Ethanol increases the paracellular permeability of monolayers of CAPAN-1 pancreatic duct cells.
When grown on permeable supports, pancreatic duct adenocarcinoma CAPAN-1 cells establish very high values of transepithelial resistance (TER). The addition of ethanol produced a dose-related, reversible drop in the TER of these cells, ranging from 15% (with 1% ethanol) to 65% (with 10% ethanol). The ethanol effect was rapid and reversible. The resistance decrease was associated with an increase in monolayer permeability to mannitol. No significant decrease in cell ATP was detected for ethanol concentrations lower than 7%. Confocal vertical sections of calcein-loaded monolayers of CAPAN-1 cells, grown on plasticware, showed a progressive deflation of domes detectable after 5 min of treatment with 2% ethanol. Incubation in an ethanol-free medium caused a progressive dome restoration. Immunocytochemical analysis of ethanol-treated cells indicated that ZO-1 and occludin exhibited clear cut distribution changes while the perijunctional actin pattern was slightly modified. Electron microscopy showed that a discrete intercellular space was detectable between adjacent ethanol-treated cells but not between control cells. These data indicate that ethanol is a tight junction barrier opener in pancreatic duct cells.
Rotoli
B M
BM
General and Clinical Pathology Unit, Department of Experimental Medicine, Università degli Studi di Parma, 43100 Parma, Italy.
Orlandini
G
G
Guizzardi
S
S
Uggeri
J
J
Dall'Asta
V
V
Gazzola
G C
GC
Bussolati
O
O
Gatti
R
R
D000199:Actins / Q000378:metabolism; D021441:Carcinoma, Pancreatic Ductal / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D045744:Cell Line, Tumor; D000431:Ethanol / Q000494:pharmacology*; D006801:Humans; D007150:Immunohistochemistry; D008353:Mannitol / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D062793:Occludin; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D010539:Permeability / Q000187:drug effects; D010750:Phosphoproteins / Q000378:metabolism; D012997:Solvents / Q000494:pharmacology*; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein
D000199:Actins; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D010750:Phosphoproteins; D012997:Solvents; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D000431:Ethanol; D008353:Mannitol