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Effect of growth factors and lactogenic hormones on expression of plasminogen activator-related genes and cell proliferation in a bovine mammary epithelial cell line

Published online by Cambridge University Press:  21 July 2011

Georgios Theodorou ,
Chiara Pecorini ,
Raffaella Rebucci ,
Francesca Saccone ,
Christina Lecchi ,
Ioannis Politis  and
Antonella Baldi
Georgios Theodorou*
Affiliation:
Department of Animal Science and Aquaculture, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece Department of Veterinary Science and Technologies for Food Safety, Università degli Studi di Milano, via Celoria 10, Milan, Italy
Chiara Pecorini
Affiliation:
Department of Veterinary Science and Technologies for Food Safety, Università degli Studi di Milano, via Celoria 10, Milan, Italy Centro interdipartimentale di studi Ghiandola mammaria (CISMA), Università degli Studi di Milano, Via Celoria 10, Milano, Italy
Raffaella Rebucci
Affiliation:
Department of Veterinary Science and Technologies for Food Safety, Università degli Studi di Milano, via Celoria 10, Milan, Italy
Francesca Saccone
Affiliation:
Department of Veterinary Science and Technologies for Food Safety, Università degli Studi di Milano, via Celoria 10, Milan, Italy
Christina Lecchi
Affiliation:
Department of Animal Pathology, Hygiene and Veterinary Public Health, Università degli Studi di Milano, via Celoria 10, Milan, Italy
Ioannis Politis*
Affiliation:
Department of Animal Science and Aquaculture, Agricultural University of Athens, 75 Iera Odos, 11855 Athens, Greece
Antonella Baldi
Affiliation:
Department of Veterinary Science and Technologies for Food Safety, Università degli Studi di Milano, via Celoria 10, Milan, Italy Centro interdipartimentale di studi Ghiandola mammaria (CISMA), Università degli Studi di Milano, Via Celoria 10, Milano, Italy
*
*For correspondence; e-mail: [email protected]
*For correspondence; e-mail: [email protected]
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Abstract

There is conflicting evidence in the literature as to whether up-regulation of urokinase plasminogen activator (u-PA) expression is related to bovine mammary epithelial cell growth. The role of u-PA receptor (u-PAR) and that of the plasminogen activator inhibitors type 1 and type 2 (PAI-1 and PAI-2) in bovine mammary epithelial cell proliferation is not known. The effect of growth factors and various hormones known to affect mammary function on expression of u-PA, u-PAR, PAI-1, PAI-2 and cell proliferation using the BME-UV1 bovine mammary epithelial cell line was examined. Cell proliferation was measured using the MTT assay and direct cell enumeration. Results showed that both IGF-1 and EGF increased cell proliferation but EGF was a more potent mitogen than IGF-1. Furthermore, IGF-1 increased by 2-fold expression of both u-PA and u-PAR while EGF increased by 3·8-fold the expression of only u-PAR. Both growth factors had no effect on expression of PAI-1 and PAI-2. In a manner consistent with changes in gene expression, EGF and to a lesser extent IGF-1 up-regulated total cell associated, membrane-bound and secreted u-PA activity. Thus, a strong correlation exists between u-PAR gene expression along with the activity of u-PA present on cell membranes and cell proliferation. Dexamethasone, prolactin and surprisingly insulin had no effect on cell proliferation. Dexamethasone alone and when combined with insulin or prolactin up-regulated gene expression of both PAI- and PAI-2 but not that of u-PA and u-PAR. Decreased total cell-associated, membrane-bound and secreted u-PA activity was detected in cells cultured in the presence of dexamethasone when combined with insulin or prolactin. However no such effect was observed in the presence of dexamethasone alone. Thus, dexamethasone acting synergistically with prolactin or insulin inhibits the activation of the plasmin-plasminogen system but this inhibition is not correlated with any changes in cell proliferation.

