Migration of primary cultured rabbit gastric epithelial cells requires intact protein kinase C and Ca2+/calmodulin activity.
Superficial gastric mucosal injury is rapidly repaired by epithelial cell migration. This study aims to characterize the intracellular signal transduction pathways underlying the repair process. Primary monolayer cultures of rabbit gastric epithelial cells were wounded. The measured spontaneous cell migration speed at the edge of the wound was 457+/-89 microm/24 hr. Epidermal growth factor stimulated and genistein (receptor tyrosine protein kinase inhibitor) inhibited cell migration significantly. Down-regulation of protein Kinase C (PKC) with long-term phorbol 12-myristate 13-acsetate or inhibition with calphostin-C significantly inhibited cell migration. Blocking of Ca2+ channels with verapamil and endogenous Ca2+ release with TMB-8 or inhibition of the Ca2+/calmodulin complex with calmidazolium likewise significantly inhibited migration speed and also abolished the rise of [Ca2+]i, which was measured in migrating cells. Modulation of the cAMP-PKA pathway or prostaglandin synthesis had no influence on cell migration. Gastric epithelial cell migration implies activation of receptor tyrosine kinase. It is associated with increased [Ca2+]i and requires an intact Ca2+/calmodulin complex. Intact PKC activity also is needed.
Ranta-Knuuttila
Tuula
T
Department of Gastroenterologic Surgery, Helsinki University Central Hospital, Finland.
Kiviluoto
Tuula
T
Mustonen
Harri
H
Puolakkainen
Pauli
P
Watanabe
Sumio
S
Sato
Nobuhiro
N
Kivilaakso
Eero
E
D000818:Animals; D002118:Calcium / Q000502:physiology*; D015220:Calcium Channels / Q000187:drug effects; D002147:Calmodulin / Q000502:physiology*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D015536:Down-Regulation; D004847:Epithelial Cells / Q000502:physiology*; D005707:Gallic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology*; D007093:Imidazoles / Q000494:pharmacology; D008297:Male; D009281:Naphthalenes / Q000494:pharmacology; D011453:Prostaglandins / Q000096:biosynthesis; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D011817:Rabbits; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D015220:Calcium Channels; D002147:Calmodulin; D007093:Imidazoles; D009281:Naphthalenes; D011453:Prostaglandins; C031938:calmidazolium; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; D005707:Gallic Acid; D011493:Protein Kinase C; C058819:calphostin C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium