The Journal of Membrane Biology

, Volume 240, Issue 1, pp 35–46 | Cite as

Characterization of Butyrate Uptake by Nontransformed Intestinal Epithelial Cell Lines

  • Pedro Gonçalves
  • João R. Araújo
  • Fátima Martel


Butyrate (BT) is one of the main end products of anaerobic bacterial fermentation of dietary fiber within the human colon. Among its recognized effects, BT inhibits colon carcinogenesis. Our aim was to characterize uptake of BT by two nontransformed intestinal epithelial cell lines: rat small intestinal epithelial (IEC-6) and fetal human colonic epithelial (FHC) cells. Uptake of 14C-BT by IEC-6 cells was (1) time- and concentration-dependent; (2) pH-dependent; (3) Na+-, Cl- and energy-dependent; (4) inhibited by BT structural analogues; (5) sensitive to monocarboxylate transporter 1 (MCT1) inhibitors; and (6) insensitive to DIDS and amiloride. IEC-6 cells express both MCT1 and Na+-coupled monocarboxylate transporter 1 (SMCT1) mRNA. We conclude that 14C-BT uptake by IEC-6 cells mainly involves MCT1, with a small contribution of SMCT1. Acute exposure to ethanol, acetaldehyde, indomethacin, resveratrol and quercetin reduced 14C-BT uptake. Chronic exposure to resveratrol and quercetin reduced 14C-BT uptake but had no effect on either MCT1 or SMCT1 mRNA levels. Uptake of 14C-BT by FHC cells was time- and concentration-dependent but pH-, Na+-, Cl- and energy-independent and insensitive to BT structural analogues and MCT1 inhibitors. Although MCT1 (but not SMCT1) mRNA expression was found in FHC cells, the characteristics of 14C-BT uptake by FHC cells did not support either MCT1 or SMCT1 involvement. In conclusion, uptake characteristics of 14C-BT differ between IEC-6 and FHC cells. IEC-6 cells demonstrate MCT1- and SMCT1-mediated transport, while FHC cells do not.


Butyrate uptake Nontransformed intestinal epithelial cell Monocarboxylate transporter type 1 Xenobiotics 



This work was supported by Fundação para a Ciência e a Tecnologia (FCT) and Programa Ciência, Tecnologia e Inovação do Quadro Comunitário de Apoio (PTDC/SAU-FCF/67805/2006). The authors thank M. J. Pinho (Institute of Pharmacology and Therapeutics, Faculty of Medicine of Porto, Portugal) for her help concerning real-time qRT-PCR.


