CCL2-driven inflammation increases mammary gland stromal density and cancer susceptibility in a transgenic mouse model

Xuan Sun, Danielle J Glynn, Leigh J Hodson, Cecilia Huo, Kara Britt, Erik W Thompson, Lucy Woolford, Andreas Evdokiou, Jeffrey W Pollard, Sarah A Robertson, Wendy V Ingman, Xuan Sun, Danielle J Glynn, Leigh J Hodson, Cecilia Huo, Kara Britt, Erik W Thompson, Lucy Woolford, Andreas Evdokiou, Jeffrey W Pollard, Sarah A Robertson, Wendy V Ingman

Abstract

Background: Macrophages play diverse roles in mammary gland development and breast cancer. CC-chemokine ligand 2 (CCL2) is an inflammatory cytokine that recruits macrophages to sites of injury. Although CCL2 has been detected in human and mouse mammary epithelium, its role in regulating mammary gland development and cancer risk has not been explored.

Methods: Transgenic mice were generated wherein CCL2 is driven by the mammary epithelial cell-specific mouse mammary tumour virus 206 (MMTV) promoter. Estrous cycles were tracked in adult transgenic and non-transgenic FVB mice, and mammary glands collected at the four different stages of the cycle. Dissected mammary glands were assessed for cyclical morphological changes, proliferation and apoptosis of epithelium, macrophage abundance and collagen deposition, and mRNA encoding matrix remodelling enzymes. Another cohort of control and transgenic mice received carcinogen 7,12-Dimethylbenz(a)anthracene (DMBA) and tumour development was monitored weekly. CCL2 protein was also quantified in paired samples of human breast tissue with high and low mammographic density.

Results: Overexpression of CCL2 in the mammary epithelium resulted in an increased number of macrophages, increased density of stroma and collagen and elevated mRNA encoding matrix remodelling enzymes lysyl oxidase (LOX) and tissue inhibitor of matrix metalloproteinases (TIMP)3 compared to non-transgenic controls. Transgenic mice also exhibited increased susceptibility to development of DMBA-induced mammary tumours. In a paired sample cohort of human breast tissue, abundance of epithelial-cell-associated CCL2 was higher in breast tissue of high mammographic density compared to tissue of low mammographic density.

Conclusions: Constitutive expression of CCL2 by the mouse mammary epithelium induces a state of low level chronic inflammation that increases stromal density and elevates cancer risk. We propose that CCL2-driven inflammation contributes to the increased risk of breast cancer observed in women with high mammographic density.

Keywords: Chemokine (C-C motif) ligand 2; Development; Macrophage; Mammary gland; Mammographic density; Mouse model.

Figures

Fig. 1
Fig. 1
Generation of mouse mammary tumour virus 206 transgenic mice (Mmtv-Ccl2). The Mmtv-Ccl2 expression cassette was detected in three of twenty-nine mice by PCR screening and the expression of both endogenous genomic and cloned Ccl2 mRNA in different tissues was measured by RT-PCR with product sizes of 451 bp and 125 bp, respectively (a). CCL2 protein detected by immunohistochemistry in the mammary gland from female offspring from founder mouse #29 (b) compared to non-transgenic control (c). RNA encoding MMTV (d) and CCL2 (e), and CCL2 protein (f) were quantified in spleen (Sp), kidney (Kid), ovary (Ov), liver (Liv), salivary gland (SG) and mammary gland (MG) from Mmtv-Ccl2 and non-transgenic control mice. Abundance of mRNA was normalised to Actb expression, and is given in arbitrary units where the average of the non-transgenic mammary gland control is 1; n = 5 per group. Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test, *p < 0.05
Fig. 2
Fig. 2
The effect of CCL2 overexpression on macrophage abundance and location within and around mammary epithelium. Paraffin sections of mammary gland tissue from control and Mmtv-Ccl2 were stained with anti-macrophage-specific F4/80 antibody to detect macrophages in the stroma surrounding ductal (a, b) and alveolar epithelium (c, d) of mammary glands from adult control and Mmtv-Ccl2 mice. Macrophages are indicated by black arrows. The number of F4/80-positive macrophages was quantified and represented as F4/80-positive macrophages/mm2 (e); n = 6 per group. Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test, *p < 0.05
