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Identification of estrogen-responsive genes involved in breast cancer metastases to the bone

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Abstract

Bone metastasis is the most common metastasis in breast cancer patients. Clinical observations propose strong association between estrogen receptor (ER)-positive tumors and the development of bone metastases. We hypothesized of biologically diverse sets of hormone-dependent tumors predisposed to bone metastases and of possible role of ER-signaling pathways in the development and progression of bone metastases. We developed a novel in vitro estrogen (E2)-responsive model system, in which breast cancer cells and bone cells express high levels of either ERα or ERβ. Using co-culture approach and gene array technology we identified E2-responsive genes involved in the interaction between cancer cells and bone cells. We detected 13 genes that were altered solely by ERα and 11 genes that were regulated solely by ERβ in cancer cells. Only 5 genes were modified by both ERα and ERβ. Interestingly, the majority of genes in bone cells were altered through ERβ. Two genes, namely MacMarcks and Muc-1, whose changes in expressions in cancer cells in response to E2 were highly significant, were selected for immunohistochemical analysis using tissue microarrays of 59 infiltrating ductal carcinomas. Our results indicated that both MacMarcks and Muc-1 were expressed at high frequency in ER-positive tumors. The correlation between ERα- and ERβ-status of hormone-dependent tumors with combined expression of these two markers might suggest a more aggressive tumor phenotype associated with bone metastases. Further analysis of tissues with clinicopathological characteristics and known bone metastatic disease will indicate potential prognostic values of these and other markers in the development of bone metastases in a subgroup of “bad” hormone-dependent breast cancer.

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Abbreviations

ERα:

Estrogen receptor alpha

ERβ:

Estrogen receptor beta

E2:

17-β estradiol

F:

Fulvestrant

Dox:

Doxycycline

MDA-ERα:

MDA-MB-231 cells stably transfected with ERα

MDA-ERβ:

MDA-MB-231 cells stably transfected with ERβ

U2OS:

Osteosarcoma cells

TMA:

Tissue microarrays

ERE:

Estrogen response element

RT-PCR:

Reverse transcriptase polymerase chain reaction

MEM:

Minimum essential medium

DMEM/F12:

Dulbecco’s Modified Eagle’s medium

IHC:

Immunohistochemical

ECM:

Extracellular matrix

MMP:

Matrix metalloproteinase

TGFβ:

Transforming growth factor beta

FGF:

Fibroblast growth factor

OPN:

Osteopontin

MacMarcks:

Myristoylated alanine-rich C kinase substrate

Muc-1:

Mucin 1

PKC:

Protein kinase C

MAP:

Mitogen activated protein

RANK:

Receptor activator of NF-κB

RANKL:

Receptor activator of NF-κB ligand

CFS:

Colony-stimulating factor

ErbB-2:

Erythroblastic leukemia viral oncogene homolog 2

vs:

Versus

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Acknowledgments

This work was supported by a grant from Susan G Komen Breast Cancer Foundation (BCTR0504398) to ASL. We thank Dr. T. Spelsberg (Mayo Clinic and Foundation, Rochester, MN) for generous gift of U2OS-ER (α and β) cells and Dr. D. Tonetti (University of Illinois at Chicago, Chicago, IL) for MDA-ERβ cells. We are also grateful to Dr. J-A Gustafsson (Karolinska Institute, Sweden) for providing ERβ antibodies for Western blots.

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Feinberg School of Medicine, Northwestern University, 645 N. Michigan Avenue, S910, Chicago, IL, 60611, USA

    Jun Wang, Jennifer Jarrett, Robert L. Satcher Jr. & Anait S. Levenson

  2. Department of Preventive Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL, 60611, USA

    Chiang-Ching Huang

  3. The Robert H. Lurie Comprehensive Cancer Center, The Feinberg School of Medicine, Northwestern University, Chicago, IL, 60611, USA

    Robert L. Satcher Jr. & Anait S. Levenson

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  1. Jun Wang
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  2. Jennifer Jarrett
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  5. Anait S. Levenson
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Correspondence to Anait S. Levenson.

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Wang, J., Jarrett, J., Huang, CC. et al. Identification of estrogen-responsive genes involved in breast cancer metastases to the bone. Clin Exp Metastasis 24, 411–422 (2007). https://doi.org/10.1007/s10585-007-9078-6

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