Transactivation of the epidermal growth factor receptor mediates cholinergic agonist-induced proliferation of H508 human colon cancer cells.
Some human colon cancer cell lines (e.g., H508 cells) express M3 subtype muscarinic receptors that are activated by cholinergic agonists. The objective of the present study was to determine the cellular mechanisms underlying M3 muscarinic receptor-mediated proliferation of H508 human colon cancer cells. In H508 cells, but not in SNU-C4 cells that do not express muscarinic receptors, acetylcholine stimulated calcium-dependent phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) and p90 ribosomal S6 kinase and consequent cell proliferation. Atropine or inhibitors of MAPK phosphorylation blocked these effects. Conversely, the actions of epidermal growth factor (EGF) on H508 cells were neither calcium dependent nor mediated by cholinergic mechanisms. Both acetylcholine- and EGF-induced phosphorylation of p44/42 MAPK was abolished in the presence of EGF receptor (EGFR) inhibitors (AG1478 and PD168393). In Chinese hamster ovary cells transfected with the rat M3 muscarinic receptor, which lack EGFR, acetylcholine-induced MAPK phosphorylation was not altered in the presence of EGFR inhibitors. In H508 cells, protein kinase C inhibitors did not alter acetylcholine- or EGF-induced MAPK phosphorylation. Finally, inhibition of EGFR activation abolished acetylcholine-induced H508 cell proliferation. These data indicate that, in H508 human colon cancer cells, cholinergic ligand interaction with M3 muscarinic receptors results in transactivation of EGFR, thereby stimulating cellular proliferation.
Division of Gastroenterology, VA Maryland Health Care System and Program in Oncology, University of Maryland Greenebaum Cancer Center, Baltimore, Maryland 21201, USA.
Kunrong
K
Cheng
Piotr
P
Zimniak
Jean-Pierre
JP
Raufman
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D018679:Cholinergic Agonists / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000201:enzymology / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006224:Cricetinae; D004347:Drug Interactions; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D018727:Muscarinic Antagonists / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011799:Quinazolines; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000378:metabolism; D015533:Transcriptional Activation / Q000502:physiology; D020032:Tyrphostins / Q000494:pharmacology
D018679:Cholinergic Agonists; D005419:Flavonoids; D018727:Muscarinic Antagonists; D011799:Quinazolines; D020032:Tyrphostins; C101044:RTKI cpd; D004533:Egtazic Acid; D001285:Atropine; D019869:Phosphatidylinositol 3-Kinases; D066246:ErbB Receptors; D038744:Ribosomal Protein S6 Kinases, 90-kDa; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D000109:Acetylcholine; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one