Sustained activation of Erk1/2 MAPK and cell growth suppression by the insert-negative, but not the insert-positive isoform of the human calcitonin receptor.

Recently we reported that calcitonin (CT) induces growth arrest at the G2 stage of the cell cycle in HEK-293 cell lines expressing the most abundant, insert-negative, isoform of the human CT receptor (insert -ve hCTR). The present study investigates the involvement of the MAPK signalling pathway in the anti-proliferative actions of CT and compares the activity of an isoform of the hCTR that contains a 16 amino acid insert in the first putative intracellular loop (insert +ve hCTR). Comparison of HEK-293 cells stably transfected with the insert -ve or the insert +ve hCTR, showed that accumulation of cAMP and intracellular free calcium in response to CT were specific for the insert -ve receptor isoform. However, a novel acidification of the extracellular medium was mediated by both isoforms. Treatment with CT of cells expressing the insert -ve hCTR, caused a decrease in cell growth associated with an induction of p21(WAF1/CIP1). Analysis by fluorescence-activated cell scanning showed that growth inhibition was associated with an accumulation of cells in G2. CT treatment of cells expressing the insert -ve, but not insert +ve hCTR, induced the phosphorylation of Erk1/2 MAPK, which persisted for at least 72 h. Treatment of cells expressing the insert -ve hCTR with the MAPK kinase (MEK) inhibitor, PD-98059, inhibited the phosphorylation of Erk1/2 and abrogated the growth inhibitory effects of salmon CT, the accumulation of cells in G2, and the associated induction of p21(WAF1/CIP1). These data suggest that activation of Erk1/2 are downstream effectors of the insert -ve hCTR in modulating cell cycle progression.

[1]  P Hiselius,et al.  The New Generation , 2021, The Women's Liberation Movement in Russia.

[2]  A. Evdokiou,et al.  Calcitonin receptor-mediated growth suppression of HEK-293 cells is accompanied by induction of p21WAF1/CIP1 and G2/M arrest. , 1999, Molecular endocrinology.

[3]  R. Baron,et al.  Cytoskeleton-dependent tyrosine phosphorylation of the p130(Cas) family member HEF1 downstream of the G protein-coupled calcitonin receptor. Calcitonin induces the association of HEF1, paxillin, and focal adhesion kinase. , 1999, The Journal of biological chemistry.

[4]  R. Baron,et al.  Cytoskeleton-dependent Tyrosine Phosphorylation of the p130Cas Family Member HEF1 Downstream of the G Protein-coupled Calcitonin Receptor , 1999, The Journal of Biological Chemistry.

[5]  M. Matsuda,et al.  Activation of the ERK/MAPK pathway by an isoform of rap1GAP associated with Gαi , 1999, Nature.

[6]  Lakshmi A. Devi,et al.  G-protein-coupled receptor heterodimerization modulates receptor function , 1999, Nature.

[7]  R. Seger,et al.  Gi-mediated activation of mitogen-activated protein kinase (MAPK) pathway by receptor mimetic basic secretagogues of connective tissue-type mast cells: bifurcation of arachidonic acid-induced release upstream of MAPK. , 1999, The Journal of pharmacology and experimental therapeutics.

[8]  R. Baron,et al.  Protein kinase C antagonizes pertussis-toxin-sensitive coupling of the calcitonin receptor to adenylyl cyclase. , 1999, European journal of biochemistry.

[9]  P. Fisher,et al.  α‐Adrenergic inhibition of proliferation in HepG2 cells stably transfected with the α1B‐adrenergic receptor through a p42MAP kinase/p21Cip1/WAF1‐dependent pathway , 1998 .

[10]  J. Gutkind,et al.  Cell growth control by G protein-coupled receptors: from signal transduction to signal integration , 1998, Oncogene.

[11]  R. Baron,et al.  The Calcitonin Receptor Stimulates Shc Tyrosine Phosphorylation and Erk1/2 Activation , 1998, The Journal of Biological Chemistry.

[12]  D. Galson,et al.  Phospholipase D- and protein kinase C isoenzyme-dependent signal transduction pathways activated by the calcitonin receptor. , 1998, Endocrinology.

[13]  P. Dent,et al.  The mitogen-activated protein (MAP) kinase cascade can either stimulate or inhibit DNA synthesis in primary cultures of rat hepatocytes depending upon whether its activation is acute/phasic or chronic. , 1998, The Biochemical journal.

[14]  P. Sexton,et al.  Purification of calcitonin-like peptides from rat brain and pituitary. , 1998, Endocrinology.

[15]  T. Martin,et al.  Calcitonin receptors, bone sialoprotein and osteopontin are expressed in primary breast cancers , 1997, International journal of cancer.

[16]  G. Belinsky,et al.  A New Action of Parathyroid Hormone , 1997, The Journal of Biological Chemistry.

[17]  A Sewing,et al.  High-intensity Raf signal causes cell cycle arrest mediated by p21Cip1 , 1997, Molecular and cellular biology.

[18]  E. Lees,et al.  Raf-induced proliferation or cell cycle arrest is determined by the level of Raf activity with arrest mediated by p21Cip1 , 1997, Molecular and cellular biology.

[19]  M. Lacroix,et al.  Regulation of c-fos and c-jun Expression by Calcitonin in Human Breast Cancer Cells , 1997, Calcified Tissue International.

