Ion Transport in HT29 Colonic Carcinoma Cells

  • Karl Kunzelmann
  • Monika Tilmann
  • Rainer Greger
Part of the Advances in Comparative and Environmental Physiology book series (COMPARATIVE, volume 16)


The permanent cell line HT29 was established by Fogh and Trempe in 1975 from an adenocarcinoma of human colon (Fogh and Trempe 1975). In the following years, the cell line was examined and characterized with respect to many different biochemical and biophysical properties. HT29 cells, besides other colonic carcinoma cell lines such as T84 and CACO-2, serve as a very useful model in studying growth and differentiation of epithelial cells (Rousset 1986; Le Bivic et al. 1988; Hekmati et al. 1990). Moreover, the presence of a number of different hormone receptors was demonstrated in the membranes of HT29 cells. Second messenger systems activated by receptor binding were examined and are still under investigation (cf. Table 1). In all instances, the occupation of the receptor was linked to a change in the ion conductance properties of the HT29 cell. In terms of membrane transport proteins, in addition to the (Na+/K+)-ATPase, the presence of the following systems has been shown in HT29 cells: the Na+/H+ exchanger (Cantiello and Lanier 1989); the Na+/2Cl/K+ cotransporter (Kim et al. 1988); a K+ conductance (Wu et al. 1991); and a Cl conductance (Ziss et al. 1987; Lohrmann et al. 1991; Kunzelmann et al. 1992a). Because of their polar growth, which is developed under certain culture conditions, HT29 cells are also useful for studying electrolyte transport, and they are commonly looked at as a paradigm of the colonic crypt cell.


Cystic Fibrosis HT29 Cell Cystic Fibrosis Transmembrane Conductance Regulator Chloride Channel Regulatory Volume Decrease 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Amar S, Kitabgi P, Vincent J-P (1986) Activation of phosphatidylinositol turnover by neurotensin receptors in the human colonic adenocarcinoma cell line HT29. FEBS Lett 201: 31–36PubMedCrossRefGoogle Scholar
  2. Anderson MP, Welsh MJ (1990) Fatty acids inhibit apical membrane chloride channels in airway epithelia. Proc Natl Acad Sci USA 87: 7334–7338PubMedCentralPubMedCrossRefGoogle Scholar
  3. Augeron C, Laboisse CL (1984) Emergence of permanently differentiated cell clones in a human colonic cancer cell line in culture after treatment with sodium butyrate. Cancer Res 44: 3961–3969PubMedGoogle Scholar
  4. Bouscarel B, Cortinovis C, Carpene C, Murat JC, Paris H (1985) a2-Adrenoreceptors in the HT29 human colon adenocarcinoma cell line: characterization with [3H] clonidine; effects on cyclic AMP accumulation. Eur J Pharmacol 107: 223–231CrossRefGoogle Scholar
  5. Bozou J-C, Rochet N, Magnaldo I, Vincent J-P, Kitabgi P (1989) Neurotensin stimulates inositol trisphosphate-mediated calcium mobilization but not protein kinase C activation in HT29 cells. Biochem J 264: 871–878PubMedCentralPubMedGoogle Scholar
  6. Cantiello HF, Lanier SM (1989) a2-Adrenergic receptors and the Na/H exchanger in the intestinal epithelial cell line HT-29. J Biol Chem 264: 16000–16007PubMedGoogle Scholar
  7. Fogh J, Trempe G (1975) New human tumor cell lines. In: Fogh J (ed) Human tumor cells in vitro. Plenum Press, New York, pp 115–141CrossRefGoogle Scholar
  8. Franklin CC, Turner JT, Kim HD (1989) Regulation of Na+/K+/Cl- cotransport and [3H]bumetanide binding site density by phorbol esters in HT29 cells. J Biol Chem 264: 6667–6673PubMedGoogle Scholar
