Antagonists of epithelial chloride channels inhibit cAMP synthesis.
In past studies we observed that the chloride channel blocker, diphenylamine-2-carboxylate (DPC) and chemically related drugs (Hoechst compounds 131, 143, 144) inhibited cAMP formation in mouse pituitary tumor cells. The object of this study was to determine whether these drugs inhibited chloride transport in human T-84 colonic carcinoma cells through an effect on cAMP metabolism. Chloride secretion (measured as 125I efflux from isotope-preloaded cells) was stimulated in a concentration-dependent manner by vasoactive intestinal polypeptide (VIP) (EC50 = 1.5 x 10(-10) M) which similarly increased cAMP synthesis (EC50 = 1.6 x 10(-8) M). The cAMP response to VIP was inhibited 17, 52, 55, and 78% maximally by DPC and compounds 144, 143, and 131, respectively. In untreated T-84 cells, 125I secretion fell by 66% after 3 min; VIP (10(-7) M) increased secretion about fivefold over the same period. Both basal and VIP-stimulated 125I secretion were inhibited up to 60% by compound 131. Pretreatment of cells with pertussis toxin did not attenuate the inhibitory effect of channel blockers on either VIP-stimulated cAMP synthesis or 125I secretion. The cationophore, A-23187, which had no effect on cAMP formation, and 8-Br-cAMP both stimulated 125I secretion from T-84 cells. These secretory responses were inhibited by compound 131. The mechanism by which phenylanthranilic acids antagonize cAMP synthesis and its significance is not known; however, the data suggest that this family of drugs may inhibit chloride transport by both cAMP-dependent and independent mechanisms.
Heisler
S
S
Department of Medicine, McGill University, Montréal, Qué., Canada.
D018118:Chloride Channels; D002712:Chlorides / Q000378:metabolism / Q000502:physiology; D003110:Colonic Neoplasms / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis*; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007455:Iodine / Q000378:metabolism / Q000502:physiology; D007457:Iodine Radioisotopes; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D037342:Pertussis Toxin; D013268:Stimulation, Chemical; D014407:Tumor Cells, Cultured; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D062367:ortho-Aminobenzoates / Q000494:pharmacology
D018118:Chloride Channels; D002712:Chlorides; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010566:Virulence Factors, Bordetella; D062367:ortho-Aminobenzoates; D014660:Vasoactive Intestinal Peptide; C001209:fenamic acid; D007455:Iodine; D000242:Cyclic AMP; D037342:Pertussis Toxin