Fig. 3
Fig. 3
The effect of CCL2 overexpression on mammary gland morphogenesis during ovarian cycle. Mammary glands from control and Mmtv-Ccl2 mice were whole-mounted and stained with carmine alum at estrus (a, e), metestrus (b, f), diestrus (c, g) and proestrus (d, h), respectively. The number of branch points per millimetre was calculated (k). Sections of paraffin-embedded mammary glands from control and Mmtv-Ccl2 mice at the four stages of the cycle were H&E-stained (i, j, respectively, only proestrus stage shown) and alveolar epithelium quantified (l); n = 6–9 per group. Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test, *p < 0.05
Fig. 4
Fig. 4
The effect of CCL2 overexpression on mammary epithelial cell proliferation and cell death. Paraffin-embedded sections of mammary gland tissue from control and Mmtv-Ccl2 mice at proestrus were stained with anti-bromodeoxyuridine (BrdU) antibody to detect proliferating ductal (a and b) and alveolar (c and d) epithelial cells. The number of BrdU-positive cells (brown-stained cells) within ductal and alveolar epithelium was calculated and expressed as BrdU-positive cells/mm2 (i). Paraffin-embedded sections of mammary gland tissue from control and Mmtv-Ccl2 mice at proestrus were stained with TUNEL to detect dying ductal (e and f) and alveolar (g and h) epithelial cells. The percent of TUNEL-positive cells (green-stained cells) within ductal and alveolar epithelium was calculated (j); n = 6 per group. Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test, *p < 0.05
Fig. 5
Fig. 5
The effect of CCL2 overexpression on abundance of stroma and collagen. Sections of mammary gland tissue from control (a, d, f) and Mmtv-Ccl2 (b, e, h) mice at proestrus were stained with H&E (a, b), Masson’s trichrome (c, d) and collagen I antibody (g, h) and quantified (c, f, i); n = 6 per group. Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test, *p < 0.05 compared to control. Mammary glands from both groups of mice (n = 8) were dissected and frozen in liquid nitrogen. Messenger RNAs of collagen remodelling enzymes including Lox, Mmp2, Mmp9, Timp1, Timp2 and Timp3 were extracted and measured by RT-PCR. The amount of mRNA was normalised to Actb expression, and is given in arbitrary units, where the average of the control is 1 (j). Data are presented as mean + SEM with statistical analysis conducted using the unpaired t test; *p < 0.05 compared to control. LOX lysyl oxidase, MMP matrix metalloproteinase, TIMP tissue inhibitor of matrix metalloproteinases
Fig. 6
Fig. 6
The effect of CCL2 overexpression on mammary gland cancer susceptibility in mice. Kaplan–Meier survival plot showing the percentage of tumour-free mice following 7,12-Dimethylbenz(a)anthracene (DMBA) treatment in weeks for control and Mmtv-Ccl2 mice; n = 18 per group. In Mmtv-Ccl2 mice, the incidence of mammary tumours was increased (p = 0.04 chi squared) and mammary tumour-free survival was reduced (p = 0.025 log rank) compared to control mice
Fig. 7
Fig. 7
CCL2 in paired samples of high and low mammographic density (MD). CCL2 immunostaining of human breast tissue of low MD (a) and high MD (b) and isotype-matched negative control (c). Intensity of staining within the epithelium was quantified and data are presented as percent change in abundance of CCL2 in high density tissue compared to low density tissue for each paired sample; horizontal bar indicates the mean (d) analysis by paired sample t test (n = 13). HMD high mammographic density

References

    1. Chua AC, Hodson LJ, Moldenhauer LM, Robertson SA, Ingman WV. Dual roles for macrophages in ovarian cycle-associated development and remodelling of the mammary gland epithelium. Development. 2010;137(24):4229–38. doi: 10.1242/dev.059261.
