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Functional characterization of three clones of the human intestinal Caco-2 cell line for dietary lipid processing

Published online by Cambridge University Press:  09 March 2007

Séverine Salvini
Affiliation:
Unité 476 INSERM, Nutrition humaine et lipides, 18 Avenue Mozart, 13009 Marseille, France
Monique Charbonnier
Affiliation:
Unité 476 INSERM, Nutrition humaine et lipides, 18 Avenue Mozart, 13009 Marseille, France
Catherine Defoort
Affiliation:
Unité 476 INSERM, Nutrition humaine et lipides, 18 Avenue Mozart, 13009 Marseille, France Laboratoire de Chimie Analytique, Faculté de Pharmacie, 27 Bd J Moulin, 13005 Marseille, France
Christian Alquier
Affiliation:
Unité 476 INSERM, Nutrition humaine et lipides, 18 Avenue Mozart, 13009 Marseille, France
Denis Lairon*
Affiliation:
Unité 476 INSERM, Nutrition humaine et lipides, 18 Avenue Mozart, 13009 Marseille, France
*
*Corresponding author: Dr Denis Lairon, fax +33 491 75 15 62, email lairon@marseille.inserm.fr
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Abstract

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We aimed to improve the use of the human intestinal Caco-2 cell line for studying dietary lipid and cholesterol processing by using isolated pure clones (). Three clones (TC7, PD7 and PF11) were grown as monolayers on semi-permeable filters and compared for cell viability, fatty acid and cholesterol apical uptake or basolateral secretion, apolipoprotein B-48 basolateral secretion and 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase activity. The TC7 clone showed the best viability upon apical incubation with mixed micelles and should be preferred for routine work. Short-term (3·0 h) rates of apical uptake of cholesterol were not different with the three clones, whereas the rate of apical uptake of oleic acid (18 : 1) was lower (P<0·05) with PF11 (250·6 nmol/mg) and the basolateral secretion of cholesterol and oleic acid was lower with the TC7 clone (0·40 and 29·1 nmol/mg respectively). The secretion of apolipoprotein B-48 basolaterally was about 2-fold lower than from PD7 clone. The basal levels of HMG-CoA reductase activity were significantly different (P<0·05; TC7>PF11>PD7). The down-regulation of the enzyme activity was moderate (range 13·8–21·0 %) and comparable in the presence of apical micellar cholesterol, but was much marked upon basolateral incubation with LDL (range 34·0–53·6 %), especially for the PD7 clone. In conclusion, the Caco-2 clones characterized here proved to be particularly suitable for studying lipid nutrients processing. Because these three clones exhibit some different metabolic capabilities, they provide a new tool to study intestinal response to lipid nutrients.