  1. Alrefai WA, Tyagi S, Gill R, Saksena S, Hadjiagapiou C, Mansour F et al (2004) Regulation of butyrate uptake in Caco-2 cells by phorbol 12-myristate 13-acetate. Am J Physiol Gastrointest Liver Physiol 286:G197–G203PubMedCrossRefGoogle Scholar
  2. Ananth S, Zhuang L, Gopal E, Itagaki S, Ellappan B, Smith SB et al (2010) Diclofenac-induced stimulation of SMCT1 (SLC5A8) in a heterologous expression system: a RPE specific phenomenon. Biochem Biophys Res Commun 394:75–80PubMedCrossRefGoogle Scholar
  3. Anzai N, Kanai Y, Endou H (2006) Organic anion transporter family: current knowledge. J Pharmacol Sci 100:411–426PubMedCrossRefGoogle Scholar
  4. Araújo JR, Gonçalves P, Martel F (2008) Modulation of glucose uptake in a human trophoblast cell line (BeWo) by dietary bioactive compounds and drugs of abuse. J Biochem 144:177–186PubMedCrossRefGoogle Scholar
  5. Bongaerts BW, de Goeij AF, van den Brandt PA, Weijenberg MP (2006) Alcohol and the risk of colon and rectal cancer with mutations in the K-ras gene. Alcohol 38:147–154PubMedCrossRefGoogle Scholar
  6. Borthakur A, Gill RK, Hodges K, Ramaswamy K, Hecht G, Dudeja PK (2006) Enteropathogenic Escherichia coli inhibits butyrate uptake in Caco-2 cells by altering the apical membrane MCT1 level. Am J Physiol Gastrointest Liver Physiol 290:G30–G35PubMedCrossRefGoogle Scholar
  7. Borthakur A, Anbazhagan AN, Kumar A, Raheja G, Singh V, Ramaswamy K et al (2010) The probiotic Lactobacillus plantarum counteracts TNF-α-induced downregulation of SMCT1 expression and function. Am J Physiol Gastrointest Liver Physiol 299:G928–G934PubMedCrossRefGoogle Scholar
  8. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  9. Buyse M, Sitaraman SV, Liu X, Bado A, Merlin D (2002) Luminal leptin enhances CD147/MCT-1-mediated uptake of butyrate in the human intestinal cell line Caco2-BBE. J Biol Chem 277:28182–28190PubMedCrossRefGoogle Scholar
  10. Choi JS, Jin MJ, Han HK (2005) Role of monocarboxylic acid transporters in the cellular uptake of NSAIDs. J Pharm Pharmacol 57:1185–1189PubMedCrossRefGoogle Scholar
  11. Coady MJ, Chang MH, Charron FM, Plata C, Wallendorff B, Sah JF et al (2004) The human tumor suppressor gene SLC5A8 expresses a Na+-monocarboxylate cotransporter. J Physiol 557:719–731PubMedCrossRefGoogle Scholar
  12. Cresci GA, Thangaraju M, Mellinger JD, Liu K, Ganapathy V (2010) Colonic gene expression in conventional and germ-free mice with a focus on the butyrate receptor GPR109A and the butyrate transporter SLC5A8. J Gastrointest Surg 14:449–461PubMedCrossRefGoogle Scholar
  13. Cuff MA, Shirazi-Beechey SP (2004) The importance of butyrate transport to the regulation of gene expression in the colonic epithelium. Biochem Soc Trans 32:1100–1102PubMedCrossRefGoogle Scholar
  14. Cuff MA, Lambert DW, Shirazi-Beechey SP (2002) Substrate-induced regulation of the human colonic monocarboxylate transporter, MCT1. J Physiol 539:361–371PubMedCrossRefGoogle Scholar
  15. Cuff M, Dyer J, Jones M, Shirazi-Beechey S (2005) The human colonic monocarboxylate transporter isoform 1: its potential importance to colonic tissue homeostasis. Gastroenterology 128:676–686PubMedCrossRefGoogle Scholar
  16. Elwood PC, Gallagher AM, Duthie GG, Mur LA, Morgan G (2009) Aspirin, salicylates, and cancer. Lancet 373:1301–1309PubMedCrossRefGoogle Scholar
  17. Fraga S, Serrão MP, Soares-da-Silva P (2002) l-type amino acid transporters in two intestinal epithelial cell lines function as exchangers with neutral amino acids. J Nutr 132:733–738PubMedGoogle Scholar
  18. Fujita I, Akagi Y, Hirano J, Nakanishi T, Itoh N, Muto N et al (2000) Distinct mechanisms of transport of ascorbic acid and dehydroascorbic acid in intestinal epithelial cells (IEC-6). Res Commun Mol Pathol Pharmacol 107:219–231PubMedGoogle Scholar
  19. Ganapathy V, Thangaraju M, Gopal E, Martin PM, Itagaki S, Miyauchi S et al (2008) Sodium-coupled monocarboxylate transporters in normal tissues and in cancer. AAPS J 10:193–199PubMedCrossRefGoogle Scholar
  20. Gonçalves P, Araújo JR, Pinho MJ, Martel F (2009) Modulation of butyrate transport in Caco-2 cells. Naunyn-Schmiedeberg Arch Pharmacol 379:325–336CrossRefGoogle Scholar
