Lactogenic hormones regulate mammary protein synthesis in bovine mammary epithelial cells via the mTOR and JAK–STAT signal pathways
Q. Tian A B , H. R. Wang A E , M. Z. Wang A , C. Wang C and S. M. Liu DA College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
B Department of Food and Nutrition, Jiangsu Food Science College, Huaian 223003, China.
C School of Clinical Medicine, Jiangsu University, Zhenjiang 212013, China.
D School of Animal Biology of Faculty of Science, The University of Western Australia, Crawley, WA 6009, Australia.
E Corresponding author. Email: hrwang@yzu.edu.cn
Animal Production Science 56(11) 1803-1809 https://doi.org/10.1071/AN14113
Submitted: 25 February 2014 Accepted: 15 May 2015 Published: 21 July 2015
Abstract
The expression of CSN3, hormone receptor, the expression of genes regulating the mTOR, JAK–STAT signal pathways, and the relative content of к-casein as well as total casein were determined in the present study to explore the mechanism of the effect of lactogenic hormones on milk-protein synthesis in bovine mammary epithelial cells. The results showed that apoptosis of the cells was increased by inhibitor LY294002, while the expressions of genes encoding PKB, Rheb, PRAS40 and S6K1 in the mTOR signal pathway, JAK2, STAT5A in the JAK–STAT signal pathway, and genes encoding INSR, PRLR, NR3C1 and CSN3 were all downregulated, and the relative contents of κ-casein and total casein were decreased in the mammary epithelial cells compared with those in the control group. Comparatively, the inhibitory effects of AG-490 were more profound than those of LY294002, and the double block using both inhibitors had a greater effect than the single block. The CSN3 gene expression was downregulated and the content of milk casein was decreased by the inhibitors. In addition, the expression of the hormone receptor genes was downregulated. Our results suggest that lactogenic hormones, via their receptors in the membrane, regulated the JAK–STAT and m-TOR signal pathways, and affected cell proliferation and apoptosis, leading to changes in milk-protein synthesis.
Additional keywords: apoptosis, casein, gene expression, inhibitor, proliferation.
References
Akers RM (2006) Major advances associated with hormone and growth factor regulation of mammary growth and lactation in dairy cows. Journal of Dairy Science 89, 1222–1234.| Major advances associated with hormone and growth factor regulation of mammary growth and lactation in dairy cows.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xjt1SjsLs%3D&md5=7ae302e14b89e37cad5f3a939c65c7e1CAS | 16537955PubMed |
Ali S, Edery M, Pellegrini I, Lesueur L, Paly J, Djiane J, Kelly PA (1992) The Nb2 form of prolactin receptor is able to activate a milk protein gene promoter. Molecular Endocrinology (Baltimore, Md.) 6, 1242–1248.
Anderson SM, Rudolph MC, McManaman JL, Neville MC (2007) Secretory activation in the mammary gland: it’s not just about milk protein synthesis! Breast Cancer Research 9, 204
| Secretory activation in the mammary gland: it’s not just about milk protein synthesis!Crossref | GoogleScholarGoogle Scholar | 17338830PubMed |
Bossaert P, Cock HD, Leroy JLMR (2010) Immunohistochemical visualization of insulin receptors in formalin-fixed bovine ovaries post mortem and in granulosa cells collected in vivo. Theriogenology 73, 1210–1219.
| Immunohistochemical visualization of insulin receptors in formalin-fixed bovine ovaries post mortem and in granulosa cells collected in vivo.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXlvVWgu7Y%3D&md5=222599980f69ce3218d6f191f35106e4CAS | 20226514PubMed |
Boutin JM, Jolicoeur C, Okamura H, Gagnon J, Marc E, Shirota M, Banville D, Fourt ID, Djiane J, Kelly PA (1988) Cloning and expression of the rat prolactin receptor, a member of the growth hormone/prolactin receptor gene family. Cell 53, 69–77.
| Cloning and expression of the rat prolactin receptor, a member of the growth hormone/prolactin receptor gene family.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1cXlsVelsr0%3D&md5=2f7fe4fe42c32eeaec528010f4d2ced6CAS | 2832068PubMed |
Burgos SA, Dai M, Cant JP (2010) Nutrient availability and lactogenic hormones regulate mammary protein synthesis through the mammalian target of rapamycin signaling pathway. Journal of Dairy Science 93, 153–161.
| Nutrient availability and lactogenic hormones regulate mammary protein synthesis through the mammalian target of rapamycin signaling pathway.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhs1WhtbzJ&md5=f7e88ca4527e5b704f6d15673a3a29e5CAS | 20059914PubMed |
Chanat E, Martin P, Ollivier-Bousquet M (1999) Alpha(S1)-casein is required for the efficient transport of beta- and kappa-casein from the endoplasmic reticulum to the Golgi apparatus of mammary epithelial cells. Journal of Cell Science 112, 3399–3412.