    1. Ingman WV, Wyckoff J, Gouon-Evans V, Condeelis J, Pollard JW. Macrophages promote collagen fibrillogenesis around terminal end buds of the developing mammary gland. Dev Dyn. 2006;235(12):3222–9. doi: 10.1002/dvdy.20972.
    1. Pollard JW, Hennighausen L. Colony stimulating factor 1 is required for mammary gland development during pregnancy. Proc Natl Acad Sci U S A. 1994;91(20):9312–6. doi: 10.1073/pnas.91.20.9312.
    1. Schwertfeger KL, Rosen JM, Cohen DA. Mammary gland macrophages: pleiotropic functions in mammary development. J Mammary Gland Biol Neoplasia. 2006;11(3–4):229–38. doi: 10.1007/s10911-006-9028-y.
    1. O'Brien J, Martinson H, Durand-Rougely C, Schedin P. Macrophages are crucial for epithelial cell death and adipocyte repopulation during mammary gland involution. Development. 2012;139(2):269–75. doi: 10.1242/dev.071696.
    1. Gouon Evans V, Rothenberg ME, Pollard JW. Postnatal mammary gland development requires macrophages and eosinophils. Development. 2000;127(11):2269–82.
    1. Hodson LJ, Chua AC, Evdokiou A, Robertson SA, Ingman WV. Macrophage phenotype in the mammary gland fluctuates over the course of the estrous cycle and is regulated by ovarian steroid hormones. Biol Reprod. 2013;89(3):65. doi: 10.1095/biolreprod.113.109561.
    1. Sun X, Ingman WV. Cytokine networks that mediate epithelial cell-macrophage crosstalk in the mammary gland: implications for development and cancer. J Mammary Gland Biol Neoplasia. 2014;19(2):191–201. doi: 10.1007/s10911-014-9319-7.
    1. Yang J, Hawkins OE, Barham W, Gilchuk P, Boothby M, Ayers GD, Joyce S, Karin M, Yull FE, Richmond A. Myeloid IKKbeta promotes antitumor immunity by modulating CCL11 and the innate immune response. Cancer Res. 2014;74(24):7274–84. doi: 10.1158/0008-5472.CAN-14-1091.
    1. Diefenbach A, Jamieson AM, Liu SD, Shastri N, Raulet DH. Ligands for the murine NKG2D receptor: expression by tumor cells and activation of NK cells and macrophages. Nat Immunol. 2000;1(2):119–26. doi: 10.1038/77793.
    1. Dranoff G. Cytokines in cancer pathogenesis and cancer therapy. Nat Rev Cancer. 2004;4(1):11–22. doi: 10.1038/nrc1252.
    1. Mantovani A, Sozzani S, Locati M, Allavena P, Sica A. Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol. 2002;23(11):549–55. doi: 10.1016/S1471-4906(02)02302-5.
    1. Qian BZ, Pollard JW. Macrophage diversity enhances tumor progression and metastasis. Cell. 2010;141(1):39–51. doi: 10.1016/j.cell.2010.03.014.
    1. Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140(6):883–99. doi: 10.1016/j.cell.2010.01.025.
    1. Yadav A, Saini V, Arora S. MCP-1: chemoattractant with a role beyond immunity: a review. Clin Chim Acta. 2010;411(21–22):1570–9. doi: 10.1016/j.cca.2010.07.006.
    1. Deshmane SL, Kremlev S, Amini S, Sawaya BE. Monocyte chemoattractant protein-1 (MCP-1): an overview. J Interf Cytokine Res. 2009;29(6):313–26. doi: 10.1089/jir.2008.0027.
