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The estrogen receptor beta agonist liquiritigenin enhances the inhibitory effects of the cholesterol biosynthesis inhibitor RO 48-8071 on hormone-dependent breast-cancer growth

  • Preclinical study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Purpose

Most hormone-dependent human breast cancers develop resistance to anti-hormone therapy over time. Our goal was to identify novel treatment strategies to avoid this drug resistance and thereby control hormone-dependent breast cancer.

Methods

Sulforhodamine B assays were used to measure viability of cultured human breast-cancer cells. BT-474 cell tumor xenografts in nude mice were used to evaluate tumor growth. Immunohistochemistry was used to assess estrogen-receptor and angiogenesis-marker expression, as well as apoptosis, in tumor-xenograft tissues.

Results

MCF-7 and BT-474 breast-cancer cells treated with either RO 48–8071 <[4′-[6-(Allylmethylamino)hexyloxy]-4-bromo-2′-fluorobenzophenone fumarate] [RO]; a small-molecule inhibitor of oxidosqualene cyclase, a key enzyme in cholesterol biosynthesis> or liquiritigenin [LQ; an estrogen receptor (ER) β agonist] exhibited significantly reduced viability in vitro. RO + LQ treatment further significantly reduced cell viability. Administration of RO, LQ, or RO + LQ significantly inhibited growth of BT-474 tumor xenografts in vivo. RO, LQ, or RO + LQ reduced ERα but induced ER β expression in tumor xenografts. Both compounds significantly reduced angiogenesis-marker expression and increased apoptosis in tumor xenografts; use of RO + LQ significantly enhanced the effects observed with a single agent.

Conclusion

The ERβ ligand LQ significantly enhanced the inhibition of breast-cancer cell viability and tumor-xenograft growth by RO. The anti-tumor properties of RO may in part be due to an off-target effect that reduces ERα and increases ERβ, the latter of which can then interact with LQ to promote anti-proliferative effects. The RO + LQ combination may have value when considering novel treatment strategies for hormone-dependent breast cancer.

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Fig. 1
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Fig. 6

taken from 4 to 5 animals and 26–30 areas were quantitated for VEGF, and 26–30 images were quantitated for CD31. Values represent means ± SEM. Insets for representative micrographs represent negative controls. Bars represent 50 μm. *Significantly different from controls; **Significantly different from all other groups (P < 0.05 using ANOVA)

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Data availability

Data will be available on request.

Abbreviations

ANOVA:

Analysis of variance

DCC:

Dextran-coated charcoal

ER:

Estrogen receptor

FBS:

Fetal bovine serum

ip:

Intraperitoneal

iv:

Intravenous

LQ:

Liquiritigenin

OSC:

Oxidosqualene cyclase

PBS:

Phosphate-buffered saline

RO:

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

sc:

Subcutaneous

SEM:

Standard error of the mean

SRB:

Sulforhodamine B

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

VEGF:

Vascular endothelial growth factor

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Acknowledgements

Supported in part by a Department of Defense Breast Cancer Program Grant W81XWH-12-1-0191, and by a Faculty Research Grant from the University of Missouri, Columbia. SMH is the Zalk Missouri Professor of Tumor Angiogenesis.

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Authors and Affiliations

  1. Dalton Cardiovascular Research Center, University of Missouri, 134 Research Park Drive, Columbia, MO, 65211, USA

    Yayun Liang & Salman M. Hyder

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

    Yayun Liang & Salman M. Hyder

  3. IDEXX BioAnalytics, Columbia, MO, 65202, USA

    Cynthia Besch-Williford

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  1. Yayun Liang
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Correspondence to Salman M. Hyder.

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All animal studies were approved by ACUC at the University of Missouri.

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Liang, Y., Besch-Williford, C. & Hyder, S.M. The estrogen receptor beta agonist liquiritigenin enhances the inhibitory effects of the cholesterol biosynthesis inhibitor RO 48-8071 on hormone-dependent breast-cancer growth. Breast Cancer Res Treat 192, 53–63 (2022). https://doi.org/10.1007/s10549-021-06487-y

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