Type
Research Article
Copyright
Copyright © The Nutrition Society 2002

References

Anderberg, EK & Artursson, P (1993) Epithelial transport of drugs in cell culture. VIII: Effects of sodium dodecyl sulfate on cell membrane and tight junction permeability in human intestinal epithelial (Caco-2) cells. Journal of Pharmaceutical Sciences 82, 392398.CrossRefGoogle ScholarPubMed
Beaumier-Gallon, G, Dubois, C, Senft, M, Vergnes, MF, Pauli, AM, Portugal, H & Lairon, D (2001) Dietary cholesterol is secreted in intestinally derived chylomicrons during several subsequent postprandial phases in healthy humans. American Journal of Clinical Nutrition 73, 870877.CrossRefGoogle ScholarPubMed
Bosner, MS, Lange, LG, Stenson, WF & Ostlund, RE Jr (1999) Percent cholesterol absorption in normal women and men quantified with dual stable isotopic tracers and negative ion mass spectrometry. Journal of Lipid Research 40, 302308.CrossRefGoogle ScholarPubMed
Brown, MS, Dana, SE & Goldstein, JL (1973) Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in human fibroblasts by lipoproteins. Proceedings of the National Academy Sciences, USA 70, 21622166.CrossRefGoogle Scholar
Brown, MS & Goldstein, JL (1980) Multivalent feedback regulation of HMG CoA reductase, a control mechanism coordinating isoprenoid synthesis and cell growth. Journal of Lipid Research 21, 505517.CrossRefGoogle ScholarPubMed
Caro, I, Boulenc, X, Rousset, M, Meunier, V, Bourrié, M, Julian, B, Joyeux, H, Roques, C, Berger, Y, Zweibaum, A & Fabre, G (1995) Characterisation of a newly isolated Caco-2 clone (TC7), as a model of transport processes and biotransformation of drugs. International Journal of Pharmaceutics 116, 147158.CrossRefGoogle Scholar
Chantret, I, Rodolosse, A, Barbat, A, Dussaulx, E, Brot-Laroche, E, Zweibaum, A & Rousset, M (1994) Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: evidence for glucose-dependent negative regulation. Journal of Cell Science 107, 213225.CrossRefGoogle ScholarPubMed
Chong, MW, Gu, KD, Lam, PK, Yang, M & Fong, WF (2000) Study on the cytotoxicity of microcystin-LR on cultured cells. Chemosphere 41, 143147.CrossRefGoogle Scholar
Dietschy, JM & Gamel, WG (1971) Cholesterol synthesis in the intestine of man: regional differences and control mechanisms. Journal of Clinical Investigation 50, 872880.CrossRefGoogle ScholarPubMed
Dubois, C, Beaumier, G, Juhel, C, Armand, M, Portugal, H, Pauli, AM, Borel, P, Latge, C & Lairon, D (1998) Effects of graded amounts (0–50g) of dietary fat on postprandial lipemia and lipoproteins in normolipidemic adults. American Journal of Clinical Nutrition 67, 3138.CrossRefGoogle Scholar
Field, FJ, Born, E & Mathur, SN (1997) Effect of micellar beta-sitosterol on cholesterol metabolism in CaCo-2 cells. Journal of Lipid Research 38, 348360.CrossRefGoogle ScholarPubMed
Field, FJ, Fujiwara, D, Born, E, Chappell, DA & Mathur, SN (1993) Regulation of LDL receptor expression by luminal sterol flux in Caco-2 cells. Thrombosis 13, 729737.Google ScholarPubMed
Field, FJ, Shreves, T, Fujiwara, D, Murthy, S, Albright, E & Mathur, SN (1991) Regulation of gene expression and synthesis and degradation of 3-hydroxy-3-methylglutaryl coenzyme A reductase by micellar cholesterolin CaCo-2 cells. Journal of Lipid Research 32, 18111821.CrossRefGoogle Scholar
Halleux, C & Schneider, YJ (1991) Iron absorption by intestinal epithelial cells: 1. CaCo2 cells cultivated in serum-free medium, on polyethyleneterephthalate microporous membranes, as an in vitro model. In Vitro Cellular and Developmental Biology 27A, 293302.CrossRefGoogle ScholarPubMed
Hauser, H, Dyer, JH, Nandy, A, Vega, MA, Werder, M, Bieliauskaite, E, Weber, FE, Compassi, S, Gemperli, A, Boffelli, D, Wehrli, E, Schulthess, G & Phillips, MC (1998) Identification of a receptor mediating absorption of dietary cholesterol in the intestine. Biochemistry 37, 1784317850.CrossRefGoogle ScholarPubMed
Homan, R & Hamelehle, KL (1998) Phospholipase A2 relieves phosphatidylcholine inhibition of micellar cholesterol absorption and transport by human intestinal cell line Caco-2. Journal of Lipid Research 39, 11971209.CrossRefGoogle ScholarPubMed
Hussain, MM (2000) A proposed model for the assembly of chylomicrons. Atherosclerosis 148, 115.CrossRefGoogle ScholarPubMed
Levy, E, Mehran, M & Seidman, E (1995) Caco-2 cells as a model for intestinal lipoprotein synthesis and secretion. FASEB Journal 9, 626635.CrossRefGoogle Scholar
Lorec, AM, Juhel, C, Pafumi, Y, Portugal, H, Pauli, AM, Lairon, D & Defoort, C (2000) Determination of apolipoprotein B-48 in plasma by a competitive ELISA. Clinical Chemistry 46, 16381642.CrossRefGoogle ScholarPubMed
Mahraoui, L, Takeda, J, Mesonero, J, Chantret, I, Dussaulx, E, Bell, GI & Brot-Laroche, E (1994) Regulation of expression of the human fructose transporter (GLUT5) by cyclic AMP. Biochemical Journal 301, 169175.CrossRefGoogle ScholarPubMed
Mamo, JC, Proctor, SD & Smith, D (1998) Retention of chylomicron remnants by arterial tissue; importance of an efficient clearance mechanism from plasma. Atherosclerosis 141 Suppl. 1, S63S69.CrossRefGoogle ScholarPubMed
Mehran, M, Levy, E, Bendayan, M & Seidman, E (1997) Lipid, apolipoprotein, and lipoprotein synthesis and secretion during cellular differentiation in Caco-2 cells. In Vitro Cellular and Developmental Biology 33, 118128.CrossRefGoogle ScholarPubMed
Pinto, M, Robine-Leon, S, Appay, MD, Kedinger, M, Triadou, N, Dussaulx, E, Lacroix, B, Simon-Assmann, P, Haffen, K, Fogh, J & Zweibaum, A (1983) Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biology of the Cell 47, 323330.Google Scholar
Reimann, FM, Herold, G, Grosshans, I, Rogler, G, Fellermann, K & Stange, EF (1992) Regulation of cholesterol metabolism and low-density lipoprotein binding in human intestinal Caco-2 cells. Digestion 51, 1017.CrossRefGoogle ScholarPubMed
Sehayek, E, Ono, JG, Shefer, S, Nguyen, LB, Wang, N, Batta, AK, Salen, G, Smith, JD, Tall, AR & Breslow, JL (1998) Biliary cholesterol excretion: a novel mechanism that regulates dietary cholesterol absorption. Proceedings of the National Academy Sciences, USA 95, 1019410199.CrossRefGoogle ScholarPubMed
Tranchant, T, Besson, P, Hoinard, C, Delarue, J, Antoine, JM, Couet, C & Gore, J (1997) Mechanisms and kinetics of alpha-linolenic acid uptake in Caco-2 clone TC7. Biochimica et Biophysica Acta 1345, 151161.CrossRefGoogle ScholarPubMed
Trotter, PJ, Ho, SY & Storch, J (1996) Fatty acid uptake by Caco-2 human intestinal cells. Journal of Lipid Research 37, 336346.CrossRefGoogle ScholarPubMed
van Greevenbroek, MM, Erkelens, DW & de Bruin, TW (2000) Caco-2 cells secrete two independent classes of lipoproteins with distinct density: effect of the ratio of unsaturated to saturated fatty acid. Atherosclerosis 149, 2531.CrossRefGoogle ScholarPubMed
van Greevenbroek, MM, Van Meer, G, Erkelens, DW & de Bruin, TW (1996) Effects of saturated, mono-, and polyunsaturated fatty acids on the secretion of apo B containing lipoproteins by Caco-2 cells. Atherosclerosis 121, 139150.CrossRefGoogle ScholarPubMed