    1. Soria G, Ben-Baruch A. The inflammatory chemokines CCL2 and CCL5 in breast cancer. Cancer Lett. 2008;267(2):271–85. doi: 10.1016/j.canlet.2008.03.018.
    1. Ueno T, Toi M, Saji H, Muta M, Bando H, Kuroi K, Koike M, Inadera H, Matsushima K. Significance of macrophage chemoattractant protein-1 in macrophage recruitment, angiogenesis, and survival in human breast cancer. Clin Cancer Res. 2000;6(8):3282–9.
    1. Qian BZ, Li J, Zhang H, Kitamura T, Zhang J, Campion LR, Kaiser EA, Snyder LA, Pollard JW. CCL2 recruits inflammatory monocytes to facilitate breast-tumour metastasis. Nature. 2011;475(7355):222–5. doi: 10.1038/nature10138.
    1. Fujimoto H, Sangai T, Ishii G, Ikehara A, Nagashima T, Miyazaki M, Ochiai A. Stromal MCP-1 in mammary tumors induces tumor-associated macrophage infiltration and contributes to tumor progression. Int J Cancer. 2009;125(6):1276–84. doi: 10.1002/ijc.24378.
    1. Arendt LM, McCready J, Keller PJ, Baker DD, Naber SP, Seewaldt V, Kuperwasser C. Obesity promotes breast cancer by CCL2-mediated macrophage recruitment and angiogenesis. Cancer Res. 2013;73(19):6080–93. doi: 10.1158/0008-5472.CAN-13-0926.
    1. Lu X, Kang Y. Chemokine (C-C motif) ligand 2 engages CCR2+ stromal cells of monocytic origin to promote breast cancer metastasis to lung and bone. J Biol Chem. 2009;284(42):29087–96. doi: 10.1074/jbc.M109.035899.
    1. Lin SJ, Cawson J, Hill P, Haviv I, Jenkins M, Hopper JL, Southey MC, Campbell IG, Thompson EW. Image-guided sampling reveals increased stroma and lower glandular complexity in mammographically dense breast tissue. Breast Cancer Res Treat. 2011;128(2):505–16. doi: 10.1007/s10549-011-1346-0.
    1. Alowami S, Troup S, Al-Haddad S, Kirkpatrick I, Watson PH. Mammographic density is related to stroma and stromal proteoglycan expression. Breast Cancer Res. 2003;5(5):R129–135. doi: 10.1186/bcr622.
    1. Huo CW, Chew G, Hill P, Huang D, Ingman W, Hodson L, Brown KA, Magenau A, Allam AH, McGhee E, et al. High mammographic density is associated with an increase in stromal collagen and immune cells within the mammary epithelium. Breast Cancer Res. 2015;17:79. doi: 10.1186/s13058-015-0592-1.
    1. Boyd NF, Guo H, Martin LJ, Sun L, Stone J, Fishell E, Jong RA, Hislop G, Chiarelli A, Minkin S, et al. Mammographic density and the risk and detection of breast cancer. N Engl J Med. 2007;356(3):227–36. doi: 10.1056/NEJMoa062790.
    1. Ingman WV, Robker RL, Woittiez K, Robertson SA. Null mutation in transforming growth factor beta1 disrupts ovarian function and causes oocyte incompetence and early embryo arrest. Endocrinology. 2006;147(2):835–45. doi: 10.1210/en.2005-1189.
    1. Rutledge BJ, Rayburn H, Rosenberg R, North RJ, Gladue RP, Corless CL, Rollins BJ. High level monocyte chemoattractant protein-1 expression in transgenic mice increases their susceptibility to intracellular pathogens. J Immunol (Baltimore, Md: 1950) 1995;155(10):4838–43.
    1. Huang AL, Ostrowski MC, Berard D, Hager GL. Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus. Cell. 1981;27:245–55. doi: 10.1016/0092-8674(81)90408-6.
