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The effect of oestrogen and dietary phyto-oestrogens on transepithelial calcium transport in human intestinal-like Caco-2 cells

Published online by Cambridge University Press:  09 March 2007

Alice A. Cotter
Affiliation:
Department of Food and Nutritional Sciences, University College, Cork, Ireland
Christopher Jewell
Affiliation:
Department of Food and Nutritional Sciences, University College, Cork, Ireland
Kevin D. Cashman*
Affiliation:
Department of Food and Nutritional Sciences, University College, Cork, Ireland Department of Medicine, University College, Cork, Ireland
*
*Corresponding author: Professor Kevin D. Cashman, fax +353 21 4270244, email k.cashman@ucc.ie
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Abstract

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Recently, dietary phyto-oestrogens (PO) have been suggested as possible alternatives to oestrogen therapy (hormone replacement therapy) as a means of preventing bone loss associated with ovarian hormone deficiency. While PO, which exhibit oestrogen-like activity, act directly on bone cells, their protective effect on bone may be partly due to their ability to enhance Ca absorption. Therefore, the aim of the present study was to investigate the effect of 17β-oestradiol and two commonly consumed soyabean PO (genistein and daidzein) on Ca absorption in the human Caco-2 intestinal-like cell model. Caco-2 cells were seeded onto permeable filter supports and allowed to differentiate into monolayers. On day 21, the Caco-2 monolayers (n 8–18 per treatment), grown in oestrogen-replete or -deplete media, were then exposed to 10 nM-17β-oestradiol, 1 nM-1,25-dihydroxycholecalciferol, or 50 μM-genistein or -daidzein for 24 h. After exposure, transepithelial and transcellular transport of 45Ca and fluorescein transport (a marker of paracellular diffusion) were measured. As expected, 1,25-dihydroxycholecalciferol stimulated Ca absorption in Caco-2 cells, by up-regulating transcellular transport, whereas 17β-oestradiol had no effect on Ca absorption. Unexpectedly, both PO decreased Ca absorption (by about 17–19 % compared with control, P<0·05), by reducing transcellular Ca transport in Caco-2 cells grown in oestrogen-replete media. This inhibitory effect disappeared when monolayers were grown in oestrogen-deplete media. In conclusion, PO at high luminal concentrations either had no effect or reduced Ca absorption in Caco-2 cells, dependent on oestrogen status. The effect of lower concentrations of these compounds needs to be investigated.

Type
Research Article
Copyright
Copyright © The Nutrition Society 2003

References

Agnusdei, D, Crepaldi, G, Isaia, G, Mazzuoli, G, Ortolani, S, Passeri, M, Bufalino, L & Gennari, C (1997) A double blind, placebo-controlled trial of ipriflavone for prevention of postmenopausal spinal bone loss. Calcified Tissue International 61, 142147.CrossRefGoogle ScholarPubMed
Akiyama, T, Ishida, J & Nakagawa, S (1987) Genistein: a specific inhibitor of tyrosine-specific protein kinase. Journal of Biological Chemistry 262, 55925595.CrossRefGoogle Scholar
Alekel, DL, Germain, AS, Peterson, CT, Hanson, KB, Stewart, JW & Toda, T (2000) Isoflavone-rich soy protein isolate attenuates bone loss in the lumbar spine of perimenopausal women. American Journal of Clinical Nutrition 72, 844852.CrossRefGoogle ScholarPubMed
