroendocrine differentiation in the LNCaP prostate cancer cell line through PKA, ERK, and PI3K , 2005, The Prostate.

[17]  A. Bajo,et al.  Vasoactive intestinal peptide (VIP) increases vascular endothelial growth factor (VEGF) expression and secretion in human breast cancer cells , 2007, Regulatory Peptides.

[18]  B. O'dowd,et al.  Agonist-independent Nuclear Localization of the Apelin, Angiotensin AT1, and Bradykinin B2 Receptors* , 2004, Journal of Biological Chemistry.

[19]  T. Moody,et al.  VIP-ellipticine derivatives inhibit the growth of breast cancer cells. , 2002, Life sciences.

[20]  E. Goetzl,et al.  Modulation of Pro-inflammatory Gene Expression by Nuclear Lysophosphatidic Acid Receptor Type-1* , 2003, Journal of Biological Chemistry.

[21]  T. Moody,et al.  Breast cancer growth is inhibited by vasoactive intestinal peptide (VIP) hybrid, a synthetic VIP receptor antagonist. , 1996, Cancer Research.

[22]  F. André,et al.  Chemokine receptor CXCR4 and early-stage non-small cell lung cancer: pattern of expression and correlation with outcome. , 2004, Annals of oncology : official journal of the European Society for Medical Oncology.

[23]  S. Choufani,et al.  G-protein-coupled receptors, channels, and Na+–H+ exchanger in nuclear membranes of heart, hepatic, vascular endothelial, and smooth muscle cellsThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell. , 2006 .

[24]  E. Goetzl.  Hypothesis: VPAC G protein-coupled receptors for vasoactive intestinal peptide constitute a dynamic system for signaling T cells from plasma membrane and nuclear membrane complexes , 2006, Regulatory Peptides.

[25]  K. Buchner.  Protein kinase C in the transduction of signals toward and within the cell nucleus. , 1995, European journal of biochemistry.

[26]  I. Gutiérrez-Cañas,et al.  Vasoactive intestinal peptide increases vascular endothelial growth factor expression and neuroendocrine differentiation in human prostate cancer LNCaP cells , 2004, Regulatory Peptides.

[27]  William L. Smith,et al.  Calcium-mediated Translocation of Cytosolic Phospholipase A2 to the Nuclear Envelope and Endoplasmic Reticulum (*) , 1995, The Journal of Biological Chemistry.

[28]  A. Gilman.  A protein binding assay for adenosine 3':5'-cyclic monophosphate. , 1970, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Crouch,et al.  Nuclear and cytoskeletal translocation and localization of heterotrimeric G‐proteins , 2000, Immunology and cell biology.

[30]  N. Heveker,et al.  G-protein-coupled receptors signalling at the cell nucleus: an emerging paradigm. , 2006, Canadian journal of physiology and pharmacology.

[31]  G. Bodega,et al.  Pituitary adenylate cyclase-activating peptide/vasoactive intestinal peptide receptors in human normal mammary gland and breast cancer tissue , 2005, Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology.

[32]  T. Moody,et al.  Vasoactive intestinal peptide receptors: a molecular target in breast and lung cancer. , 2007, Current pharmaceutical design.

[33]  I. Díaz-Laviada,et al.  Vasoactive intestinal peptide (VIP) induces c-fos expression in LNCaP prostate cancer cells through a mechanism that involves Ca2+ signalling. Implications in angiogenesis and neuroendocrine differentiation. , 2005, Biochimica et biophysica acta.

[34]  Z. Freyberg,et al.  Intracellular localization of phospholipase D1 in mammalian cells. , 2001, Molecular biology of the cell.

[35]  D. Jackson,et al.  Coupled Transcription and Translation Within Nuclei of Mammalian Cells , 2001, Science.

[36]  K. Kawamura,et al.  Intracellular distribution of adenylate cyclase in human cardiocytes determined by electron microscopic cytochemistry , 1998, Microscopy research and technique.

[37]  A. Hoffmann,et al.  Nuclear Localization of IκBα Is Mediated by the Second Ankyrin Repeat: the IκBα Ankyrin Repeats Define a Novel Class of cis-Acting Nuclear Import Sequences , 1998, Molecular and Cellular Biology.

[38]  Susan S. Taylor,et al.  A-kinase-interacting protein localizes protein kinase A in the nucleus. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[39]  A. Couvineau,et al.  Class II G protein-coupled receptors for VIP and PACAP: Structure, models of activation and pharmacology , 2007, Peptides.

[40]  M. Leite,et al.  Calcium signaling in the nucleus. , 2006, Canadian journal of physiology and pharmacology.

[41]  M. Madsen,et al.  Estradiol down regulates expression of vasoactive intestinal polypeptide receptor type-1 in breast cancer cell lines , 2001, Molecular and Cellular Endocrinology.

[42]  K. Riesbeck,et al.  A novel localization of the G-protein-coupled CysLT1 receptor in the nucleus of colorectal adenocarcinoma cells. , 2005, Cancer research.

[43]  J C Reubi,et al.  Neuropeptide receptors in health and disease: the molecular basis for in vivo imaging. , 1995, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[44]  E. Wickstrom,et al.  Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase activating peptide (PACAP) receptor specific peptide analogues for PET imaging of breast cancer: In vitro/in vivo evaluation , 2007, Regulatory Peptides.

[45]  Fude Fang,et al.  Two domains are critical for the nuclear localization of soluble adenylyl cyclase. , 2006, Biochimie.

[46]  J. Waschek,et al.  Differential expression of VIP/PACAP receptor genes in breast, intestinal, and pancreatic cell lines. , 1995, Cancer letters.

[47]  D Rodbard,et al.  Ligand: a versatile computerized approach for characterization of ligand-binding systems. , 1980, Analytical biochemistry.

[48]  A. Ribeiro-da-Silva,et al.  Nuclear prostaglandin signaling system: biogenesis and actions via heptahelical receptors. , 2003, Canadian journal of physiology and pharmacology.

[49]  S. Choufani,et al.  Proinflammatory Gene Induction by Platelet-Activating Factor Mediated Via Its Cognate Nuclear Receptor1 , 2002, The Journal of Immunology.

[50]  S. Rhee,et al.  The Role of Carboxyl-terminal Basic Amino Acids in Gqα-dependent Activation, Particulate Association, and Nuclear Localization of Phospholipase C-β1* , 1996, The Journal of Biological Chemistry.

[51]  S. Molotchnikoff,et al.  Regulation of eNOS Expression in Brain Endothelial Cells by Perinuclear EP3 Receptors , 2002, Circulation research.

[52]  R. Jensen,et al.  Breast Cancer VPAC1 Receptors , 2006, Annals of the New York Academy of Sciences.

[53]  A. Martelli,et al.  The nuclear phosphoinositide 3-kinase/AKT pathway: a new second messenger system. , 2002, Biochimica et biophysica acta.

[54]  N. Heveker,et al.  Nitric Oxide Signaling via Nuclearized Endothelial Nitric-oxide Synthase Modulates Expression of the Immediate Early Genes iNOS and mPGES-1* , 2006, Journal of Biological Chemistry.

[55]  Hung-Ming Wang,et al.  A reverse transcription comparative real-time PCR method for quantitative detection of angiogenic growth factors in head and neck cancer patients. , 2002, Clinical biochemistry.