  9. Frizzell RA, Schoumacher RA, Shoemaker RL, Halm DR (1987) Chloride channel regulation in secretory epithelial cells. Pediatr Pulmonol Suppl 1: 24–25Google Scholar
  10. Gluck D, Kelly S, Al-Awqati Q (1982) The proton translocating ATPase responsible for urinary acidification. J Biol Chem 257 (16): 9230–9233PubMedGoogle Scholar
  11. Gögelein H, Greger R (1987) Properties of single K+ channels in the basolateral membrane of rabbit proximal straight tubules. Pflügers Arch Eur J Physiol 410: 288–295CrossRefGoogle Scholar
  12. Greger R, Kunzelmann K (1991) Simultaneous recording of the cell membrane potential and properties of the cell attached membrane of HT29 colon carcinoma and CF-PAC cells. Pflügers Arch Eur J Physiol 419: 209–211CrossRefGoogle Scholar
  13. Greger R, Schlatter E (1984) Mechanism of NaCl secretion in the rectal gland of spiny dogfish (Squalus acanthias). I. Experiments in isolated in vitro perfused rectal gland tubules. Pflügers Arch Eur J Physiol 402: 63–75CrossRefGoogle Scholar
  14. Greger R, Gögelein H, Schlatter E (1987) Potassium channels in the basolateral membrane of the rectal gland of the dogfish (Squalus acanthias). Pflügers Arch Eur J Physiol 409: 100–106CrossRefGoogle Scholar
  15. Hansen CP, Roch B, Kunzelmann K, Greger R (1991) Inhibition of epithelial chloride channels by epithelial cytosols. Pflügers Arch Eur J Physiol 419: R101CrossRefGoogle Scholar
  16. Hayslett JP, Gögelein H, Kunzelmann K, Greger R (1987) Characteristics of apical chloride channels in human colon cells (HT29). Pflügers Arch Eur J Physiol 410: 487–494CrossRefGoogle Scholar
  17. Hekmati M, Polak-Charcon S, Ben-Shaul Y (1990) A morphological study of a human adenocarcinoma cell line (HT29) differentiating in culture. Similarities to intestinal embryonic development. Cell Differ Dev 31: 207–218PubMedCrossRefGoogle Scholar
  18. Huet C, Sahuquillo-Merino C, Coudrier E, Louvard D (1987) Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation. J Cell Biol 105: 345–357PubMedCrossRefGoogle Scholar
  19. Kim HD, Tsai Y-S, Franklin CC, Turner JT (1988) Characterization of Na+-K+-Clcotransport in cultured HT29 human colonic adenocarcinoma cells. Biochim Biophys Acta 946: 397–404PubMedCrossRefGoogle Scholar
  20. Kreusel KM, Fromm M, Lempart U, Sorgenfrei D, Hegel U (1990) Epithelial monolayers of differentiated HT-29 cells exhibit crypt-like chloride secretion. Pflügers Arch Eur J Physiol 415: R33 (Abstr)Google Scholar
  21. Krick W, Disser J, Hazama A, Burckhardt G, Frömter E (1991) Evidence for a cytosolic inhibitor of epithelial chloride channels. Pflügers Arch Eur J Physiol 418: 491–499CrossRefGoogle Scholar
  22. Kunzelmann K, Pavenstädt H, Beck C, finalÖ, Emmrich P, Arndt Hi, Greger R (1989a) Characterization of potassium channels in respiratory cells I. General properties. Pflügers Arch Eur J Physiol 414: 291–296CrossRefGoogle Scholar
  23. Kunzelmann K, Pavenstädt H, Greger R (1989b) Characterization of potassium channels in respiratory cells II. Inhibitors and regulation. Pflügers Arch Eur J Physiol 414: 297–303CrossRefGoogle Scholar
  24. Kunzelmann K, Pavenstädt H, Greger R (1989c) Properties and regulation of chloride channels in cystic fibrosis and normal airway epithelial cells. Pflügers Arch Eur J Physiol 415: 172–182CrossRefGoogle Scholar
  25. Kunzelmann K, Gerlach L, Fröbe U, Greger R (1991a) Bicarbonate permeability of epithelial chloride channels. Pflügers Arch Eur J Physiol 417: 616–621CrossRefGoogle Scholar