    1. Ingman WV, Robertson SA. Mammary gland development in transforming growth factor beta1 null mutant mice: systemic and epithelial effects. Biol Reprod. 2008;79(4):711–7. doi: 10.1095/biolreprod.107.067272.
    1. Fata JE, Chaudhary V, Khokha R. Cellular turnover in the mammary gland is correlated with systemic levels of progesterone and not 17beta-estradiol during the estrous cycle. Biol Reprod. 2001;65(3):680–8. doi: 10.1095/biolreprod65.3.680.
    1. Chua CLHL, Robertson SA, Ingman WV. Dual roles of macrophages in ovarian cycle-associated development and remodelling of the mammary gland epithelium. Development. 2010;137(24):4229–38. doi: 10.1242/dev.059261.
    1. Jasper MJ, Tremellen KP, Robertson SA. Primary unexplained infertility is associated with reduced expression of the T-regulatory cell transcription factor Foxp3 in endometrial tissue. Mol Hum Reprod. 2006;12(5):301–8. doi: 10.1093/molehr/gal032.
    1. Rollins BJ, Morrison ED, Stiles CD. Cloning and expression of JE, a gene inducible by platelet-derived growth factor and whose product has cytokine-like properties. Proc Natl Acad Sci U S A. 1988;85(11):3738–42. doi: 10.1073/pnas.85.11.3738.
    1. Kanda H, Tateya S, Tamori Y, Kotani K, Hiasa K, Kitazawa R, Kitazawa S, Miyachi H, Maeda S, Egashira K, et al. MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. J Clin Investig. 2006;116(6):1494–505. doi: 10.1172/JCI26498.
    1. Fuentes ME, Durham SK, Swerdel MR, Lewin AC, Barton DS, Megill JR, Bravo R, Lira SA. Controlled recruitment of monocytes and macrophages to specific organs through transgenic expression of monocyte chemoattractant protein-1. J Immunol (Baltimore, Md: 1950) 1995;155(12):5769–76.
    1. Lu B, Rutledge BJ, Gu L, Fiorillo J, Lukacs NW, Kunkel SL, North R, Gerard C, Rollins BJ. Abnormalities in monocyte recruitment and cytokine expression in monocyte chemoattractant protein 1-deficient mice. J Exp Med. 1998;187(4):601–8. doi: 10.1084/jem.187.4.601.
    1. Dasari P, Sharkey DJ, Noordin E, Glynn DJ, Hodson LJ, Chin PY, Evdokiou A, Robertson SA, Ingman WV. Hormonal regulation of the cytokine microenvironment in the mammary gland. J Reprod Immunol. 2014;106:58–66. doi: 10.1016/j.jri.2014.07.002.
    1. O'Brien J, Lyons T, Monks J, Lucia MS, Wilson RS, Hines L, Man Y, Borges V, Schedin P. Alternatively activated macrophages and collagen remodeling characterize the postpartum involuting mammary gland across species. Am J Pathol. 2010;176(3):1241–55. doi: 10.2353/ajpath.2010.090735.
    1. Glynn DJ, Hutchinson MR, Ingman WV. Toll-like receptor 4 regulates lipopolysaccharide-induced inflammation and lactation insufficiency in a mouse model of mastitis. Biol Reprod. 2014;90(5):91. doi: 10.1095/biolreprod.114.117663.
    1. Chua AC, Hodson LJ, Moldenhauer LM, Robertson SA, Ingman WV. Dual roles for macrophages in ovarian cycle-associated development and remodelling of the mammary gland epithelium. Development. 2010;137
    1. Maller O, Martinson H, Schedin P. Extracellular matrix composition reveals complex and dynamic stromal-epithelial interactions in the mammary gland. J Mammary Gland Biol Neoplasia. 2010;15(3):301–18. doi: 10.1007/s10911-010-9189-6.