Chen CC, Boxer RB, Stairs DB (2010) Akt is required for Stat5 activation and mammary differentiation. Breast Cancer 12, R72
Choi KM, Barash I, Rhoads AE (2004) Insulin and prolactin synergistically stimulate β-casein messenger ribonucleic acid translation by cytoplasmic polyadenylation. Molecular Endocrinology 18, 1670–1686.
| Insulin and prolactin synergistically stimulate β-casein messenger ribonucleic acid translation by cytoplasmic polyadenylation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXls1aktb0%3D&md5=acc1b1e976dbdc75f74ee34df174da65CAS | 15071091PubMed |
David M, Petricoin EF, Igarashi K, Feldman GM, Finbloom DS, Larner AC (1994) Prolactin activates the interferon-regulated P91 transcription factor and the JAK2 kinase by tyrosine phosphorylation. Proceedings of the National Academy of Sciences, USA 91, 7174–7178.
| Prolactin activates the interferon-regulated P91 transcription factor and the JAK2 kinase by tyrosine phosphorylation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXltVSht78%3D&md5=7ccadfcab5e7fbbe9c86f46c0808435cCAS |
Desrivières S, Kunz C, Barash I (2006) The biological functions of the versatile transcription factors STAT3 and STAT5 and new strategies for their targeted inhibition. Journal of Mammary Gland Biology and Neoplasia 11, 75–87.
| The biological functions of the versatile transcription factors STAT3 and STAT5 and new strategies for their targeted inhibition.Crossref | GoogleScholarGoogle Scholar | 16947086PubMed |
Dufner A, Thomas G (1999) Ribosomal S6 kinase signaling and the control of translation. Experimental Cell Research 253, 100–109.
| Ribosomal S6 kinase signaling and the control of translation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXnsF2itrc%3D&md5=856a07542d7f765f18aa60f51ec792a0CAS | 10579915PubMed |
Dusanter-Fourt I, Muller O, Ziemiecki A, Mayeux P, Drucker B, Djiane J, Wilks A, Harpur AG, Fisher S, Gisselbrecht S (1994) Identification of JAK protein kinases as signaling molecules for prolactin. Functional analysis of prolactin receptor and prolactin–erythropoietin receptor chimera expressed in lymphoid cells. The EMBO Journal 13, 2583–2591.
Edery M, Jolicoeur C, Levi-Meyrueis C, Dusanter-Fourt I, Pétridou B, Boutin JM, Lesueur L, Kelly PA, Djiane J (1989) Identification and sequence analysis of a second form of prolactin receptor by molecular cloning of complementary DNA from rabbit mammary gland. Proceedings of the National Academy of Sciences, USA 86, 2112–2116.
| Identification and sequence analysis of a second form of prolactin receptor by molecular cloning of complementary DNA from rabbit mammary gland.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3cXhslSkuw%3D%3D&md5=b5b9342b437b87c5ae92495a10f9cbebCAS |
Han Y, Watling D, Rogers NC, Stark GR (1997) JAK2 and STAT5, but not JAK1 and STAT1, are required for prolactin-induced beta-lactoglobulin transcription. Molecular Endocrinology 11, 1180–1188.
Hanigan MD, Cant JP, Weakley DC, Beckett JL (1998) An evaluation of postabsorptive protein and amino acid metabolism in the lactating dairy cow. Journal of Dairy Science 81, 3385–3401.
| An evaluation of postabsorptive protein and amino acid metabolism in the lactating dairy cow.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjvVantA%3D%3D&md5=0afe208461ce861e7ffd22ced019a5afCAS | 9891282PubMed |
Happ B, Groner B (1993) The activated mammary gland specific nuclear factor (MGF) enhances in vitro transcription of the beta-casein gene promoter. The Journal of Steroid Biochemistry and Molecular Biology 47, 21–30.
| The activated mammary gland specific nuclear factor (MGF) enhances in vitro transcription of the beta-casein gene promoter.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXpvVenug%3D%3D&md5=511dc0c0b11755a4700f0935de09909eCAS | 8274437PubMed |
Harir N, Boudot C, Friedbichler K (2008) Oncogenic kit controls neoplastic mast cell growth through a Stat5/PI3-kinase signaling cascade. Blood 112, 2463–2473.
