Share This Article:

Individual and Combined Effects of Nucleotides and Human Milk Oligosaccharides on Proliferation, Apoptosis and Necrosis in a Human Fetal Intestinal Cell Line

Full-Text HTML Download Download as PDF (Size:667KB) PP. 1567-1576
DOI: 10.4236/fns.2012.311205    3,403 Downloads   5,018 Views   Citations


Nucleotides (NT) and human milk oligosaccharides (HMO) individually affect epithelial cell growth, but their combined effects had not been studied. Herein, the impact of NT and HMO on cell proliferation, apoptosis, necrosis and cell cycle in the fetal epithelial cell line (FHs-74 Int) was determined. Cells were incubated with media containing 2.5% FBS and no epidermal growth factor (Control); fucosyllactose (FL) mix [85% 2’FL/15% 3’FL], sialyllactose (SL) mix [40% 6’SL/10% 3’SL/50% sialic acid (SA)] or LNnT at 125, 250, 500 or 1000 μg/mL with and without 250 μg/mL NT (43% CMP, 18.5% UMP, 16.4% AMP, and 22.0% GMP) for 24 or 72 h. NT alone significantly increased proliferation, but did not affect cell cycle or apoptosis/necrosis. All HMO treatments at 1000 μg/mL significantly decreased proliferation and some were also inhibitory at 250 or 500 μg/mL. When NT and HMO were simultaneously added, NT ameliorated the anti-proliferative effect of HMO. FL significantly increased cells in S phase and SL and LNnT treatments significantly increased cells in G2/M and S phases, which concomitantly decreased cells in G0/G1. HMO with NT significantly decreased the percent of cells in the G2/M phase compared to HMO alone. Higher HMO doses significantly increased the percentage of apoptotic and necrotic cells compared to control. In conclusion, HMO reduced cell proliferation and this effect is partially ameliorated by NT. It appears that HMO initially induced apoptosis/necrosis, which was later evidenced by G2/M cell cycle arrest and decreased proliferation.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

S. Hester and S. Donovan, "Individual and Combined Effects of Nucleotides and Human Milk Oligosaccharides on Proliferation, Apoptosis and Necrosis in a Human Fetal Intestinal Cell Line," Food and Nutrition Sciences, Vol. 3 No. 11, 2012, pp. 1567-1576. doi: 10.4236/fns.2012.311205.


