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BMC Cancer
Published in: BMC Cancer 1/2015

Open Access 01-12-2015 | Research article

Neoplastic transformation of porcine mammary epithelial cells in vitro and tumor formation in vivo

Authors: A. R. Rowson-Hodel, R. Manjarin, J. F. Trott, R. D. Cardiff, A. D. Borowsky, R. C. Hovey

Published in: BMC Cancer | Issue 1/2015

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Abstract

Background

The mammary glands of pigs share many functional and morphological similarities with the breasts of humans, raising the potential of their utility for research into the mechanisms underlying normal mammary function and breast carcinogenesis. Here we sought to establish a model for the efficient manipulation and transformation of porcine mammary epithelial cells (pMEC) in vitro and tumor growth in vivo.

Methods

We utilized a vector encoding the red florescent protein tdTomato to transduce populations of pMEC from Yorkshire –Hampshire crossbred female pigs in vitro and in vivo. Populations of primary pMEC were then separated by FACS using markers to distinguish epithelial cells (CD140a-) from stromal cells (CD140a+), with or without further enrichment for basal and luminal progenitor cells (CD49f+). These separated pMEC populations were transduced by lentivirus encoding murine polyomavirus T antigens (Tag) and tdTomato and engrafted to orthotopic or ectopic sites in immunodeficient NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NSG) mice.

Results

We demonstrated that lentivirus effectively transduces pMEC in vitro and in vivo. We further established that lentivirus can be used for oncogenic-transformation of pMEC ex vivo for generating mammary tumors in vivo. Oncogenic transformation was confirmed in vitro by anchorage-independent growth, increased cell proliferation, and expression of CDKN2A, cyclin A2 and p53 alongside decreased phosphorylation of Rb. Moreover, Tag-transformed CD140a- and CD140a-CD49f + pMECs developed site-specific tumors of differing histopathologies in vivo.