  21. Gonçalves P, Araújo JR, Pinho MJ, Martel F (in press) In vitro studies on the inhibition of colon cancer by butyrate and polyphenolic compounds. Nutr CancerGoogle Scholar
  22. Gupta N, Martin PM, Prasad PD, Ganapathy V (2006) SLC5A8 (SMCT1)-mediated transport of butyrate forms the basis for the tumor suppressive function of the transporter. Life Sci 78:2419–2425PubMedCrossRefGoogle Scholar
  23. Hadjiagapiou C, Schmidt L, Dudeja PK, Layden TJ, Ramaswamy K (2000) Mechanism(s) of butyrate transport in Caco-2 cells: role of monocarboxylate transporter 1. Am J Physiol Gastrointest Liver Physiol 279:G775–G780PubMedGoogle Scholar
  24. Halestrap AP, Meredith D (2004) The SLC16 gene family—from monocarboxylate transporters (MCTs) to aromatic amino acid transporters and beyond. Pflugers Arch 447:619–628PubMedCrossRefGoogle Scholar
  25. Hamer HM, Jonkers D, Venema K, Vanhoutvin S, Troost FJ, Brummer RJ (2008) The role of butyrate on colonic function. Aliment Pharmacol Ther 27:104–119PubMedCrossRefGoogle Scholar
  26. Harig JM, Ng EK, Dudeja PK, Brasitus TA, Ramaswamy K (1996) Transport of n-butyrate into human colonic luminal membrane vesicles. Am J Physiol Gastrointest Liver Physiol 271:G415–G422Google Scholar
  27. Inui K, Quaroni A, Tillotson LG, Isselbacher KJ (1980) Amino acid and hexose transport by cultured crypt cells from rat small intestine. Am J Physiol 239:C190–C196PubMedGoogle Scholar
  28. Itagaki S, Gopal E, Zhuang L, Fei YJ, Miyauchi S, Prasad PD et al (2006) Interaction of ibuprofen and other structurally related NSAIDs with the sodium-coupled monocarboxylate transporter SMCT1 (SLC5A8). Pharm Res 23:1209–1216PubMedCrossRefGoogle Scholar
  29. Jakobs ES, Paterson AR (1986) Sodium-dependent, concentrative nucleoside transport in cultured intestinal epithelial cells. Biochem Biophys Res Commun 140:1028–1035PubMedCrossRefGoogle Scholar
  30. Kirk P, Wilson MC, Heddle C, Brown MH, Barclay AN, Halestrap AP (2000) CD147 is tightly associated with lactate transporters MCT1 and MCT4 and facilitates their cell surface expression. EMBO J 19:3896–3904PubMedCrossRefGoogle Scholar
  31. Konishi Y, Kobayashi S, Shimizu M (2003) Tea polyphenols inhibit the transport of dietary phenolic acids mediated by the monocarboxylic acid transporter (MCT) in intestinal Caco-2 cell monolayers. J Agric Food Chem 51:7296–7302PubMedCrossRefGoogle Scholar
  32. Lecona E, Olmo N, Turnay J, Santiago-Gómez A, López de Silanes I, Gorospe M et al (2008) Kinetic analysis of butyrate transport in human colon adenocarcinoma cells reveals two different carrier-mediated mechanisms. Biochem J 409:311–320PubMedCrossRefGoogle Scholar
  33. Martínez ME, Marshall JR, Giovannucci E (2008) Diet and cancer prevention: the roles of observation and experimentation. Nat Rev Cancer 8:694–703PubMedCrossRefGoogle Scholar
  34. Morris ME, Felmlee MA (2008) Overview of the proton-coupled MCT (SLC16A) family of transporters: characterization, function and role in the transport of the drug of abuse gamma-hydroxybutyric acid. AAPS J 10:311–321PubMedCrossRefGoogle Scholar
  35. Murota K, Matsui N, Kawada T, Takahashi N, Shintani T, Sasaki K et al (2001) Influence of fatty alcohol and other fatty acid derivatives on fatty acid uptake into rat intestinal epithelial cells. Lipids 36:21–26PubMedCrossRefGoogle Scholar
  36. Muzyka A, Tarkany O, Yelizanof V, Sergienko U, Boichuk A (2005) Non-linear regression (curve fit). GraphPad Prism for Windows, San DiegoGoogle Scholar
  37. Park Y, Hunter DJ, Spiegelman D, Bergkvist L, Berrino F, van den Brandt PA et al (2005) Dietary fiber intake and risk of colorectal cancer: a pooled analysis of prospective cohort studies. JAMA 294:2849–2857PubMedCrossRefGoogle Scholar
  38. Pöschl G, Seitz HK (2004) Alcohol and cancer. Alcohol Alcohol 39:155–165PubMedGoogle Scholar
  39. Quaroni A, Wands J, Trelstad RL, Isselbacher KJ (1979) Epithelioid cell cultures from rat small intestine: characterization by morphologic and immunologic criteria. J Cell Biol 80:248–265PubMedCrossRefGoogle Scholar
