Abstract
As a first step in our efforts to delineate the role of CFTR in cellular phenotypes we have studied its expression in cultured human intestinal epithelial cells. In particular we have examined the effect of cellular differentiation on CFTR gene expression. CFTR mRNA was measured by quantitative densitometry of Northern blots and normalized to the amounts of pyruvate dehydrogenase message. We have found that in T84 cells the levels of CFTR mRNA do not change as the cells grow to confluence. In contrast, levels of CFTR mRNA increase by a factor of 10–20 as Caco2 cells grow after subculture. This change in the levels of CFTR mRNA is correlated with the morphological differentiation that occurs in Caco2 cells during culture. The potential significance of this observation is discussed.
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References
Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J., Rutter, W.J., 1979, Isolation of biologically active ribonucleic acid from sources enriched in ribonucleases,Biochemistry, 18:5294.
Dharmsathaphorn, K., Mandel, K.G., McRoberts, J.A., Tisdale, L.D. and Masui, H., 1984, A human colonic tumor cell line that maintains vectorial electrolyte transport, Am. J. Physiol., 246:G204.
Dharmasathaphorn, K., Mandel, K.G., Masui, H., and McRoberts, J.A., 1985, J. Clin. Invest., 75:462.
Feinberg, A.P. and Vogelstein, B., 1983, A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity, Anal. Biochem., 132:6.
Fourney, R.M., Miyakoshi, J., Day, R.S., and Paterson, M.C., 1988, Northern blotting: efficient RNA staining and transfer, Focus, 10:5.
Frizzell, R.A., Rechkemmer, G., and Shoemaker, R.L., 1986, Altered regulation of airway epithelial cell chloride channels in cystic fibrosis, Science, 23 3:558.
Grasset, E., Pinto, M., Dussaulx, E., Zweibaum, A., and Desjeux, J.-F., 1984, Epithelial properties of human colonic carcinoma cell line Caco-2: electrical parameters, Am. J.Physiol., 247:C260.
Grasset, E., Bemabeu, J., and Pinto, M., 1985, Epithelial properties of human colonic carcinoma cell line Caco-2: effect of secretagogues, Am. J.Physiol., 248:C410.
Knowles, M., Gatzy, J., and Boucher, R., 1981, Increased bioelectric potential difference across respiratory epithelia in cystic fibrosis, New England J. Med. 305:1489.
Li, M., McCann, J.D., Liedke, C.M., Naim, A.C., Greengard, P., and Welsh, M.J., 1988, Cyclic AMP-dependent protein kinase opens chloride channels in normal but not in cystic fibrosis airway epithelium, Nature, 331:358.
Pinto M., Robine-Leon, S., Appay, M.-D., Kedinger, M., Triadou, N., Dussaulx, E., Lacroix, B., Simon-Assmann, P., Haffen, K., Fogh, J., Zweibaum, A., 1983, Enterocyte differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture, Biol. Cell, 47:323.
Quinton, P.M., 1983, Chloride impermeability in cystic fibrosis, Nature, 301:421.
Quinton, P., 1989, Defective epithelial ion transport in cystic fibrosis, Clin. Chem., 35:726.
Riordan, J.R., Rommens, J.M., Kerem, B.-S., Alon, N., Rozmahel, R., Grzelczak, Z., Zielenski, J., Lok, S., Plavsic, N., Chou. J.-L., Drumm, M.L., Iannuzzi, M.C., Collins, F.S., Tsui, L.-C., 1989, Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA, Science, 245:1066.
Rousset, M., 1986, The human colon carcinoma cell lines HT-29 and Caco-2: two in vitro models for the study of intestinal differentiation, Biochimie, 68:1035.
Sato, K. and Sato, F., 1984, Defective beta-adrenergic response of cystic fibrosis sweat glands in vivo and in vitro, J. Clin.Invest., 73:1763.
Schoumacher, R.A., Shoemaker, R.L., Halm, D.R., Tallant, E.A., Wallace, R.W., and Frizzell, R.A., 1987, Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells, Nature, 3 30:752.
Widdicombe, J.H., Welsh, M.J., and Finkbeiner, W.E., 1985, Cystic fibrosis decreases the apical membrane chloride permeability of monolayers cultured from cells of tracheal epithelium. Proc. Natl. Aca. Sci. USA 82: 6167.
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Buchwald, M., Sood, R., Auerbach, W. (1991). Regulation of Expression of CFTR in Human Intestinal Epithelial Cells. In: Tsui, LC., Romeo, G., Greger, R., Gorini, S. (eds) The Identification of the CF (Cystic Fibrosis) Gene. Advances in Experimental Medicine and Biology, vol 290. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5934-0_24
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DOI: https://doi.org/10.1007/978-1-4684-5934-0_24
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