[20]  D. Tindall,et al.  Effects of the calciotrophic peptides calcitonin and parathyroid hormone on prostate cancer growth and chemotaxis , 1997, The Prostate.

[21]  K. Pumiglia,et al.  Cell cycle arrest mediated by the MEK/mitogen-activated protein kinase pathway. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[22]  W. Samson,et al.  Calcitonin is a physiological inhibitor of prolactin secretion in ovariectomized female rats. , 1996, Endocrinology.

[23]  K. Beaumont,et al.  Molecular cloning and functional expression of a third isoform of the human calcitonin receptor and partial characterization of the calcitonin receptor gene. , 1995, Endocrinology.

[24]  P. Sexton,et al.  Functionally different isoforms of the human calcitonin receptor result from alternative splicing of the gene transcript. , 1995, Molecular endocrinology.

[25]  S. Krane,et al.  Expression of two human skeletal calcitonin receptor isoforms cloned from a giant cell tumor of bone. The first intracellular domain modulates ligand binding and signal transduction. , 1995, The Journal of clinical investigation.

[26]  M. Gershengorn,et al.  Inhibition of inositol phosphate second messenger formation by intracellular loop one of a human calcitonin receptor. Expression and mutational analysis of synthetic receptor genes. , 1994, The Journal of biological chemistry.

[27]  N. Ahn,et al.  Transformation of mammalian cells by constitutively active MAP kinase kinase. , 1994, Science.

[28]  D. Thompson,et al.  Cloning and characterization of an abundant subtype of the human calcitonin receptor. , 1994, Molecular pharmacology.

[29]  P. Cohen,et al.  EGF triggers neuronal differentiation of PC12 cells that overexpress the EGF receptor , 1994, Current Biology.

[30]  P. Sexton,et al.  Isoforms of the rat calcitonin receptor: consequences for ligand binding and signal transduction. , 1994, Endocrinology.

[31]  C. Marshall,et al.  Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells , 1994, Cell.

[32]  W. Rayford,et al.  Calcitonin stimulates growth of human prostate cancer cells through receptor-mediated increase in cyclic adenosine 3',5'-monophosphates and cytoplasmic Ca2+ transients. , 1994, Endocrinology.

[33]  D. Thompson,et al.  A recombinant human calcitonin receptor functions as an extracellular calcium sensor. , 1993, The Journal of biological chemistry.

[34]  W. H. Betts,et al.  The substance P fragment SP-(7-11) increases prostaglandin E2, intracellular Ca2+ and collagenase production in bovine articular chondrocytes. , 1993, The Biochemical journal.

[35]  H. Lodish,et al.  Cloning, characterization, and expression of a human calcitonin receptor from an ovarian carcinoma cell line. , 1992, The Journal of clinical investigation.

[36]  Y. Cho‐Chung.  Role of cyclic AMP receptor proteins in growth, differentiation, and suppression of malignancy: new approaches to therapy. , 1990, Cancer research.

[37]  T. Martin,et al.  Human placental calcitonin receptors. , 1988, The Biochemical journal.

[38]  R. Tsien,et al.  A new generation of Ca2+ indicators with greatly improved fluorescence properties. , 1985, The Journal of biological chemistry.

[39]  T. Martin,et al.  Characteristics of selective activation of cyclic AMP-dependent protein kinase isoenzymes by calcitonin and prostaglandin E2 in human breast cancer cells. , 1984, The Biochemical journal.

[40]  T. Martin,et al.  Calcitonin effects on growth and on selective activation of type II isoenzyme of cyclic adenosine 3':5'-monophosphate-dependent protein kinase in T 47D human breast cancer cells. , 1983, Cancer research.

[41]  T. Chambers,et al.  Calcitonin alters behaviour of isolated osteoclasts , 1982, The Journal of pathology.

[42]  S. Sagar,et al.  Calcitonin: regional distribution of the hormone and its binding sites in the human brain and pituitary. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[43]  T. Martin,et al.  Calcitonin binding and degradation by two cultured human breast cancer cell lines (MCF 7 and T 47D). , 1981, The Biochemical journal.

[44]  J. Eisman,et al.  Calcitonin and 1,25-dihydroxyvitamin D3 receptors in human breast cancer cell lines. , 1980, Cancer research.

[45]  T. Martin,et al.  Properties of a calcitonin receptor and adenylate cyclase in BEN cells, a human cancer cell line. , 1980, Cancer research.

[46]  Richard Axel,et al.  Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells , 1977, Cell.

[47]  S. Marx,et al.  Calcitonin Receptors of Kidney and Bone , 1972, Science.

[48]  M. Pondel,et al.  The porcine calcitonin receptor promoter directs expression of a linked reporter gene in a tissue and developmental specific manner in transgenic mice. , 1999, Endocrinology.

[49]  I. Bagchi,et al.  Attenuation of calcitonin gene expression in pregnant rat uterus leads to a block in embryonic implantation. , 1998, Endocrinology.

[50]  P. Dent,et al.  Alpha-adrenergic inhibition of proliferation in HepG2 cells stably transfected with the alpha1B-adrenergic receptor through a p42MAPkinase/p21Cip1/WAF1-dependent pathway. , 1998, FEBS letters.

[51]  C. Blake.  Hormone receptors. , 1978, Endeavour.