Anderson, JJ, Ambrose, WW & Garner, SC (1998) Biphasic effects of genistein on bone tissue in the ovariectomised, lactating rat model. Proceedings of the Society for Experimental Biology and Medicine 217, 345350.CrossRefGoogle Scholar
Anderson, JJB, Anthony, M, Messina, M & Garner, SC (1999) Effects of phytoestrogens in tissues. Nutrition Research Reviews 12, 75116.CrossRefGoogle Scholar
Arai, N, Strom, A, Rafter, JJ & Gustafsson, JA (2000) Estrogen receptor beta mRNA in colon cancer cells: growth effects of estrogen and genistein. Biochemical and Biophysical Research Communications 13, 425431.CrossRefGoogle Scholar
Arjmandi, BH & Smith, BJ (2002) Soy isoflavones’ osteoprotective role in postmenopausal women: mechanism of action. Journal of Nutritional Biochemistry 13, 130137.CrossRefGoogle ScholarPubMed
Arjmandi, BH (2001) The role of phytoestrogens in the prevention and treatment of osteoporosis in ovarian hormone deficiency. Journal of American College Nutrition 20, 398S402S.CrossRefGoogle ScholarPubMed
Arjmandi, BH, Alekel, L, Hollis, BW, Amin, D, Stacewicz-Sapuntzakis, M, Guo, P & Kukreja, SC (1996) Dietary soybean protein prevents bone loss in an ovariectomized rat model of osteoporosis. Journal of Nutrition 126, 161167.CrossRefGoogle Scholar
Arjmandi, BH, Birnbaum, R, Goyal, NV, Getlinger, MJ, Juma, S, Alekel, L, Hasler, CM, Drum, ML, Hollis, BW & Kukreja, SC (1998 a) Bone-sparing effect of soy protein in ovarian hormone-deficient rats is related to its isoflavone content. American Journal of Clinical Nutrition 68, 1346S1368S.Google ScholarPubMed
Arjmandi, BH, Getlinger, MJ, Goyal, NV, Alekel, L, Hasler, CM, Juma, S, Drum, ML, Hollis, BW & Kukreja, SC (1998 b) Role of soy protein with normal or reduced isoflavone content in reversing bone loss induced ovarian hormone deficiency in rats. American Journal of Clinical Nutrition 68, 1358S1363S.CrossRefGoogle ScholarPubMed
Arjmandi, BH, Khalil, DA & Hollis, BW (2000) Ipriflavone, a synthetic phytoestrogen, enhances intestinal calcium transport in vitro. Calcified Tissue International 67, 225229.CrossRefGoogle ScholarPubMed
Arjmandi, BH, Khalil, DA & Hollis, BW (2002) Soy protein: its effect on intestinal calcium transport, serum vitamin D, and insulin-like growth factor-1 in various ovariectomized rats. Calcified Tissue International 70, 483487.CrossRefGoogle ScholarPubMed
Arjmandi, BH, Salih, MA, Herbert, DC, Sims, SH & Kalu, DN (1993) Evidence for estrogen receptor-linked calcium transport in the intestine. Bone and Mineral 21, 6374.CrossRefGoogle ScholarPubMed
Blair, HC, Jordan, SE, Peterson, TG & Barnes, S (1996) Variable effects of tyrosine kinase inhibitors on avian osteoclastic activity and reduction of bone loss in ovariectomised rats. Journal of Cell Biochemistry 61, 629637.3.0.CO;2-A>CrossRefGoogle Scholar
Bronner, F, Pansu, D & Stein, WD (1986) An analysis of intestinal calcium transport across the rat intestine. American Journal of Physiology 250, G561G569.Google ScholarPubMed
Campbell-Thompson, M, Lynch, IJ & Bhardwaj, B (2001) Expression of estrogen receptor (ER) subtypes and ERbeta isoforms in colon cancer. Cancer Research 61, 632640.Google Scholar
Capiati, D, Benassati, S & Boland, RL (2002) 1,25(OH)2-vitamin D3 induces translocation of the vitamin D receptor (VDR) to the plasma membrane in skeletal muscle cells. Journal of Cell Biochemistry 86, 128135.CrossRefGoogle Scholar
Cassidy, A (1996) Physiological effects of phyto-oestrogens in relation to cancer and other human health risks. Proceedings of the Nutrition Society 55, 399417.CrossRefGoogle ScholarPubMed
