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. 2017 Oct 11;8:864. doi: 10.1038/s41467-017-00910-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — 27HC is a biochemical mediator of the effects of obesity and elevated cholesterol on breast cancer metastasis. a A high-cholesterol diet (HCD) increases spontaneous metastasis of murine ER-negative Met1 cells (N = 10 each). b Mice fed a high-cholesterol diet (HCD) exhibit increased lung colonization of i.v. grafted Met1 mammary tumors (CD N = 7, HCD N = 8). c A high-fat diet (HFD) increases lung colonization of murine E0771 i.v. grafts in APOE3 mice (i.v. graft of cells). Pretreatment for 7 days prior to the graft and continued treatment throughout the study with daily placebo or atorvastatin (statin, 40 mg/kg) by oral gavage reduced metastatic colonization. Representative images of iRFP-expressing E0771 colonies (depicted as black) within the lungs are presented above quantified data (control diet (CD) placebo N = 11, statin N = 7, HFD placebo N = 8, statin N = 7, scale bar indicates 5 mm). d Chronic administration of exogenous 27-hydroxycholesterol (27HC) increases spontaneous metastasis to the lung as measured by relative fluorescence (inset) without impacting growth rate of the primary tumor (N = 3). e Metastatic effects of a HCD in MMTV-PyMT mice requires conversion to 27HC by CYP27A1 (wild-type CD N = 8, HCD N = 14, CYP27A1^−/− CD N = 3, HCD N = 9). Quantitative RT-PCR for PyMT transcript within the lung is presented in the graph and used to quantify metastasis since its expression should be restricted to mammary epithelial cells. f Chronic treatment with a small molecule inhibitor of CYP27A1, GW273297X (GW297X) decreases colonization of lung tissue from Met1 i.v. grafts (N = 4 each). g Lung colonization from E0771 i.v. grafts in APOE3 mice is increased by a HFD, but attenuated in mice chronically treated with GW297X (CD placebo N = 5, GW297X N = 3, HFD placebo N = 7, GW297X N = 6). Results are depicted as mean +/− SEM. Lines and asterisks denote statistical differences between groups (p < 0.05). [a, b, d, f: Unpaired two-tailed student’s t-test. c, e, g: One-way ANOVA followed by a Student Newman-Keuls multiple comparison test]