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All rights reserved. http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/dc/elements/1.1/title BEL Framework Large Corpus Document http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/pav/authoredBy http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#_5 http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://purl.org/pav/version 1.4 http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#_4 http://www.w3.org/ns/prov#value Epithelial chloride (Cl-) transport is achieved by the coordinated action of symporters such as the Na+-K+-2Cl- cotransporter (NKCC1) and chloride channels such as the cystic fibrosis transmembrane conductance regulator (CFTR). As a secretory tissue, mammary epithelial cells are obvious candidates for such mechanisms, but Cl- transport and its hormonal regulation have been poorly delineated in mammary epithelial cells. We determined whether the mammary epithelial cell line, HC11, transports chloride and whether this was regulated by PRL, a hormone known to stimulate ion transport. HC11 cells express both CFTR and NKCC1. Exposure to PRL or PGE1 increased Cl- transport in HC11 cells. This was inhibited by the NKCC1 blocker, furosemide, and by the Cl- channel inhibitor, diphenylamine 2-carboxylate. Dose and time course of PRL action indicate that PRL had maximal effect on Cl- transport at 1 microg/ml and at 10 min of stimulation. Examination of the signaling pathways suggests that the PRL effect on Cl- transport does not involve an increase in [Ca2+]i or MAP kinase activity. RT-PCR analyses indicate that HC11 cells express mRNA for Janus kinase 1 (JAK1), JAK2, and signal transducer and activator of transcription 5 (STAT5) but not for JAK3. PRL treatment of HC11 cells increased phosphorylation of STAT5. The JAK2 inhibitor AG490 blocked phosphorylation of STAT5 and PRL-induced, but not PGE1-induced, Cl- transport. NKCC1, but not CFTR, is tyrosine phosphorylated in HC11 cells. PRL enhanced tyrosine phosphorylation of NKCC1, and this effect was attenuated by the JAK2 inhibitor AG490. These results are the first demonstrations of a role for tyrosine phosphorylation of NKCC1 and of the PRL-JAK2 cascade in the regulation of Cl- transport. http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#_4 http://www.w3.org/ns/prov#wasQuotedFrom http://www.ncbi.nlm.nih.gov/pubmed/11117534 http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#_5 http://www.w3.org/2000/01/rdf-schema#label Selventa http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#assertion http://www.w3.org/ns/prov#hadPrimarySource http://www.ncbi.nlm.nih.gov/pubmed/11117534 http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://resource.belframework.org/belframework/1.0/knowledge/large_corpus.bel http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#assertion http://www.w3.org/ns/prov#wasDerivedFrom http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#_4 http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk#pubinfo http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk http://purl.org/dc/terms/created 2014-07-03T14:29:53.610+02:00 http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk http://purl.org/pav/createdBy http://orcid.org/0000-0001-6818-334X http://www.tkuhn.ch/bel2nanopub/RAxVcfSIhBLmy5ydhip2BKIhANrLcbfcxu12L0h1yy4xk http://purl.org/pav/createdBy http://orcid.org/0000-0002-1267-0234