The formation of intracellular lumina with apical differentiation is observed in several cancerous epithelial cell lines including human hepatocarcinoma. This disorder of cell polarization can be induced by the inhibition of cell-cell communication, a known factor of carcinogenesis. This work was designed to study the effects of ethanol on the differentiation of hepatocytes in short-term culture. Isolated hepatocytes were plated on plastic culture dishes that were 35 mm in diameter (10(6) cells/dish). Three hours after plating, the hepatocytes were incubated in the presence of 20 mmol/L ethanol for 1 hr. Treated cells were compared with controls using morphometric methods after conventional treatment for ultramicroscopy and by measuring cellular dye coupling by the fluorescent Lucifer Yellow CH transfer method. Bile canaliculi formation decreased in alcohol-treated cells (6.5% vs. 9.9%, 2p less than 0.05), whereas intracellular lumina incidence increased (3.1% vs. 0.5%, 2p less than 0.01). In parallel, the dye-coupling capacity decreased significantly when hepatocytes were treated with alcohol (2p less than 0.01). This work shows that short-term ethanol treatment induces significant disturbances of cell polarization and inhibits the reestablishment of cell-cell communication in cultured hepatocytes. These disorders could, at least in part, explain the carcinogenic effects of ethanol.