  26. Kunzelmann K, Tilmann M, Hansen CP, Greger R (1991b) Inhibition of epithelial chloride channels by cytosol. Pflügers Arch Eur J Physiol 418: 479–490CrossRefGoogle Scholar
  27. Kunzelmann K, Grolik M, Kubitz R, Greger R (1992a) cAMP dependent activation of small conductance Cl-channels in HT29 colon carcinoma cells. Pflügers Arch Eur J Physiol (in press)Google Scholar
  28. Kunzelmann K, Kubitz R, Grolik M, Warth R, Greger R (1992b) Small conductance CI-channels in HT29 cells: activation by Ca2+, hypotonic cell swelling and 8-Br-cGMP. Pflügers Arch Eur J Physiol (in press)Google Scholar
  29. Laburthe M, Rousset M, Boissard C, Zweibaum A, Rosselin G (1978) Vasoactive intestinal peptide: a potent stimulator of adenosine 3′: 5′-cyclic monophophate accumulation in gut carcinoma cell lines in culture. Proc Natl Acad Sci USA 75: 2772–2775PubMedCentralPubMedCrossRefGoogle Scholar
  30. Le Bivic A, Hirn M, Reggio H (1988) HT-29 cells are an in vitro model for the generation of cell polarity in epithelia during embryonic differentiation. Proc Natl Acad Sci USA 85: 136–140PubMedCentralPubMedCrossRefGoogle Scholar
  31. Li M, McCann JD, Liedtke CM, Nairn AC, Greengard P, Welsh MJ (1988) Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium. Nature 331: 358–360PubMedCrossRefGoogle Scholar
  32. Lohrmann E, Cabantchik ZI, Greger R (1991) The membrane potential (PD) and the conductance properties of HT29 Cells. Pflügers Arch Eur J Physiol 418: R326 (Abstr)Google Scholar
  33. Montrose MH (1990) Linkage between second messenger levels and cell volume in cultured intestinal cells (HT29–C1). FASEB J 4: A448 (Abstr)Google Scholar
  34. Montrose MH, Murer H (1989) Polarized response to propionate in cultured human colonic epithelial cells (HT29/C1). Organization of membrane polarity in epithelial cells, Arolla (Abstr )Google Scholar
  35. Montrose-Rafizadeh C, Guggion WB, Montrose MH (1990) Expression of CFTR is modulated by cell differentiation in a cloned human intestinal cell line. Pediatr Pulmonol Suppl 5: 192 (Abstr)Google Scholar
  36. Montrose-Rafizadeh C, Guggino WB, Montrose MH (1991) Cellular differentiation regulates expression of CI transport and cystic fibrosis transmembrane conductance regulator mRNA in human intestinal cells. J Biol Chem 266: 4495–4499PubMedGoogle Scholar
  37. Nitschke R, Leipziger J, Greger R (1993a) Intracellular Ca2+ transients in HT29 cells induced by hypotonic cell swelling. Pflügers Arch (in press)Google Scholar
  38. Nitschke R, Leipziger J, Greger R (1993b) Agonist induced intracellular Ca2+ transients in HT29 cells. Pflügers Arch (in press)Google Scholar
  39. Paris H, Bouscarel B, Cortinovis C, Murat JC (1985) Growth-related variation of a2adrenergic receptivity in the HT29 adenocarcinoma cell-line from human colon. FEBS Lett 184: 82–86PubMedCrossRefGoogle Scholar
  40. Rechkemmer GR, Bökenkamp D, Jeromin A (1991) Intracellular calcium concentration in the human colonic tumor cell line HT-CL19A. Effects of carbachol, forskolin and vasoactive intestinal polypeptide. Pflügers Arch Eur J Physiol 418: R59 (Abstr)Google Scholar
  41. Remy L, Marvaldi J, Rua S, Secchi J, Lechene de la Porte P (1984) The role of intracellular lumina in the repolarization process of a colonic adenocarcinoma cell line. Virchows Arch 46: 297–305CrossRefGoogle Scholar
  42. Reynier M, Sari H, d’Anglebermes M, Ah Kye E, Pasero L (1991) Differences in lipid characteristics of undifferentiated and entrocytic-differentiated HT29 human colon cells. Cancer Res 51: 1270–1277PubMedGoogle Scholar
