PAR1-type thrombin receptor stimulates migration and matrix adhesion of human colon carcinoma cells by a PKCepsilon-dependent mechanism.
The proteinase-activated receptor1 (PAR1) was characterized as a functional receptor for thrombin in cells from different tumor entities. In colon carcinoma, its function has to be defined. In this study we demonstrate that the PAR1-selective agonist peptide TFLLRN induced activation of protein kinase C isoenzymes alpha and epsilon in human HT-29 colon carcinoma cells expressing PAR1 endogeneously. On the cellular level, TFLLRN and thrombin prompted HT-29 cell migration and matrix adhesion by a PKCepsilon-dependent mechanism as concluded because of the inhibition of PAR1-mediated effects by the PKC inhibitor bisindolylmaleimide I and the PKCepsilon translocation inhibitory peptide EAVSLKPT but not by the PKC inhibitor Gö 6976. In addition, blockade of PAR1 by RWJ 56110, a selective PAR1 antagonist, fully abolished the effect of thrombin on HT-29 cell migration and adhesion. Therefore, PAR1 seems to be the responsible receptor for thrombin-induced migration and adhesion of human colon carcinoma cells including PKCepsilon as an essential signal transducer.
Heider
Ilonka
I
Department of General and Visceral Surgery, Medical Faculty at the Friedrich Schiller University Jena, Erlanger Allee 101, D-07747, Germany.
Schulze
Beate
B
Oswald
Elke
E
Henklein
Peter
P
Scheele
Johannes
J
Kaufmann
Roland
R
D015153:Blotting, Western; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms; D011493:Protein Kinase C / Q000378:metabolism; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D012333:RNA, Messenger / Q000378:metabolism; D044463:Receptor, PAR-1 / Q000737:chemistry / Q000502:physiology*; D018179:Receptors, Thrombin / Q000378:metabolism / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013917:Thrombin / Q000378:metabolism; D013997:Time Factors
D007211:Indoles; D008301:Maleimides; D010455:Peptides; D020033:Protein Isoforms; D012333:RNA, Messenger; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin; C496628:PRKCA protein, human; C497255:PRKCE protein, human; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D013917:Thrombin; C070515:bisindolylmaleimide I