Lovastatin inhibits cAMP- and calcium-stimulated chloride secretion by T84 cells

Abstract

Isoprenylated proteins function in the processes of signal transduction and membrane vesicle trafficking. To investigate the role of isoprenylated proteins in secretagogue-stimulated epithelial ion transport, we studied the effects of lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, on adenosine 3',5'-cyclic monophosphate (cAMP)- and Ca(2+)-stimulated Cl- secretion by monolayers of T84 colonic epithelial cells. Lovastatin reduces protein isoprenylation in many cell types. In T84 cells, lovastatin reversibly inhibits forskolin-stimulated equivalent short-circuit current (I(sc)eq) by 50% after 2 days of treatment. The concentration of lovastatin resulting in half-maximal effects on forskolin-stimulated I(sc)eq is consistent with inhibition of protein isoprenylation, and lovastatin effects on forskolin-stimulated I(sc)eq are not associated with inhibition of cholesterol or glycoprotein biosynthesis. Lovastatin blocks N6,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate- and ionomycin-stimulated Isc, suggesting that it inhibits a process beyond the stimulation of cAMP and Ca2+ second-messenger systems. In monolayers in which the basolateral membrane has been permeabilized with nystatin, lovastatin inhibits cAMP activation of a diphenylamine-2-carboxylate-sensitive, apical membrane Cl- conductance. Our results are consistent with the hypothesis that an isoprenylated protein is involved in the regulation of a secretagogue-activated apical membrane Cl- conductance in T84 cells.