Chirayath, MV, Gajdzik, L, Hulla, W, Graf, J, Cross, HS & Peterlik, M (1998) Vitamin D increases tight-junction conductance and paracellular Ca2+ transport in Caco-2 cell cultures. American Journal of Physiology 274, G389G396.Google ScholarPubMed
Colin, EM, Van Den Bemd, GJ, Van Aken, M, Christakos, S, De Jonge, HR, Deluca, HF, Prahl, JM, Birkenhager, JC, Buurman, CJ, Pols, HA & Van Leeuwen, JP (1999) Evidence for involvement of 17beta-estradiol in intestinal calcium absorption independent of 1,25-dihydroxyvitamin D3 level in the rat. Journal of Bone and Mineral Research 14, 5764.CrossRefGoogle ScholarPubMed
Dalais, FS, Rice, GE, Wahlquist, ML, Grehan, M, Murkies, AL, Medley, G, Ayton, R & Strauss, BJG (1998) Effects of dietary phytoestrogens in postmenopausal women. Climacteric 1, 124129.CrossRefGoogle ScholarPubMed
Di Domenico, M, Castoria, G, Bilancio, A, Migliaccio, A & Auricchio, F (1996) Estradiol activation of human colon carcinoma-derived Caco-2 cell growth. Cancer Research 56, 45164521.Google ScholarPubMed
Edmonson, JM, Armstrong, LS & Martinez, AO (1988) A rapid and simple MTT-based assay for determining drug sensitivity in monolayer cultures. Journal of Tissue Culture Methods 11, 1517.CrossRefGoogle Scholar
Fleet, JC & Wood, RJ (1994) Identification of calbindin D-9k mRNA and its regulation by 1,25-dihydroxyvitamin D3 in Caco-2 cells. Archives of Biochemistry and Biophysics 308, 171174.CrossRefGoogle ScholarPubMed
Fleet, JC & Wood, RJ (1999) Specific 1,25(OH)2D3-mediated regulation of transcellular calcium transport in Caco-2 cells. American Journal of Physiology 276, G958G964.Google Scholar
Fleet, JC, Bradley, J, Reddy, GS, Ray, R & Wood, RJ (1996) 1 alpha,25-(OH)2-vitamin D3 analogs with minimal in vivo calcemic activity can stimulate significant transepithelial calcium transport and mRNA expression in vitro. Archives of Biochemistry and Biophysics 15, 228234.CrossRefGoogle Scholar
Gallagher, JC (1990) The pathogenesis of osteoporosis. Bone Mineral 9, 215227.CrossRefGoogle ScholarPubMed
Gallagher, JC (2001) Role of estrogens in the management of postmenopausal bone loss. Rheumatic Diseases Clinics of North America 27, 143162.CrossRefGoogle ScholarPubMed
Gallagher, JC, Riggs, BL & DeLuca, HF (1980) Effect of estrogen on calcium absorption and serum vitamin D metabolites in postmenopausal osteoporosis. Journal of Clinical Endocrinology and Metabolism 51, 13591364.Google Scholar
Gao, YH & Yamaguchi, M (2000) Suppressive effect of genistein on rat bone osteoclasts: Involvement of protein kinase inhibition and protein tyrosine phosphatase activation. International Journal of Molecular Medicine 5, 261267.Google ScholarPubMed
Gennari, C, Agnusdei, D, Crepaldi, G, Isaia, G, Mazzuoli, G, Ortolani, S, Bufalino, L & Passeri, M (1998) Effect of ipriflavone – a synthetic derivative of natural isoflavones – on bone mass loss in the early years after menopause. Menopause 5, 915.Google Scholar
Giuliano, AR, Franceschi, RT & Wood, RJ (1991) Characterization of the vitamin D receptor from the Caco-2 human colon carcinoma cell line: effect of cellular differentiation. Archives of Biochemistry and Biophysics 285, 261269.CrossRefGoogle ScholarPubMed
Giuliano, AR & Wood, RJ (1991) Vitamin D-regulated calcium transport in Caco-2 cells: unique in vitro model. American Journal of Physiology 260, G207G212.Google ScholarPubMed
Heaney, RP, Recker, RR & Saville, PD (1978) Menopausal changes in calcium balance performance. Journal of Laboratory and Clinical Medicine 92, 953963.Google Scholar