| Oncogenic kit controls neoplastic mast cell growth through a Stat5/PI3-kinase signaling cascade.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhtFCru7%2FJ&md5=02c165824bf14ea420d3235c6b7d91d4CAS | 18579792PubMed |
Hay N, Sonenberg N (2004) Upstream and downstream of mTOR. Genes & Development 18, 1926–1945.
| Upstream and downstream of mTOR.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXmvFKqsLk%3D&md5=ba9055bdc69ddfd0d330cf378abb04b2CAS |
Lacasse P, Lollivier V, Dessauge F (2012) New developments on the galactopoietic role of prolactin in dairy ruminants. Domestic Animal Endocrinology 43, 154–160.
| New developments on the galactopoietic role of prolactin in dairy ruminants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XpvFOrtr0%3D&md5=194c691ad8b33682631886951652e27fCAS | 22281117PubMed |
Lebrun JJ, Ali S, Sofer L, Ullrich A, Kelly PA (1994) Prolactin-induced proliferation of Nb2 cells involves tyrosine phosphorylation of the prolactin receptor and its associated tyrosine kinase JAK2. The Journal of Biological Chemistry 269, 14021–14026.
Lebrun JJ, Ali S, Goffin V, Ullrich A, Kelly PA (1995) A single phosphotyrosine residue of the prolactin receptor is responsible for activation of gene transcription. Proceedings of the National Academy of Sciences, USA 92, 4031–4035.
| A single phosphotyrosine residue of the prolactin receptor is responsible for activation of gene transcription.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXlsVSksbo%3D&md5=302f9f455569b333c7389ebf0876da05CAS |
Lesueur L, Edery M, Ali S, Paly J, Kelly PA, Djiane J (1991) Comparison of long and short forms of the prolactin receptor on prolactin-induced milk protein gene transcription. Proceedings of the National Academy of Sciences, USA 88, 824–828.
| Comparison of long and short forms of the prolactin receptor on prolactin-induced milk protein gene transcription.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3MXpsFCntA%3D%3D&md5=9ca4632dbff142a4567ab7c3a7448c7eCAS |
Li S, Rosen JM (1995) Nuclear factor I and mammary gland factor (STAT5) play a critical role in regulating rat whey acidic protein gene expression in transgenic mice. Molecular and Cellular Biology 15, 2063–2070.
Manjarín R, Steibel JP, Kirkwood RN, Taylor NP, Trottier NL (2011) Transcript abundance of hormone receptors, mammalian target of rapamycin pathway-related kinases, insulin-like growth factor I, and milk proteins in porcine mammary tissue. Journal of Animal Science 1, 221–230.
Menzies KK, Lefevre C, Macmillan KL (2009) Insulin regulates milk protein synthesis at multiple levels in the bovine mammary gland. Functional & Integrative Genomics 9, 197–217.
| Insulin regulates milk protein synthesis at multiple levels in the bovine mammary gland.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXjsFyrsrk%3D&md5=8d51e144f3a4b77af1bfbe6a97110519CAS |
Pauloin A, Chanat E (2012) Prolactin and epidermal growth factor stimulate adipophilin synthesis in HC11 mouse mammary epithelial cells via the PI3-kinase/Akt/mTOR pathway. Biochimica et Biophysica Acta 1823, 987–996.
Qian L, Lopez V, Seo YA, Kelleher SL (2009) Prolactin regulates ZNT2 expression through the JAK2/STAT5 signaling pathway in mammary cells. American Journal of Physiology. Cell Physiology 297, C369–C377.
| Prolactin regulates ZNT2 expression through the JAK2/STAT5 signaling pathway in mammary cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtVahsbjM&md5=a1335b5b6a75e23921c1837e4cf47959CAS | 19494234PubMed |
Roehrborn CG, Siegel RL (1996) Safety and efficacy of doxazosin in benign prostatic hyperplasia: a pooled analysis of three double-blind, placebo-controlled studies. Urology 48, 406–415.
| Safety and efficacy of doxazosin in benign prostatic hyperplasia: a pooled analysis of three double-blind, placebo-controlled studies.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DyaK28zpslKjsg%3D%3D&md5=b98625d216f8d50e30dfe3c6f0abd0f8CAS | 8804494PubMed |
Rui H, Kirken RA, Farrar WL (1994) Activation of receptor-associated tyrosine kinase JAK2 by prolactin. The Journal of Biological Chemistry 269, 5364–5368.
Sakamoto K, Creamer BA, Triplett AA (2007) The Janus kinase 2 is required for expression and nuclear accumulation of cyclin D1 in proliferating mammary epithelial cells. Molecular Endocrinology 21, 1877–1892.
| The Janus kinase 2 is required for expression and nuclear accumulation of cyclin D1 in proliferating mammary epithelial cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXos12rsrg%3D&md5=86012334b5e30ec3e3fceb85d5d5bef3CAS | 17519353PubMed |
Santos MR, Yoon S, Matsuzaki Y, Mulligan RC, Melton DA (2001) Stemness. Transcriptional Profiling of Embryonic and Adult Stem Cells. Science 5593, 597–600.