Keywords

Growth factorslactogenic hormonesplasminogen activator-related genesmammary proliferation
Type
Research Article
Information
Journal of Dairy Research , Volume 78 , Issue 3 , August 2011 , pp. 365 - 372
Copyright
Copyright © Proprietors of Journal of Dairy Research 2011

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References

Accornero, P, Martignani, E, Miretti, S, Cucuzza, LS & Baratta, M 2009 Epidermal growth factor and hepatocyte growth factor receptors collaborate to induce multiple biological responses in bovine mammary epithelial cells. Journal of Dairy Science 92 3667–3675CrossRefGoogle ScholarPubMed
Ahmad, A, Kong, D, Wang, Z, Sarkar, SH, Banerjee, S & Sarkar, FH 2009 Down-regulation of uPA and uPAR by 3,3′-diindolylmethane contributes to the inhibition of cell growth and migration of breast cancer cells. Journal of Cell Biochemistry 108 916–925CrossRefGoogle Scholar
Akers, RM 1985 Lactogenic hormones: binding sites, mammary growth, secretory cell differentiation, and milk biosynthesis in ruminants. Journal of Dairy Science 68 501–519CrossRefGoogle ScholarPubMed
Alfano, D, Franco, P, Vocca, I, Gambi, N, Pisa, V, Mancini, A, Caputi, M, Carriero, MV, Iaccarino, I & Stoppelli, MP 2005 The urokinase plasminogen activator and its receptor: role in cell growth and apoptosis. Thrombosis and Haemostasis 93 205–211CrossRefGoogle ScholarPubMed
Baldi, A, Losio, MN, Cheli, F, Rebucci, R, Sangalli, L, Fusi, E, Bertasi, B, Pavoni, E, Carli, S & Politis, I 2004 Evaluation of the protective effects of alpha-tocopherol and retinol against ochratoxin A cytotoxicity. British Journal of Nutrition 91 507–512CrossRefGoogle ScholarPubMed
Baumrucker, CR & Stemberger, BH 1989 Insulin and insulin-like growth factor-I stimulate DNA synthesis in bovine mammary tissue in vitro. Journal of Animal Science 67 3503–3514CrossRefGoogle ScholarPubMed
Busso, N, Belin, D, Failly-Crepin, C & Vassalli, JD 1987 Glucocorticoid modulation of plasminogen activators and of one of their inhibitors in the human mammary carcinoma cell line MDA-MB-231. Cancer Research 47 364–370Google ScholarPubMed
Cheli, F, Politis, I, Rossi, L, Fusi, E & Baldi, A 2003 Effects of retinoids on proliferation and plasminogen activator expression in a bovine mammary epithelial cell line. Journal of Dairy Research 70 367–372CrossRefGoogle Scholar
Doll, F, Pfeilschifter, J & Huwiler, A 2007 Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration. Endocrine Related Cancer 14 325–335CrossRefGoogle ScholarPubMed
Dunn, SE, Torres, JV, Nihei, N & Barrett, JC 2000 The insulin-like growth factor-1 elevates urokinase-type plasminogen activator-1 in human breast cancer cells: a new avenue for breast cancer therapy. Molecular Carcinogenesis 27 10–173.0.CO;2-Z>CrossRefGoogle Scholar
Dunn, SE, Torres, JV, Oh, JS, Cykert, DM & Barrett, JC 2001 Up-regulation of urokinase-type plasminogen activator by insulin-like growth factor-I depends upon phosphatidylinositol-3 kinase and mitogen-activated protein kinase kinase. Cancer Research 61 1367–1374Google ScholarPubMed
Gandhari, M, Arens, N, Majety, M, Dorn-Beineke, A & Hildenbrand, R 2006 Urokinase-type plasminogen activator induces proliferation in breast cancer cells. International Journal of Oncology 28 1463–1470Google ScholarPubMed
