DOI QR코드

DOI QR Code

Mammary alveolar cell as in vitro evaluation system for casein gene expression involved in glucose level

  • Heo, Young Tae (Department of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University) ;
  • Ha, Woo Tae (Department of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University) ;
  • Lee, Ran (Department of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University) ;
  • Lee, Won-Young (Division of Food Bioscience, RIBHS, College of Biomedical and Health Sciences, Konkuk University) ;
  • Jeong, Ha Yeon (Department of Animal Resources Development Dairy Science Division, National Institute of Animal Science, RDA) ;
  • Hwang, Kyu Chan (Sooam Biotech Research Foundations) ;
  • Song, Hyuk (Department of Stem Cell and Regenerative Biology, College of Animal Bioscience and Technology, Konkuk University)
  • Received : 2016.07.05
  • Accepted : 2016.09.19
  • Published : 2017.06.01

Abstract

Objective: Glucose is an essential fuel in the energy metabolism and synthesis pathways of all mammalian cells. In lactating animals, glucose is the major precursor for lactose and is a substrate for the synthesis of milk proteins and fat in mammary secretory (alveolar) epithelial cells. However, clear utilization of glucose in mammary cells during lactogenesis is still unknown, due to the lack of in vitro analyzing models. Therefore, the objective of this study was to test the reliability of the mammary alveolar (MAC-T) cell as an in vitro study model for glucose metabolism and lactating system. Methods: Undifferentiated MAC-T cells were cultured in three types of Dulbecco's modified Eagle's medium with varying levels of glucose (no-glucose: 0 g/L, low-glucose: 1 g/L, and high-glucose: 4.5 g/L) for 8 d, after which differentiation to casein secretion was induced. Cell proliferation and expression levels of apoptotic genes, Insulin like growth factor-1 (IGF1) receptor, oxytocin receptor, ${\alpha}S1$, ${\alpha}S2$, and ${\beta}$ casein genes were analyzed at 1, 2, 4, and 8 d after differentiation. Results: The proliferation of MAC-T cells with high-glucose treatment was seen to be significantly higher. Expression of apoptotic genes was not affected in any group. However, expression levels of the mammary development related gene (IGF1 receptor) and lactation related gene (oxytocin receptor) were significantly higher in the low-glucose group. Expressions of ${\alpha}S1-casein$, ${\alpha}S2-casein$, and ${\beta}-casein$ were also higher in the low-glucose treated group as compared to that in the no-glucose and high-glucose groups. Conclusion: The results demonstrated that although a high-glucose environment increases cell proliferation in MAC-T cells, a low-glucose treatment to MAC-T cells induces higher expression of casein genes. Our results suggest that the MAC-T cells may be used as an in vitro model to analyze mammary cell development and lactation connected with precise biological effects.