Hsu, CS, Shen, WW, Hsueh, YM & Yeh, SL (2001) Soy isoflavone supplementation in postmenopausal women. Effects on plasma lipids, antioxidant enzyme activities and bone density. Journal of Reproductive Medicine 46, 221226.Google Scholar
Hunt, SM, Chrzanowski, C, Barnett, CR, Brand, HN & Fawell, JK (1987) A comparison of in vitro cytotoxicity assays and their application to water samples. Alternatives to Laboratory Animals 15, 2029.Google Scholar
Klinge, CM (2001) Estrogen receptor interaction with estrogen response elements. Nucleic Acids Research 29, 29052919.CrossRefGoogle ScholarPubMed
Kuiper, GG, Carlsson, B, Grandien, K, Enmark, E, Haggblad, J, Nilsson, S & Gustafsson, JA (1997) Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology 138, 863870.CrossRefGoogle ScholarPubMed
Lephart, ED, Thompson, JM, Setchell, KD, Adlercreutz, H & Weber, KS (2000) Phytoestrogens decrease brain calcium-binding proteins but do not alter hypothalmic androgen metabolizing enzymes in adult male rat. Brain Research 859, 123131.CrossRefGoogle ScholarPubMed
Lindmark, T, Kimura, Y & Artursson, P (1998) Absorption enhancement through intracellular regulation of tight junction permeability by medium chain fatty acids in Caco-2 cells. Journal of Pharmacology and Experimental Therapeutics 284, 362369.Google ScholarPubMed
Markovits, J, Linassier, C, Fosse, P, Couprie, J, Pierre, J, Jacquemin-Sablon, A, Saucier, JM, Le Pecq, JB & Larsen, AK (1989) Inhibitory effects of the tyrosine kinase inhibitor genistein on mammalian DNA topoisomerase II. Cancer Research 49, 51115117.Google ScholarPubMed
Messina, MJ, Persky, V, Setchell, KD & Barnes, S (1994) Soy intake and cancer risk: a review of the in vitro and in vivo data. Nutrition and Cancer 21, 113131.Google Scholar
Miksicek, RJ (1995) Estrogenic flavonoids: structural requirements for biological activity. Proceedings of the Society for Experimental Biology and Medicine 208, 4450.CrossRefGoogle ScholarPubMed
Morabito, N, Crisafulli, A, Vergara, C, Gaudio, A, Lasco, A, Frisina, N, D'Anna, R, Corrado, F, Pizzoleo, MA, Cincotta, M, Altavilla, D, Ientile, R & Squadrito, F (2002) Effects of genistein and hormone-replacement therapy on bone loss in early postmenopausal women: a randomized double-blind placebo-controlled study. Journal of Bone and Mineral Research 17, 19041912.Google Scholar
Mossman, T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65, 5563.CrossRefGoogle Scholar
Nguyen, TV, Jones, G, Sambrook, PN, White, CP, Kelly, PJ & Eisman, JA (1995) Effects of estrogen exposure and reproductive factors on bone mineral density and osteoporotic fractures. Journal of Clinical Endocrinology and Metabolism 80, 27092714.Google Scholar
Norman, AW, Okamura, WH, Bishop, JE & Henry, HL (2002) Update on biological actions of 1α,25(OH)2-vitamin D3 (rapid effects) and 24R,25(OH)2-vitamin D3. Molecular and Cellular Endocrinology 197, 113.CrossRefGoogle Scholar
Ohta, H, Komukai, S, Makita, K, Masuzawa, T & Nozawa, S (1999) Effects of 1-year ipriflavone treatment on lumbar bone mineral density and bone metabolic markers in postmenopausal women with low bone mass. Hormone Research 51, 178183.CrossRefGoogle ScholarPubMed
O'Loughlin, PD & Morris, HA (1998) Oestrogen deficiency impairs intestinal calcium absorption in the rat. Journal of Physiology 15, 313322.CrossRefGoogle Scholar
Omi, N, Aoi, S, Murata, K & Ezawa, I (1992) The effect of soybean milk on bone mineral density and bone strength in ovariectomized osteoporotic rats. Journal of Home Economics in Japan 44, 549554.Google Scholar