    1. Maller O, Hansen KC, Lyons TR, Acerbi I, Weaver VM, Prekeris R, Tan AC, Schedin P. Collagen architecture in pregnancy-induced protection from breast cancer. J Cell Sci. 2013;126(Pt 18):4108–10. doi: 10.1242/jcs.121590.
    1. Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, Fong SF, Csiszar K, Giaccia A, Weninger W, et al. Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell. 2009;139(5):891–906. doi: 10.1016/j.cell.2009.10.027.
    1. Antoniades HN, Neville-Golden J, Galanopoulos T, Kradin RL, Valente AJ, Graves DT. Expression of monocyte chemoattractant protein 1 mRNA in human idiopathic pulmonary fibrosis. Proc Natl Acad Sci U S A. 1992;89(12):5371–5. doi: 10.1073/pnas.89.12.5371.
    1. Brieland JK, Jones ML, Flory CM, Miller GR, Warren JS, Phan SH, Fantone JC. Expression of monocyte chemoattractant protein-1 (MCP-1) by rat alveolar macrophages during chronic lung injury. Am J Respir Cell Mol Biol. 1993;9(3):300–5. doi: 10.1165/ajrcmb/9.3.300.
    1. Baran CP, Opalek JM, McMaken S, Newland CA, O'Brien JM, Jr, Hunter MG, Bringardner BD, Monick MM, Brigstock DR, Stromberg PC, et al. Important roles for macrophage colony-stimulating factor, CC chemokine ligand 2, and mononuclear phagocytes in the pathogenesis of pulmonary fibrosis. Am J Respir Crit Care Med. 2007;176(1):78–89. doi: 10.1164/rccm.200609-1279OC.
    1. Lloyd CM, Dorf ME, Proudfoot A, Salant DJ, Gutierrez-Ramos JC. Role of MCP-1 and RANTES in inflammation and progression to fibrosis during murine crescentic nephritis. J Leukoc Biol. 1997;62(5):676–80.
    1. Weitkamp B, Cullen P, Plenz G, Robenek H, Rauterberg J. Human macrophages synthesize type VIII collagen in vitro and in the atherosclerotic plaque. FASEB J. 1999;13(11):1445–57.
    1. Schnoor M, Cullen P, Lorkowski J, Stolle K, Robenek H, Troyer D, Rauterberg J, Lorkowski S. Production of type VI collagen by human macrophages: a new dimension in macrophage functional heterogeneity. J Immunol (Baltimore, Md: 1950) 2008;180(8):5707–19. doi: 10.4049/jimmunol.180.8.5707.
    1. Schmid-Kotsas A, Gross HJ, Menke A, Weidenbach H, Adler G, Siech M, Beger H, Grunert A, Bachem MG. Lipopolysaccharide-activated macrophages stimulate the synthesis of collagen type I and C-fibronectin in cultured pancreatic stellate cells. Am J Pathol. 1999;155(5):1749–58. doi: 10.1016/S0002-9440(10)65490-9.
    1. Wong L, Hutson PR, Bushman W. Prostatic inflammation induces fibrosis in a mouse model of chronic bacterial infection. PLoS One. 2014;9(6):e100770. doi: 10.1371/journal.pone.0100770.
    1. McCormack VA, dos Santos SI. Breast density and parenchymal patterns as markers of breast cancer risk: a meta-analysis. Cancer Epidemiol Biomarkers Prev. 2006;15(6):1159–69. doi: 10.1158/1055-9965.EPI-06-0034.
    1. Boyd NF, Martin LJ, Stone J, Greenberg C, Minkin S, Yaffe MJ. Mammographic densities as a marker of human breast cancer risk and their use in chemoprevention. Curr Oncol Rep. 2001;3(4):314–21. doi: 10.1007/s11912-001-0083-7.
    1. Ghilardi G, Biondi ML, La Torre A, Battaglioli L, Scorza R. Breast cancer progression and host polymorphisms in the chemokine system: role of the macrophage chemoattractant protein-1 (MCP-1) -2518 G allele. Clin Chem. 2005;51(2):452–5. doi: 10.1373/clinchem.2004.041657.