Henderson, BR & Kefford, RF 1993 Dexamethasone decreases urokinase plasminogen activator mRNA stability in MAT 13762 rat mammary carcinoma cells. British Journal of Cancer 67 99–101CrossRefGoogle ScholarPubMed
Jo, M, Thomas, KS, Takimoto, S, Gaultier, A, Hsieh, EH, Lester, RD & Gonias, SL 2007 Urokinase receptor primes cells to proliferate in response to epidermal growth factor. Oncogene 26 2585–2594CrossRefGoogle ScholarPubMed
Lipka, C, Mankertz, J, Fromm, M, Lubbert, H, Buhler, H, Kuhn, W, Ragosch, V & Hundertmark, S 2004 Impairment of the antiproliferative effect of glucocorticosteroids by 11beta-hydroxysteroid dehydrogenase type 2 overexpression in MCF-7 breast-cancer cells. Hormone Metabolism Research 36 437–444CrossRefGoogle ScholarPubMed
Marshall, OJ 2004 PerlPrimer: cross-platform, graphical primer design for standard, bisulphite and real-time PCR. Bioinformatics 20 2471–2472CrossRefGoogle ScholarPubMed
Olazabal, I, Munoz, J, Ogueta, S, Obregon, E & Garcia-Ruiz, JP 2000 Prolactin (PRL)-PRL receptor system increases cell proliferation involving JNK (c-Jun amino terminal kinase) and AP-1 activation: inhibition by glucocorticoids. Molecular Endocrinology 14 564–575CrossRefGoogle ScholarPubMed
Politis, I 1996 Plasminogen activator system: implications for mammary cell growth and involution. Journal of Dairy Science 79 1097–1107CrossRefGoogle ScholarPubMed
Politis, I, Zavizion, B, White, JH, Goldberg, JJ, Baldi, A & Akers, RM 1995 Hormonal and extracellular matrix regulation of plasminogen activator in a bovine mammary epithelial cell line. Endocrine 3 345–350CrossRefGoogle Scholar
Purup, S, Vestergaard, M & Sejrsen, K 2000 Involvement of growth factors in the regulation of pubertal mammary growth in cattle. Advances in Experimental Medicine and Biology 480 27–43CrossRefGoogle ScholarPubMed
Rabot, A, Sinowatz, F, Berisha, B, Meyer, HH & Schams, D 2007 Expression and localization of extracellular matrix-degrading proteinases and their inhibitors in the bovine mammary gland during development, function, and involution. Journal of Dairy Science 90 740–748CrossRefGoogle ScholarPubMed
Rubis, B, Grodecka-Gazdecka, S, Lecybyl, R, Ociepa, M, Krozowski, Z & Trzeciak, WH 2004 Contribution of protein kinase A and protein kinase C signalling pathways to the regulation of HSD11B2 expression and proliferation of MCF-7 cells. Acta Biochimica Polonica 51 919–924Google Scholar
Sasaki, CY, Lin, H & Passaniti, A 1999 Regulation of urokinase plasminogen activator (uPA) activity by E-cadherin and hormones in mammary epithelial cells. Journal of Cell Physiology 181 1–133.0.CO;2-B>CrossRefGoogle ScholarPubMed
Stoppelli, MP, Tacchetti, C, Cubellis, MV, Corti, A, Hearing, VJ, Cassani, G, Appella, E & Blasi, F 1986 Autocrine saturation of pro-urokinase receptors on human A431 cells. Cell 45 675–684CrossRefGoogle ScholarPubMed
Zavizion, B, van Duffelen, M, Schaeffer, W & Politis, I 1996 Establishment and characterization of a bovine mammary epithelial cell line with unique properties. In Vitro Cellular & Developmental Biology – Animal 32 138–148CrossRefGoogle ScholarPubMed
Zhou, Y, Capuco, AV & Jiang, H 2008 Involvement of connective tissue growth factor (CTGF) in insulin-like growth factor-I (IGF1) stimulation of proliferation of a bovine mammary epithelial cell line. Domestic Animal Endocrinology 35 180–189CrossRefGoogle ScholarPubMed