Omi, N, Aoi, S, Murata, K & Ezawa, I (1994) Evaluation of the effect of soybean milk and soybean milk peptide on bone metabolism in the rat model with ovariectomised osteoporosis. Journal of Nutritional Science and Vitaminology 40, 201211.CrossRefGoogle Scholar
Picherit, C, Chanteranne, B, Bennetau-Pelissero, C, Davicco, M-J, Lebecque, P, Bartlett, J-P & Coxam, V (2001) Dose-dependent bone-sparing effects on dietary isoflavones in the ovariectomised rat. British Journal of Nutrition 85, 307316.CrossRefGoogle ScholarPubMed
Picherit, C, Coxam, V, Bennetau-Pelissero, C, Kati-Coulibaly, S, Davicco, MJ, Lebecque, P & Bartlett, JP (2000) Daidzein is more efficient than genistein in preventing ovariectomy-induced bone loss in rats. Journal of Nutrition 130, 16751681.Google ScholarPubMed
Pinto, M, Robine-Leon, S, Appay, MD, Kedinger, M, Triadow, N, Dussaulx, E, Lacroix, B, Simon-Assmann, P, Haffer, 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
Potter, SM, Baum, JA, Teng, H, Stillman, RJ, Shay, NF & Erdman, JW Jr (1998) Soy protein and isoflavones: their effects on blood lipids and bone density in postmenopausal women. American Journal of Clinical Nutrition 68, 1375S1379S.CrossRefGoogle ScholarPubMed
Riggs, BL & Melton, LJ 3rd (1986) Involutional osteoporosis. New England Journal of Medicine 26, 16761686.Google Scholar
Shao, A, Wood, RJ & Fleet, JC (2001) Increased vitamin D receptor level enhances 1,25-dihydroxyvitamin D3-mediated gene expression and calcium transport in Caco-2 cells. Journal of Bone and Mineral Research 16, 615624.CrossRefGoogle Scholar
Snedecor, GW & Cochran, WG (1967) Statistical Methods. Ames, IA: Iowa State University Press.Google Scholar
Taylor, H, Quintero, EM, Iacopino, AM & Lephart, ED (1999) Phytoestrogens alter hypothalamic calbindin-D28k levels during prenatal development. Brain Research and Developmental Brain Research 14, 277281.CrossRefGoogle Scholar
Taylor, M (1997) Alternatives to conventional hormone replacement therapy. Comprehensive Therapy 23, 514532.Google ScholarPubMed
Ten Bolscher, M, Netelenbos, JC, Barto, R, Van Buuren, LM & Van der vijgh, WJ (1999) Estrogen regulation of intestinal calcium absorption in the intact and ovariectomized adult rat. Journal of Bone and Mineral Research 14, 11971202.CrossRefGoogle ScholarPubMed
Vassault, A (1983) In Methods of Enzymatic Analysis, 3rd ed., vol. 3, pp. 118126 [Bergmeyer, HV, editor]. Deerfield Beach, FL: VCH Weinheim, West Germany.Google Scholar
Welshons, WV, Wolf, MF, Murphy, CS & Jordan, VC (1988) Estrogenic activity of Phenol Red. Molecular and Cellular Endocrinology 57, 169178.Google Scholar
Wood, RJ, Tchack, L & Taparia, S (2001) 1,25-Dihydroxyvitamin D3 increases the expression of the CaT1 epithelial calcium channel in the Caco-2 human intestinal cell line. BMC Physiology 1, 11.CrossRefGoogle ScholarPubMed
Yamaguchi, M & Sugimoto, E (2000) Stimulatory effect of genistein and diadzein on protein synthesis in osteoblastic MC3T3-E1 cells: Activation of aminoacyl-tRNA synthetase. Molecular and Cellular Biochemistry 214, 97102.CrossRefGoogle ScholarPubMed
Yee, S (1997) In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man – fact or myth. Pharmacological Research 14, 763766.CrossRefGoogle ScholarPubMed
Zava, DT & Duwe, G (1997) Estrogenic and antiproliferative properties of genistein and other flavonoids in human breast cancer cells in vitro. Nutrition and Cancer 27, 3140.Google Scholar