  43. Rommens JM, Iannuzzi B-SK, Drumm ML, Melmer G, Dean M, Rozmahel R, Cole JL, Kennedy D, Hidaka N, Zsiga M, Buchwald M, Riordan JR, Tsui L-C, Collins FS (1989) Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245: 1059–1065PubMedCrossRefGoogle Scholar
  44. Rothenberg P, Glaser L, Schlesinger P, Cassel D (1983) Activation of Na+/H+ exchange by epidermal growth factor elevates intracellular pH in A 431 cells. J Biol Chem 258: 12644–12653PubMedGoogle Scholar
  45. Rotin D, Steele-Norwood D, Grinstein S, Tannock I (1989) Requirement of the Na+/HT exchanger for tumor growth. Cancer Res 49: 205–211PubMedGoogle Scholar
  46. 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: 1035–1040PubMedCrossRefGoogle Scholar
  47. Schoumacher RA, Shoemaker RL, Halm DR, Tallant EA, Wallace RW, Frizzell RA (1987) Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells. Nature 330: 752–754PubMedCrossRefGoogle Scholar
  48. Simon-Assman P, Bouziges F, Daviaud D, Haffen K, Kedinger M (1987) Synthesis of glucosaminoglycans by undifferentiated and differentiated HT29 human colonic cancer cells. Cancer Res 47: 4478–4484Google Scholar
  49. Sood R, Auerbach W, Shannon W, Buchwald M (1990) Regulation of expression of CFTR with in vitro differentiation of human intestinal epithelial cells. Pediatr Pulmonol Suppl 5: 194 (Abstr)Google Scholar
  50. Szolgay-Daniel E, Carlsson J, Zierold K, Holtermann G, Dufau E, Acker H (1991) Effect of amiloride treatment on U-118 MG and U-251 MG human glioma and HT-29 human colon carcinoma cells. Cancer Res 51: 1039–1044PubMedGoogle Scholar
  51. Tilly BC, Kansen M, Gageldonk PGM, van den Berghe N, Galjaard H, Bijman J, De Jonge HR (1991) G-proteins mediate intestinal chloride channel activation. J Biol Chem 266: 2036–2040PubMedGoogle Scholar
  52. Tilmann M, Kunzelmann K, Fröbe U, Cabantchik I, Lang HJ, Englert HC, Greger R (1991) Different types of blockers of the intermediate conductance outwardly rectifying chloride channel (ICOR) in epithelia. Pflügers Arch Eur J Physiol 418: 556–563CrossRefGoogle Scholar
  53. Turner JT, Franklin CC, Bollinger DW, Kim HD (1990) Vasoactive intestinal peptide stimulates active K+ transport and Na+/K+/Cl-cotransport in HT-29 cells. Am J Physiol 258: C266 - C273PubMedGoogle Scholar
  54. Welsh MJ (1986) An apical-membrane chloride channel in human tracheal epithelium. Science 232: 1648–1649PubMedCrossRefGoogle Scholar
  55. Wu H, Franklin CC, Kim HD, Turner JT (1991) Regulation of calcium-activated potassium efflux by neurotensin and other agents in HT-29 cells. Am J Physiol 260: C35 - C42PubMedGoogle Scholar
  56. Ziss W, Hegel U (1988) Human colon cancer cells (HT-29) exhibit ion transport properties of intestinal crypt cells. Pflügers Arch Eur J Physiol 411: R81 (Abstr)Google Scholar
  57. Ziss W, Fromm M, Sorgenfrei D, Hegel U (1987) Effect of chloride step changes on the membrane potential in the human colonic carcinoma cancer cell line HT-29. Pflügers Arch Eur J Physiol 408: R32 (Abstr)Google Scholar
  58. Zweibaum A, Pinto M, Chevalier G, Dussaulx E, Triadou N, Lacroix B, Haffen K, Brun J-L, Rousset M (1985) Enterocytic differentiation of a subpopulation of the human tumor cell line HT-29 selected for growth in sugar-free medium and its inhibition by glucose. J Cell Physiol 122: 21–29PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1993

Authors and Affiliations

  • Karl Kunzelmann
  • Monika Tilmann
  • Rainer Greger
    • 1
  1. 1.Physiologisches InstitutAlbrecht-Ludwigs-UniversitätFreiburgGermany

Personalised recommendations