Schwertfeger KL, Richert MM, Anderson SM (2001) Mammary gland involution is delayed by activated Akt in transgenic mice. Molecular Endocrinology 15, 867–881.
| Mammary gland involution is delayed by activated Akt in transgenic mice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXjvFKjs7g%3D&md5=ed47d44012b40eb1329781902d2c4922CAS | 11376107PubMed |
Schwertfeger KL, McManaman JL, Palmer CA, Neville MC, Ander-son SM (2003) Expression of constitutively activated Akt in the mammary gland leads to excess lipid synthesis during pregnancy and lactation. Journal of Lipid Research 44, 1100–1112.
| Expression of constitutively activated Akt in the mammary gland leads to excess lipid synthesis during pregnancy and lactation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXltFarsrg%3D&md5=6099f3bc3815c45a541c7baf6446cb1eCAS | 12700340PubMed |
Shirota M, Banville D, Ali S, Jolicoeur C, Boutin JM, Edery M, Djiane J, Kelly PA (1990) Expression of two forms of prolactin receptor in rat ovary and liver. Molecular Endocrinology 4, 1136–1143.
| Expression of two forms of prolactin receptor in rat ovary and liver.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3MXmsV2hs78%3D&md5=7dd479a428e90197e10aa5f661dc1637CAS | 2293022PubMed |
Tian Q, Wang HR (2013) Effects of insulin, prolactin and hydrocortisone on cell proliferation and apoptosis in bovine mammary epithelial cells. Chinese Journal of Feed 2, 8–12.
Tian Q, Ji Y, Pang XY (2013a) Effects of insulin on growth and expressions of κ-casein and insulin receptor genes in bovine mammary epithelial cells. Chinese Journal of Animal Nutrition 25, 77–87.
Tian Q, Ji Y, Pang XY (2013b) The action mechanism of insulin effects on the expression of CSN1S1 mRNA in mammary gland epithelial cells of Holstein cows in vitro. Chinese Journal of Animal Nutrition 25, 550–560.
Tokudome T, Horio T, Yoshihara F, Suga S, Kawano Y, Kohno M (2004) Direct effects of high glucose and insulin on protein synthesis in cultured cardiac myocytes and DNA and collagen synthesis in cardiac fibroblasts. Metabolism: Clinical and Experimental 53, 710–715.
| Direct effects of high glucose and insulin on protein synthesis in cultured cardiac myocytes and DNA and collagen synthesis in cardiac fibroblasts.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXkt1Cmsr4%3D&md5=7ad354a41ac11c2ff6fbaca752f567d5CAS |
Wang B, Xiao Z, Chen B (2008) Nogo-66 promotes the differentiation of neural progenitors into astroglial lineage cells through mTOR–STAT3 pathway. Public Library of Science One 3, e1856
Waters MJ, Daniel N, Bignon C, Djiane J (1995) The rabbit mammary gland prolactin receptor is tyrosine-phosphorylated in response to prolactin in vivo and in vitro. The Journal of Biological Chemistry 270, 5136–5143.
| The rabbit mammary gland prolactin receptor is tyrosine-phosphorylated in response to prolactin in vivo and in vitro.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXksVSlt78%3D&md5=2ba4011b428dfce70c5ce2471de44861CAS | 7534288PubMed |
Xie J, LeBaron MJ, Nevalainen MT, Rui H (2002) Role of tyrosine kinase Jak2 in prolactin-induced differentiation and growth of mammary epithelial cells. The Journal of Biological Chemistry 277, 14020–14030.
| Role of tyrosine kinase Jak2 in prolactin-induced differentiation and growth of mammary epithelial cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XjsFWnt74%3D&md5=116b43ade1962cf6b13a2b64c6f88920CAS | 11821424PubMed |
Yu Y, Ren W, Ren B (2009) Expression of signal transducers and activator of transcription 3 (STAT3) determines differentiation of olfactory bulb cells. Molecular and Cellular Biology 320, 101–108.
Zhou Y, Akers RM, Jiang H (2008) Growth hormone can induce expression of four major milk protein genes in transfected MAC-T cells. Journal of Dairy Science 91, 100–108.
| Growth hormone can induce expression of four major milk protein genes in transfected MAC-T cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsVCrtQ%3D%3D&md5=6bc5b95f61143fd5b8dbc70020bbef97CAS | 18096930PubMed |