[1] J. B. German, C. J. Dillard and R. E. Ward, “Bioactive Components in Milk,” Current Opinion in Clinical Nutrition & Metabolic Care, Vol. 5, No. 6, 2002, pp. 653-658. doi:10.1097/00075197-200211000-00007
[2] E. J. Holen, “Dietary Nucleotides and Intestinal Cell Lines: 1. Modulation of Growth,” Nutrition Research, Vol. 24, 2004, pp. 197-207. doi:10.1016/j.nutres.2003.11.004
[3] S. Kuntz, C. Kunz and S. Rudloff, “Oligosaccharides from Human Milk Induce Growth Arrest via G2/M by Influencing Growth-Related Cell Cycle Genes in Intestinal Epithelial Cells,” British Journal of Nutrition, Vol. 101, No. 9, 2009, pp. 1306-1315. doi:10.1017/S0007114508079622
[4] S. Kuntz, S. Rudloff and C. Kunz, “Oligosaccharides from Human Milk Influence Growth-Related Characteristics of Intestinally Transformed and Non-Transformed Intestinal Cells,” British Journal of Nutrition, Vol. 99, No. 3, 2008, pp. 462-471. doi:10.1017/S0007114507824068
[5] J. L. Leach, J. H. Baxter, B. E. Molitor, et al., “Total Potentially Available Nucleosides of Human Milk by Stage of Lactation,” The American Journal of Clinical Nutrition, Vol. 61, No. 6, 1995, pp. 1224-1230.
[6] J. R. Hess and N. A. Greenberg, “The Role of Nucleotides in the Immune and Gastrointestinal Systems: Potential Clinical Applications,” Nutrition in Clinical Practice, Vol. 27, No. 2, 2012, pp. 281-294. doi:10.1177/0884533611434933
[7] F. Rodriguez-Serrano, J. A. Marchal, A. Rios, et al., “Exogenous Nucleosides Modulate Proliferation of Rat Intestinal Epithelial IEC-6 Cells,” Journal of Nutrition, Vol. 137, No. 4, 2007, pp. 879-884.
[8] D. S. Newburg, “Are All Human Milks Created Equal? Variation in Human Milk Oligosaccharides,” Journal of Pediatric Gastroenterology and Nutrition, Vol. 30, No. 2, 2000, pp. 131-133. doi:10.1097/00005176-200002000-00007
[9] G. V. Coppa, O. Gabrielli, P. Pierani, et al., “Changes in Carbohydrate Composition in Human Milk over 4 Months of Lactation,” Pediatrics, Vol. 9, No. 3, 1993, pp. 637-641.
[10] D. S. Newburg, G. M. Ruiz-Palacios, M. Altaye, et al., “Innate Protection Conferred by Fucosylated Oligosaccharides of Human Milk against Diarrhea in Breastfed Infants,” Glycobiology, Vol. l14, No. 3, 2004, pp. 253-263.
[11] C. Kunz, S. Rudloff, W. Baier, et al., “Oligosaccharides in Human Milk: Structural, Functional, and Metabolic Aspects,” Annual Review of Nutrition, Vol. 20, 2000, pp. 699-722. doi:10.1146/annurev.nutr.20.1.699
[12] L. Bode, “Human Milk Oligosaccharides: Prebiotics and Beyond,” Nutrition Reviews, Vol. 67, Suppl. 2, 2009, pp. S183-S191. doi:10.1111/j.1753-4887.2009.00239.x
[13] S. Asakuma, T. Urashima, M. Akahori, et al., “Variation of Major Neutral Oligosaccharides Levels in Human Colostrum,” European Journal of Clinical Nutrition, Vol. 62, No. 4, 2008, pp. 488-494. doi:10.1038/sj.ejcn.1602738
[14] P. Chaturvedi P, C. D. Warren, M. Altaye, et al., “Fucosylated Human Milk Oligosaccharides Vary between Individuals and over the Course of Lactation,” Glycobiology, Vol. 11, No. 5, 2001, pp. 365-372. doi:10.1093/glycob/11.5.365
[15] M. Musumeci M, J. Simpore, A. D’Agata, et al., “Oligosaccharides in Colostrum of Italian and Burkinabe Women,” Journal of Pediatric Gastroenterology and Nutrition, Vol. 43, No. 3, 2006, pp. 372-378. doi:10.1097/
[16] S. Thurl, M. Munzert, J. Henker, et al., “Variation of Human Milk Oligosaccharides in Relation to Milk Groups and Lactational Periods,” British Journal of Nutrition, Vol. 104, No. 9, 2010, pp. 1-11.
[17] B. Wang, J. Brand-Miller, P. McVeagh, et al., “Concentration and Distribution of Sialic Acid in Human Milk and Infant Formulas,” The American Journal of Clinical Nutrition, Vol. 74, No. 4, 2001, pp. 510-515.
[18] J. B. Miller, S. Bull, J. Miller, et al., “The Oligosaccharide Composition of Human Milk: Temporal and Individual Variations in Monosaccharide Components,” Journal of Pediatric Gastroenterology and Nutrition, Vol. 19, No. 4, 1994. pp. 371-376. doi:10.1097/00005176-199411000-00001
[19] T. Nakano, M. Sugawara and H. Kawakami, “Sialic Acid in Human Milk: Composition and Functions,” Acta paediatrica Taiwanica, Vol. 42, No. 1, 2001, pp. 11-17.
[20] Y. Bao, L. Zhu and D. S. Newburg, “Simultaneous Quantification of Sialyloligosaccharides from Human Milk by Capillary Electrophoresis,” Analytical Biochemistry, Vol. 370, No. 2, 2007, pp. 206-214. doi:10.1016/j.ab.2007.07.004
[21] S. Martin-Sosa, M. J. Martin, L. A. Garcia-Pardo, et al., “Sialyloligosaccharides in Human and Bovine Milk and in Infant Formulas: Variations with the Progression of Lactation,” Journal of Dairy Science, Vol. 86, No. 1, 2003, pp. 52-59. doi:10.3168/jds.S0022-0302(03)73583-8
[22] J. J. Dickson and M. Messer, “Intestinal Neuraminidase Activity of Suckling Rats and Other Mammals. Relationship to the Sialic Acid Content of Milk,” Biochemical Journal, Vol. 170, No. 2, 1978, pp. 407-413.
[23] J. C. Brand-Miller, P. McVeagh, Y. McNeil, et al., “Digestion of Human Milk Oligosaccharides by Healthy Infants Evaluated by the Lactulose Hydrogen Breath Test,” Journal of Pediatrics, Vol. 133, No. 1, 1998. pp. 95-98. doi:10.1016/S0022-3476(98)70185-4
[24] C. S. Potten, “Epithelial Cell Growth and Differentiation. II. Intestinal Apoptosis,” American Journal of Physiology, Vol. 273, No. 2, 1997, pp. G253-257.
[25] S. M. Donovan, “Role of Human Milk Components in Gastrointestinal Development: Current Knowledge and Future Needs,” Journal of Pediatrics, Vol. 149, Suppl. 3, 2006, p. 49.
[26] A. T. Lopez-Navarro, M. A. Ortega, J. Peragon, et al., “Deprivation of Dietary Nucleotides Decreases Protein Synthesis in the Liver and Small Intestine in Rats,” Gastroenterology, Vol. 110, No. 6, 1996, pp. 1760-1769. doi:10.1053/gast.1996.v110.pm8964401
[27] R. Uauy, “Dietary Nucleotides and Requirements in Early Life: Text-Book of Gastroenterology and Nutrition in Infancy,” Raven Press Ltd., New York, 1989.
[28] U. Wenzel, S. Kuntz S, M. D. Brendel, et al., “Dietary Flavone is a Potent Apoptosis Inducer in Human Colon Carcinoma Cells,” Cancer Research, Vol. 60, No. 14, 2000, pp. 3823-3831.
[29] M. Y. Jou, A. F. Philipps, S. L. Kelleher, et al., “Effects of Zinc Exposure on Zinc Transporter Expression in Human Intestinal Cells of Varying Maturity,” Journal of Pediatric Gastroenterology and Nutrition, Vol. 50, No. 6, 2010, pp. 587-595. doi:10.1097/MPG.0b013e3181d98e85
[30] J. C. Chao and S. M. Donovan, “Effects of Insulin, Insulin-Like Growth Factors and Epidermal Growth Factor on Mitogenesis and Disaccharidase Activity in Rat (IEC-6) and Human (FHs 74 Int) Intestinal Cells,” The Chinese Journal of Physiology, Vol. 39, No. 4, 1996, pp. 253-263. doi:10.1146/annurev.nutr.20.1.311
[31] I. R. Sanderson and S. Naik, “Dietary Regulation of Intestinal Gene Expression,” Annual Review of Nutrition, Vol. 20, 2000, pp. 311-338.

comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.