Conclusions

Herein we establish a model for the transduction and oncogenic transformation of pMEC. This is the first report describing a porcine model of mammary epithelial cell tumorigenesis that can be applied to the study of human breast cancers.
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Literature
1.
Liu M and Hicklin D. Human tumor xenograft efficacy models. Tumor Models in Cancer Research. Cancer Drug Discovery and Development. Springer; 2011. p. 99-124.
2.
3.
4.
Hovey R, McFadden T, Akers R. Regulation of mammary gland growth and morphogenesis by the mammary fat pad: a species comparison. J Mammary Gland Biol Neoplasia. 1999;4:53–68.CrossRefPubMed
5.
6.
7.
8.
Turner C. The anatomy of the mammary gland of swine. In: Turner C, editor. The mammary gland. The anatomy of the udder of cattle and domestic animals. Columbia: Lucas Brothers; 1952. p. 279–314.
9.
10.
Kensinger RS, Collier RJ, Bazer FW, Ducsay CA, Becker HN. Nucleic acid, metabolic and histological changes in gilt mammary tissue during pregnancy and lactogenesis. J Anim Sci. 1982;54(6):1297–308.CrossRefPubMed
11.
12.
13.
14.
Ghajar CM, Bissell MJ. Extracellular matrix control of mammary gland morphogenesis and tumorigenesis: insights from imaging. Histochem Cell Biol. 2008;130(6):1105–18.CrossRefPubMedPubMedCentral
15.
Haslam SZ, Counterman L. Mammary stroma modulates hormonal responsiveness of mammary epithelium in vivo in the mouse. Endocrinology. 1991;129(4):2017–23.CrossRefPubMed
16.
Naylor MJ, Ormandy CJ. Mouse strain‐specific patterns of mammary epithelial ductal side branching are elicited by stromal factors. Dev Dynam. 2002;225(1):100–5.CrossRef
17.
Russo J, Gusterson B, Rogers A, Russo I, Wellings S, van Zwieten M. Comparative study of human and rat mammary tumorigenesis. Lab Invest. 1990;62:244–78.PubMed
18.
19.
20.
Hovey RC, Auldist DE, Mackenzie DD, McFadden TB. Preparation of an epithelium-free mammary fat pad and subsequent mammogenesis in ewes. J Anim Sci. 2000;78(8):2177–85.CrossRefPubMed
21.
Stampfer MR, Yaswen P and Taylor-Papadimitriou J. Culture of Human Mammary Epithelial Cells. In: Freshney I and Freshney M, editors. Culture of Epithelial Cells. John Wiley & Sons: New York, USA; 2004
22.
Barde I, Salmon P, Trono D. Production and titration of lentiviral vectors. Curr Prot Neurosci. 2010;4:1–23.
23.
Byrne KM, Cheng X, Vierck J, Erickson S, Greene EA, Duckett S, et al. Use of a 96-well plate reader to evaluate proliferation of equine satellite cell clones in vitro. Methods Cell Sci. 1998;19(4):311–6.CrossRef
24.
Landua JD, Visbal AP, Lewis MT. Methods for preparing fluorescent and neutral red-stained whole mounts of mouse mammary glands. J Mammary Gland Biol Neoplasia. 2009;14(4):411–5.CrossRefPubMedPubMedCentral
25.
26.
Shi S-R, Cote RJ, Wu L, Liu C, Datar R, Shi Y, et al. DNA extraction from archival formalin-fixed, paraffin-embedded tissue sections based on the antigen retrieval principle: heating under the influence of pH. J Histochem Cytochem. 2002;50(8):1005–11. doi:10.​1177/​0022155402050008​02.CrossRefPubMed
27.
Trott JF, Freking BA, Hovey RC. Variation in the coding and 3′ untranslated regions of the porcine prolactin receptor short form modifies protein expression and function. Anim Gen. 2014;45(1):74–86.CrossRef
28.
Vanklompenberg MK, Manjarin R, Trott JF, McMicking HF, Hovey RC. Late gestational hyperprolactinemia accelerates mammary epithelial cell differentiation that leads to increased milk yield. J Anim Sci. 2013;91(3):1102–11. doi:10.​2527/​jas.​2012-5903.CrossRefPubMed
29.
Trott JF, Horigan KC, Gloviczki JM, Costa KM, Freking BA, Farmer C, et al. Tissue-specific regulation of porcine prolactin receptor expression by estrogen, progesterone, and prolactin. J Endocrinol. 2009;202(1):153–66. doi:10.​1677/​joe-08-0486.CrossRefPubMed
30.
31.
Sheffield LG, Welsch CW. Transplantation of human breast epithelia to mammary-gland‐free fat‐pads of athymic nude mice: Influence of mammotrophic hormones on growth of breast epithelia. Intl J Cancer. 1988;41(5):713–9.CrossRef
32.
33.
Cardiff RD. Validity of mouse mammary tumour models for human breast cancer: comparative pathology. Microsc Res Tech. 2001;52(2):224–30.CrossRefPubMed
34.
35.
36.
37.
38.
39.
Bao L, Matsumura Y, Baban D, Sun Y, Tarin D. Effects of inoculation site and Matrigel on growth and metastasis of human breast cancer cells. Brit J Cancer. 1994;70(2):228.CrossRefPubMedPubMedCentral
40.
41.
Casey HW, Giles RC, Kwapien RP. Mammary neoplasia in animals: pathologic aspects and the effects of contraceptive steroids. In: Lingeman C, editor. Carcinogenic hormones. Recent results in cancer research/Fortschritte der Krebsforschung/Progrès dans les recherches sur le cancer. Berlin Heidelberg: Springer; 1979. p. 129–60.
42.
43.
Ronnov-Jessen L, Petersen O, Bissell M. Cellular changes involved in conversion of normal to malignant breast: The importance of the stromal reaction. Physiol Rev. 1996;76:69–125.PubMed
44.
Adam S, Rund L, Kuzmuk K, Zachary J, Schook L, Counter C. Genetic induction of tumorigenesis in swine. Oncogene. 2006;26(7):1038–45.CrossRefPubMed
45.
46.
Schüchner S, Wintersberger E. Binding of polyomavirus small T antigen to protein phosphatase 2A is required for elimination of p27 and support of S-phase induction in concert with large T antigen. J Virol. 1999;73(11):9266–73.PubMedPubMedCentral
47.
Ozbun MA, Butel JS. Tumor suppressor p53 mutations and breast cancer: a critical analysis. Adv Cancer Res. 1995;66:71–141.CrossRefPubMed
48.
Varley J, Armour J, Swallow JE, Jeffreys AJ, Ponder BA, T’Ang A, et al. The retinoblastoma gene is frequently altered leading to loss of expression in primary breast tumours. Oncogene. 1989;4(6):725–9.PubMed
49.
50.
Dabrosin C, Palmer K, Muller WJ, Gauldie J. Estradiol promotes growth and angiogenesis in polyoma middle T transgenic mouse mammary tumor explants. Breast Cancer Res Treat. 2003;78(1):1–6.CrossRefPubMed
51.
52.
Land H, Parada LF, Weinberg RA. Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature. 1982;304(5927):596–602.CrossRef
53.
Guy CT, Cardiff RD, Muller WJ. Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol. 1992;12(3):954–61.CrossRefPubMedPubMedCentral
54.
55.
Rosner A, Miyoshi K, Landesman-Bollag E, Xu X, Seldin DC, Moser AR, et al. Pathway pathology: histological differences between ErbB/Ras and Wnt pathway transgenic mammary tumors. Am J Pathol. 2002;161(3):1087–97.CrossRefPubMedPubMedCentral
56.
Metadata
Title
Neoplastic transformation of porcine mammary epithelial cells in vitro and tumor formation in vivo
Authors
A. R. Rowson-Hodel
R. Manjarin
J. F. Trott
R. D. Cardiff
A. D. Borowsky
R. C. Hovey
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2015
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-015-1572-7

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