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Clinical significance of CYLD downregulation in breast cancer

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Abstract

Cylindromatosis (CYLD) is a tumor suppressor gene that is mutated in familial cylindromatosis, a rare autosomal dominant disorder associated with numerous benign skin adnexal tumors. CYLD is now known to regulate various signaling pathways, including transforming growth factor-β signaling, Wnt/β-catenin signaling, and NF-κB signaling by deubiquitinating upstream regulatory factors. Downregulation of CYLD has been reported in several malignancies; however, the clinical significance of CYLD expression in many malignancies, including breast cancer, remains to be elucidated. This study investigated the clinical significance of CYLD in breast cancer and its roles in tumor progression. We evaluated CYLD expression in matched normal breast tissue samples and tumor breast tissue samples from 26 patients with breast cancer and in a series of breast cancer cell lines. In addition, by means of immunohistochemistry, we investigated CYLD protein expression and its clinical significance in 244 breast cancer cases. We also analyzed the effects of CYLD repression or overexpression on breast cancer cell viability, cell migration, and NF-κB activity with or without receptor activator of NF-κB ligand (RANKL) stimulation. Breast cancer tissues demonstrated significantly reduced CYLD mRNA expression compared with normal breast tissues. Downregulation of CYLD promoted cell survival and migratory activities through NF-κB activation, whereas CYLD overexpression inhibited those activities in MDA-MB-231 cells. As an important finding, CYLD overexpression also inhibited RANKL-induced NF-κB activation. Our immunohistochemical analysis revealed that reduced CYLD protein expression was significantly correlated with estrogen receptor negativity, high Ki-67 index, high nuclear grade, decreased disease-free survival, and reduced breast cancer-specific survival in primary breast cancer. Moreover, reduced CYLD expression was an independent factor for poor prognosis in breast cancer. CYLD downregulation may promote breast cancer metastasis via NF-κB activation, including RANKL signaling.

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Abbreviations

CYLD:

Cylindromatosis

RANKL:

NF-κB ligand

HER2:

Human epidermal growth factor receptor 2

ER:

Estrogen receptor

PgR:

Progesterone receptor

HMEC:

Human mammary epithelial cells

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Acknowledgments

The authors thank Dr. Seiji Okada and Dr. Manabu Taura for supplying pGL4b and phRG-TK vectors; Y. Ogasawara, K. Kai, M. Imamoto, E. Nakamura, and H. Katsura for excellent technical support; Dr. K. Iyama, Dr. Y. Honda, and Dr. T. Asato for their excellent pathological diagnosis. The authors’ work was supported by a Grand-in-Aid for Scientific Research (A) 24249036 (Y. Ando), by Grand-in-Aid for Young Scientists (B) 23790091 (H. Jono) and (B) 24792238 (S. Shinriki) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

Conflict of interest

The authors disclosed no conflicts of interest.

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Authors and Affiliations

  1. Department of Breast and Endocrine Surgery, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

    Mitsuhiro Hayashi, Aiko Sueta, Mai Tomiguchi, Saori Fujiwara, Mutsuko Yamamoto-Ibusuki, Kei-ichi Murakami, Yutaka Yamamoto & Hirotaka Iwase

  2. Department of Pharmacy, Kumamoto University Hospital, Kumamoto, Japan

    Hirofumi Jono

  3. Department of Laboratory Medicine, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto, Kumamoto, 860-8556, Japan

    Satoru Shinriki

  4. Department of Oral and Maxillofacial Surgery, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

    Takuya Nakamura

  5. Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

    Jianying Guo, Satoshi Yamashita & Yukio Ando

  6. Center for Inflammation, Immunity and Infection and Department of Biology, Georgia State University, Atlanta, GA, USA

    Jian-Dong Li

Authors
  1. Mitsuhiro Hayashi
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  2. Hirofumi Jono
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  3. Satoru Shinriki
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  4. Takuya Nakamura
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  9. Mutsuko Yamamoto-Ibusuki
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  11. Satoshi Yamashita
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  12. Yutaka Yamamoto
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  13. Jian-Dong Li
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  14. Hirotaka Iwase
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  15. Yukio Ando
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Correspondence to Satoru Shinriki.

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Hayashi, M., Jono, H., Shinriki, S. et al. Clinical significance of CYLD downregulation in breast cancer. Breast Cancer Res Treat 143, 447–457 (2014). https://doi.org/10.1007/s10549-013-2824-3

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