    1. Rovin BH, Lu L, Saxena R. A novel polymorphism in the MCP-1 gene regulatory region that influences MCP-1 expression. Biochem Biophys Res Commun. 1999;259(2):344–8. doi: 10.1006/bbrc.1999.0796.
    1. Zafiropoulos A, Crikas N, Passam AM, Spandidos DA. Significant involvement of CCR2-64I and CXCL12-3a in the development of sporadic breast cancer. J Med Genet. 2004;41(5):e59. doi: 10.1136/jmg.2003.013649.
    1. Provenzano PP, Inman DR, Eliceiri KW, Knittel JG, Yan L, Rueden CT, White JG, Keely PJ. Collagen density promotes mammary tumor initiation and progression. BMC Med. 2008;6:11. doi: 10.1186/1741-7015-6-11.
    1. Ghosh K, Brandt KR, Reynolds C, Scott CG, Pankratz VS, Riehle DL, Lingle WL, Odogwu T, Radisky DC, Visscher DW, et al. Tissue composition of mammographically dense and non-dense breast tissue. Breast Cancer Res Treat. 2012;131(1):267–75. doi: 10.1007/s10549-011-1727-4.
    1. Guo YP, Martin LJ, Hanna W, Banerjee D, Miller N, Fishell E, Khokha R, Boyd NF. Growth factors and stromal matrix proteins associated with mammographic densities. Cancer Epidemiol Biomarkers Prev. 2001;10(3):243–8.
    1. Chew GL, Huo CW, Huang D, Hill P, Cawson J, Frazer H, Hopper JL, Haviv I, Henderson MA, Britt K, et al. Increased COX-2 expression in epithelial and stromal cells of high mammographic density tissues and in a xenograft model of mammographic density. Breast Cancer Res Treat. 2015;153(1):89–99. doi: 10.1007/s10549-015-3520-2.
    1. McConnell JC, O'Connell OV, Brennan K, Weiping L, Howe M, Joseph L, Knight D, O'Cualain R, Lim Y, Leek A, et al. Increased peri-ductal collagen micro-organization may contribute to raised mammographic density. Breast Cancer Res. 2016;18(1):5. doi: 10.1186/s13058-015-0664-2.
    1. Zhao YS, Zhu S, Li XW, Wang F, Hu FL, Li DD, Zhang WC, Li X. Association between NSAIDs use and breast cancer risk: a systematic review and meta-analysis. Breast Cancer Res Treat. 2009;117(1):141–50. doi: 10.1007/s10549-008-0228-6.
    1. Hugo HJ, Saunders C, Ramsay RG, Thompson EW. New Insights on COX-2 in Chronic Inflammation Driving Breast Cancer Growth and Metastasis. J Mammary Gland Biol Neoplasia. 2015;20(3–4):109–19. doi: 10.1007/s10911-015-9333-4.
    1. Esbona K, Inman D, Saha S, Jeffery J, Schedin P, Wilke L, Keely P. COX-2 modulates mammary tumor progression in response to collagen density. Breast Cancer Res. 2016;18(1):35. doi: 10.1186/s13058-016-0695-3.
    1. Stone J, Willenberg L, Apicella C, Treloar S, Hopper J. The association between mammographic density measures and aspirin or other NSAID use. Breast Cancer Res Treat. 2012;132(1):259–66. doi: 10.1007/s10549-011-1834-2.
    1. Terry MB, Buist DS, Trentham-Dietz A, James-Todd TM, Liao Y. Nonsteroidal anti-inflammatory drugs and change in mammographic density: a cohort study using pharmacy records on over 29,000 postmenopausal women. Cancer Epidemiol Biomarkers Prev. 2008;17(5):1088–95. doi: 10.1158/1055-9965.EPI-07-2836.

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