Keywords

References

  1. Cardenas H, Carvajal A, Utreras E, et al. Lactation inhibits the potentiating effect of galanin upon the GnRH-induced LH release observed in diestrous-1 rat. Biol Res 1998;31:351-8.
  2. Threadgold LC, Kuhn NJ. Glucose-6-phosphate hydrolysis by lactating rat mammary gland. Int J Biochem 1979;10:683-5. https://doi.org/10.1016/0020-711X(79)90212-X
  3. Kuhn NJ, Carrick DT, Wilde CJ. Lactose synthesis: the possibilities of regulation. J Dairy Sci 1980;63:328-36. https://doi.org/10.3168/jds.S0022-0302(80)82934-1
  4. Stacey A, Schnieke A, Kerr M, et al. Lactation is disrupted by alphalactalbumin deficiency and can be restored by human alpha-lactalbumin gene replacement in mice. Proc Natl Acad Sci USA 1995;92: 2835-9. https://doi.org/10.1073/pnas.92.7.2835
  5. Xiao CT, Cant JP. Relationship between glucose transport and metabolism in isolated bovine mammary epithelial cells. J Dairy Sci 2005; 88:2794-805. https://doi.org/10.3168/jds.S0022-0302(05)72959-3
  6. Rulquin H, Rigout S, Lemosquet S, Bach A. Infusion of glucose directs circulating amino acids to the mammary gland in well-fed dairy cows. J Dairy Sci 2004;87:340-9. https://doi.org/10.3168/jds.S0022-0302(04)73173-2
  7. Brown EG, Vandehaar MJ, Daniels KM, et al. Effect of increasing energy and protein intake on mammary development in heifer calves. J Dairy Sci 2005;88:595-603. https://doi.org/10.3168/jds.S0022-0302(05)72723-5
  8. Al-Trad B, Reisberg K, Wittek T, et al. Increasing intravenous infusions of glucose improve body condition but not lactation performance in midlactation dairy cows. J Dairy Sci 2009;92:5645-58. https://doi.org/10.3168/jds.2009-2264
  9. Blum JW, Schnyder W, Kunz PL, et al. Reduced and compensatory growth: endocrine and metabolic changes during food restriction and refeeding in steers. J Nutr 1985;115:417-24. https://doi.org/10.1093/jn/115.4.417
  10. Daniels KM, McGilliard ML, Meyer MJ, et al. Effects of body weight and nutrition on histological mammary development in Holstein heifers. J Dairy Sci 2009;92:499-505. https://doi.org/10.3168/jds.2008-1007
  11. Zavizion B, van Duffelen M, Schaeffer W, Politis I. Establishment and characterization of a bovine mammary epithelial cell line with unique properties. In Vitro Cell Dev Biol Anim 1996;32:138-48. https://doi.org/10.1007/BF02723679
  12. Huynh HT, Robitaille G, Turner JD. Establishment of bovine mammary epithelial cells (MAC-T): an in vitro model for bovine lactation. Exp Cell Res 1991;197:191-9. https://doi.org/10.1016/0014-4827(91)90422-Q
  13. Thorn SR, Purup S, Vestergaard M, et al. Regulation of mammary parenchymal growth by the fat pad in prepubertal dairy heifers: role of inflammation-related proteins. J Endocrinol 2008;196:539-46. https://doi.org/10.1677/JOE-07-0501
  14. Purup S, Nielsen TS. Cell-based models to test the effects of milkderived bioactives. Animal 2012;6:423-32. https://doi.org/10.1017/S1751731111002540
  15. Cohick WS, Turner JD. Regulation of IGF binding protein synthesis by a bovine mammary epithelial cell line. J Endocrinol 1998;157: 327-36. https://doi.org/10.1677/joe.0.1570327
  16. Berry SDK, Nielsen MSW, Sejrsen K, et al. Use of an immortalized bovine mammary epithelial cell line (MAC-T) to measure the mitogenic activity of extracts from heifer mammary tissue: effects of nutrition and ovariectomy. Domest Anim Endocrinol 2003;25: 245-53. https://doi.org/10.1016/S0739-7240(03)00062-6
  17. Peterson DG, Matitashvili EA, Bauman DE. The inhibitory effect of trans-10, cis-12 CLA on lipid synthesis in bovine mammary epithelial cells involves reduced proteolytic activation of the transcription factor SREBP-1. J Nutr 2004;134:2523-7. https://doi.org/10.1093/jn/134.10.2523
