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Cholesterol biosynthesis inhibitors as potent novel anti-cancer agents: suppression of hormone-dependent breast cancer by the oxidosqualene cyclase inhibitor RO 48-8071

  • Preclinical study
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Abstract

In most human breast cancers, tumor cell proliferation is estrogen dependent. Although hormone-responsive tumors initially respond to anti-estrogen therapies, most of them eventually develop resistance. Our goal was to identify alternative targets that might be regulated to control breast cancer progression. Sulforhodamine B assay was used to measure the viability of cultured human breast cancer cell lines exposed to various inhibitors. Protein expression in whole-cell extracts was determined by Western blotting. BT-474 tumor xenografts in nude mice were used for in vivo studies of tumor progression. RO 48-8071 ([4′-[6-(Allylmethylamino)hexyloxy]-4-bromo-2′-fluorobenzophenone fumarate]; RO), a small-molecule inhibitor of oxidosqualene cyclase (OSC, a key enzyme in cholesterol biosynthesis), potently reduced breast cancer cell viability. In vitro exposure of estrogen receptor (ER)-positive human breast cancer cells to pharmacological levels of RO or a dose close to the IC50 for OSC (nM) reduced cell viability. Administration of RO to mice with BT-474 tumor xenografts prevented tumor growth, with no apparent toxicity. RO degraded ERα while concomitantly inducing the anti-proliferative protein ERβ. Two other cholesterol-lowering drugs, Fluvastatin and Simvastatin, were less effective in reducing breast cancer cell viability and were found not to induce ERβ. ERβ inhibition or knockdown prevented RO-dependent loss of cell viability. Importantly, RO had no effect on the viability of normal human mammary cells. RO is a potent inhibitor of hormone-dependent human breast cancer cell proliferation. The anti-tumor properties of RO appear to be in part due to an off-target effect that increases the ratio of ERβ/ERα in breast cancer cells.

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Abbreviations

E:

Estrogen

ER:

Estrogen receptor

PR:

Progesterone receptor

OSC:

Oxidosqualene cyclase

RO:

RO 48-8071 ([4′-[6-(Allylmethylamino)hexyloxy]-4-bromo-2′-fluorobenzophenone fumarate])

FBS:

Fetal bovine serum

SRB:

Sulforhodamine B

PI:

Propidium iodide

sc:

Subcutaneous

iv:

Intravenous

PBS:

Phosphate-buffered saline

TBS-T:

Tris-buffered saline containing 0.1 % Tween 20

ANOVA:

Analysis of variance

SE:

Standard error

DPN:

2,3-bis(4-Hydroxyphenyl)-propionitrile

PHTPP:

4-[2-Phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol

FACS:

Fluorescence-activated cell sorting

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Acknowledgments

Supported by a Department of Defense Breast Cancer Program Grant W81XWH-12-1-0191, the National Institutes of Health Grant R21 GM088517, and by a Faculty Research Grant from the University of Missouri, Columbia. SMH is the Zalk Missouri Professor of Tumor Angiogenesis. Funds to purchase the nanodrop instrument were provided through the generosity of numerous donors to the Ellis Fischel Cancer Center.

Conflict of interest

JDA is an employee of F. Hoffmann-La Roche AG. All other authors declare that they have no conflict of interest.

Ethical standards

The experiments comply with the current laws of the country in which they were performed (USA).

Author information

Authors and Affiliations

  1. Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO, 65211, USA

    Yayun Liang, Benford Mafuvadze, Matthew T. Cook, Xiaoqin Zou & Salman M. Hyder

  2. Department of Biomedical Sciences, University of Missouri, Columbia, MO, 65211, USA

    Yayun Liang, Benford Mafuvadze, Matthew T. Cook & Salman M. Hyder

  3. IDDEX RADIL, Columbia, MO, 65202, USA

    Cynthia Besch-Williford

  4. Small Molecule Research, Medicinal Chemistry, Pharma Research & Early Development (pRED), F. Hoffmann-La Roche AG, Grenzacherstrasse 124, 4070, Basel, Switzerland

    Johannes D. Aebi

  5. Department of Physics and Astronomy, Department of Biochemistry, and Informatics Institute, University of Missouri, Columbia, MO, 65211, USA

    Xiaoqin Zou

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  1. Yayun Liang
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  2. Cynthia Besch-Williford
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Correspondence to Salman M. Hyder.

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Liang, Y., Besch-Williford, C., Aebi, J.D. et al. Cholesterol biosynthesis inhibitors as potent novel anti-cancer agents: suppression of hormone-dependent breast cancer by the oxidosqualene cyclase inhibitor RO 48-8071. Breast Cancer Res Treat 146, 51–62 (2014). https://doi.org/10.1007/s10549-014-2996-5

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  • DOI: https://doi.org/10.1007/s10549-014-2996-5

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