  40. Ritzhaupt A, Ellis A, Hosie KB, Shirazi-Beechey SP (1998) The characterization of butyrate transport across pig and human colonic luminal membrane. J Physiol 507:819–830PubMedCrossRefGoogle Scholar
  41. Said HM, Ma TY, Ortiz A, Tapia A, Valerio CK (1997) Intracellular regulation of intestinal folate uptake: studies with cultured IEC-6 epithelial cells. Am J Physiol Cell Physiol 272:C729–C736Google Scholar
  42. Saksena S, Dwivedi A, Gill RK, Singla A, Alrefai WA, Malakooti J et al (2009a) PKC-dependent stimulation of the human MCT1 promoter involves transcription factor AP2. Am J Physiol Gastrointest Liver Physiol 296:G275–G283PubMedCrossRefGoogle Scholar
  43. Saksena S, Theegala S, Bansal N, Gill RK, Tyagi S, Alrefai WA et al (2009b) Mechanisms underlying modulation of monocarboxylate transporter 1 (MCT1) by somatostatin in human intestinal epithelial cells. Am J Physiol Gastrointest Liver Physiol 297:G878–G885PubMedCrossRefGoogle Scholar
  44. Schröder O, Opritz J, Stein J (2000) Substrate and inhibitor specificity of butyrate uptake in apical membrane vesicles of the rat distal colon. Digestion 62:152–158PubMedCrossRefGoogle Scholar
  45. Seitz HK, Homann N (2007) The role of acetaldehyde in alcohol-associated cancer of the gastrointestinal tract. Novartis Found Symp 285:110–119PubMedCrossRefGoogle Scholar
  46. Shim CK, Cheon EP, Kang KW, Seo KS, Han HK (2007) Inhibition effect of flavonoids on monocarboxylate transporter 1 (MCT1) in Caco-2 cells. J Pharm Pharmacol 59:1515–1519PubMedCrossRefGoogle Scholar
  47. Siddiqui KM, Chopra DP (1984) Primary and long term epithelial cell cultures from human fetal normal colonic mucosa. In Vitro 20:859–868PubMedCrossRefGoogle Scholar
  48. Stein J, Zores M, Schroeder O (2000) Short-chain fatty acid (SCFA) uptake into caco-2 cells by a pH-dependent and carrier mediated transport mechanism. Eur J Nutr 39:121–125PubMedCrossRefGoogle Scholar
  49. Su J, Chen X, Kanekura T (2009) A CD147-targeting siRNA inhibits the proliferation, invasiveness, and VEGF production of human malignant melanoma cells by down-regulating glycolysis. Cancer Lett 273:140–147PubMedCrossRefGoogle Scholar
  50. Thangaraju M, Cresci G, Itagaki S, Mellinger J, Browning DD, Berger FG et al (2008) Sodium-coupled transport of the short chain fatty acid butyrate by SLC5A8 and its relevance to colon cancer. J Gastrointest Surg 12:1773–1781PubMedCrossRefGoogle Scholar
  51. Thibault R, De Coppet P, Daly K, Bourreille A, Cuff M, Bonnet C et al (2007) Down-regulation of the monocarboxylate transporter 1 is involved in butyrate deficiency during intestinal inflammation. Gastroenterology 133:1916–1927PubMedCrossRefGoogle Scholar
  52. Thibault R, Blachier F, Darcy-Vrillon B, de Coppet P, Bourreille A, Segain JP (2010) Butyrate utilization by the colonic mucosa in inflammatory bowel diseases: a transport deficiency. Inflamm Bowel Dis 16:684–695PubMedGoogle Scholar
  53. Tuynman JB, Peppelenbosch MP, Richel DJ (2004) COX-2 inhibition as a tool to treat and prevent colorectal cancer. Crit Rev Oncol Hematol 52:81–101PubMedCrossRefGoogle Scholar
  54. Vaidyanathan JB, Walle T (2003) Cellular uptake and efflux of the tea flavonoid (–)epicatechin-3-gallate in the human intestinal cell line Caco-2. J Pharmacol Exp Ther 307:745–752PubMedCrossRefGoogle Scholar
  55. Wong JMW, Souza R, Kendall CWC, Emam A, Jenkins DJ (2006) Colonic health: fermentation and short chain fatty acids. J Clin Gastroenterol 40:235–243PubMedCrossRefGoogle Scholar
  56. Wood SR, Zhao Q, Smith LH, Daniels CK (2003) Altered morphology in cultured rat intestinal epithelial IEC-6 cells is associated with alkaline phosphatase expression. Tissue Cell 35:47–58PubMedCrossRefGoogle Scholar
  57. Zhang Y, Bao YL, Yang MT, Wu Y, Yu CL, Huang YX et al (2010) Activin A induces SLC5A8 expression through the Smad3 signaling pathway in human colon cancer RKO cells. Int J Biochem Cell Biol 42:1964–1972PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Pedro Gonçalves
    • 1
  • João R. Araújo
    • 1
  • Fátima Martel
    • 1
  1. 1.Department of Biochemistry (U38-FCT), Faculty of MedicineUniversity of PortoPortoPortugal

Personalised recommendations