  18. Thorn SR, Purup S, Cohick WS, et al. Leptin does not act directly on mammary epithelial cells in prepubertal dairy heifers. J Dairy Sci 2006;89:1467-77. https://doi.org/10.3168/jds.S0022-0302(06)72214-7
  19. Bruzelius K, Purup S, James P, Onning G, Akesson B. Biosynthesis of selenoproteins in cultured bovine mammary cells. J Trace Elem Med Biol 2008;22:224-33. https://doi.org/10.1016/j.jtemb.2008.03.009
  20. Sorensen BM, Chris Kazala E, Murdoch GK, et al. Effect of CLA and other C18 unsaturated fatty acids on DGAT in bovine milk fat biosynthetic systems. Lipids 2008;43:903-12. https://doi.org/10.1007/s11745-008-3216-z
  21. Zhou Y, Capuco AV, Jiang H. Involvement of connective tissue growth factor (CTGF) in insulin-like growth factor-I (IGF1) stimulation of proliferation of a bovine mammary epithelial cell line. Domest Anim Endocrinol 2008;35:180-9. https://doi.org/10.1016/j.domaniend.2008.05.003
  22. Naso LG, Lezama L, Rojo T, et al. Biological evaluation of morin and its new oxovanadium(IV) complex as antio-xidant and specific anti-cancer agents. Chem Biol Interact 2013;206:289-301. https://doi.org/10.1016/j.cbi.2013.10.006
  23. Wang B, Zhao M-z, Cui N-p, et al. Kruppel-like factor 4 induces apoptosis and inhibits tumorigenic progression in SK-BR-3 breast cancer cells. FEBS Open Bio 2015;5:147-54. https://doi.org/10.1016/j.fob.2015.02.003
  24. Lee HY, Heo YT, Lee SE, et al. Short communication: retinoic acid plus prolactin to synergistically increase specific casein gene expression in MAC-T cells. J Dairy Sci 2013;96:3835-9. https://doi.org/10.3168/jds.2012-5945
  25. Lyons WR. Hormonal synergism in mammary growth. Proc R Soc Lond B Biol Sci 1958;149:303-25. https://doi.org/10.1098/rspb.1958.0071
  26. Li H, Gu Y, Zhang Y, Lucas MJ, Wang Y. High glucose levels downregulate glucose transporter expression that correlates with increased oxidative stress in placental trophoblast cells in vitro. J Soc Gynecol Invest 2004;11:75-81. https://doi.org/10.1016/j.jsgi.2003.08.002
  27. Gross A. BCL-2 proteins: regulators of the mitochondrial apoptotic program. IUBMB Life 2001;52:231-6. https://doi.org/10.1080/15216540152846046
  28. Oswiecimska J, Suwala A, Swietochowska E, et al. Serum omentin levels in adolescent girls with anorexia nervosa and obesity. Physiol Res 2015;64:701-9.
  29. Vinals F, Gross A, Testar X, et al. High glucose concentrations inhibit glucose phosphorylation, but not glucose transport, in human endothelial cells. Biochim Biophys Acta (BBA)-Mol Cell Res 1999;1450: 119-29. https://doi.org/10.1016/S0167-4889(99)00035-X
  30. Sun Z, Shushanov S, LeRoith D, Wood TL. Decreased IGF type 1 receptor signaling in mammary epithelium during pregnancy leads to reduced proliferation, alveolar differentiation, and expression of insulin receptor substrate (IRS)-1 and IRS-2. Endocrinology 2011;152:3233-45. https://doi.org/10.1210/en.2010-1296
  31. Kleinberg DL, Barcellos-Hoff MH. The pivotal role of insulin-like growth factor I in normal mammary development. Endocrinol Metab Clin North Am 2011;40:461-71, vii. https://doi.org/10.1016/j.ecl.2011.06.001
  32. Shushanov SS. Insulin-like growth factors 1 and 2 regulate expression of ${\beta}$-casein in vitro in mouse mammary epithelial cells. Bull Exp Biol Med 2011;152:202-5. https://doi.org/10.1007/s10517-011-1488-4
  33. Lollivier V, Marnet PG, Delpal S, et al. Oxytocin stimulates secretory processes in lactating rabbit mammary epithelial cells. J Physiol 2006;570:125-40. https://doi.org/10.1113/jphysiol.2005.097816
  34. Ginger MR, Grigor MR. Comparative aspects of milk caseins. Comp Biochem Physiol B Biochem Mol Biol 1999;124:133-45. https://doi.org/10.1016/S0305-0491(99)00110-8

Cited by

  1. Moringa Extract Attenuates Inflammatory Responses and Increases Gene Expression of Casein in Bovine Mammary Epithelial Cells vol.9, pp.7, 2017, https://doi.org/10.3390/ani9070391