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Up-regulation of cathepsin S expression by HSP90 and 5-HT7 receptor-dependent serotonin signaling correlates with triple negativity of human breast cancer

  • Preclinical study
  • Published:
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Abstract

Purpose

Cathepsin S (CTSS) is expressed in a variety of cancers and stimulates tumor progression. However, the regulatory mechanism and role of CTSS in breast cancer progression are poorly understood. The aim of this study was to examine the relationships between CTSS expression and breast cancer grade and stage, and the signaling molecules involved in CTSS expression.

Methods

Immunohistochemical staining was performed in tissue microarray sections of 1451 human invasive breast cancer samples to determine epithelial (E-CTSS) and stromal CTSS (S-CTSS) expression. Gene and protein expression levels in human breast cancer cell lines were measured by polymerase chain reaction and western blotting. Small interfering RNA transfection and a Matrigel transwell invasion assay were used to confirm the signaling pathways regulating CTSS expression.

Results

In patient tumor tissue blocks, high grade, late stage, and triple negativity were associated with elevated CTSS protein expression, and expression levels were related to the clinical outcomes of patients with invasive breast cancer. CTSS expression was also higher in triple-negative breast cancer (TNBC) cell lines than in hormone-responsive cells, and CTSS expression patterns matched those of tryptophan hydroxylase 1 (TPH1) and 5-hydroxytryptamine receptor 7 (5-HT7). Treatment of TNBC cells (MDA-MB-231 and HCC-1395) with 5-HT significantly enhanced CTSS protein expression, whereas pharmacological inhibition or knockdown of 5-HT7 significantly inhibited its expression. Correspondingly, cancer cell invasion was increased by 5-HT treatment and suppressed by 5-HT7 knockdown. The expression of CTSS was regulated by PI3K/Akt and Ras/Raf/MAPK signaling pathways, and these signaling pathways were stabilized by HSP90 and enhanced by the 5-HT7 receptor-dependent autocrine effect of 5-HT in TNBC cells.

Conclusion

Our findings suggest CTSS as a candidate target for development of a strategy to inhibit breast cancer invasion, and indicate that HSP90 and 5-HT7 (regulators of CTSS) should be considered as alternative targets for the management of TNBC invasion and metastasis.

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Abbreviations

5-HT:

5-Hydroxytryptamine

5-HT7 :

5-Hydroxytryptamine receptor 7

AP-1:

Activator protein 1

CTSS:

Cathepsin S

E-CTSS:

Epithelial cathepsin S

ER:

Estrogen receptor

ERK:

Extracellular signal-regulated kinase

FBS:

Fetal bovine serum

GA:

Geldanamycin

GFR:

Growth factor receptor

HSP90:

Heat shock protein 90

IKKβ:

Inhibitor kappa-B kinase beta

MAPK:

Mitogen-activated protein kinase

MMP-9:

Matrix metalloproteinase-9

NF-κB:

Nuclear factor-κB

PI3K:

Phosphatidylinositol-3 kinase

PR:

Progesterone receptor

S-CTSS:

Stromal cathepsin S

Sp-1:

Specificity protein-1

TNBC:

Triple-negative breast cancer

TPH1:

Tryptophan hydroxylase 1

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Acknowledgments

This work was supported by a Yeungnam University Research Grant (2012).

Author contributions

JAK conceived and designed the study with YKB. YKB performed immunohistochemical staining and analyses on human breast cancer tissue microarray sections. JG performed experiments with cell lines, and analyzed the data with JAK. All authors participated in the manuscript writing, and have approved the final version of the manuscript.

Author information

Authors and Affiliations

  1. College of Pharmacy, Yeungnam University, Gyeongsan, 38541, Republic of Korea

    Jaya Gautam & Jung-Ae Kim

  2. Department of Pathology, College of Medicine, Yeungnam University, Daegu, 42415, Republic of Korea

    Young Kyung Bae

Authors
  1. Jaya Gautam
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Correspondence to Young Kyung Bae or Jung-Ae Kim.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval

The study on CTSS expression in the tissue microarray sections of 1451 human invasive breast cancer samples was approved by the Institutional Review Board of Yeungnam University Hospital (2015-06-007), and the requirement for informed consent was waived.

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Electronic supplementary material

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Supplementary Fig. S1 Treatment with GA had no effect on MDA-MB-231 cell viability.

Supplementary Fig. S2 GA (an HSP90 inhibitor) blocked the nuclear translocation of NF-κB in MDA-MB-231 cells. Supplementary material 1 (TIFF 3388 kb)

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Gautam, J., Bae, Y.K. & Kim, JA. Up-regulation of cathepsin S expression by HSP90 and 5-HT7 receptor-dependent serotonin signaling correlates with triple negativity of human breast cancer. Breast Cancer Res Treat 161, 29–40 (2017). https://doi.org/10.1007/s10549-016-4027-1

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  • DOI: https://doi.org/10.1007/s10549-016-4027-1

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