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GLI-mediated Keratin 17 expression promotes tumor cell growth through the anti-apoptotic function in oral squamous cell carcinomas

  • Original Article – Cancer Research
  • Published:
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Abstract

Purpose

Keratin 17 (KRT17) has been suggested as a potential diagnostic marker of squamous cell carcinoma including oral squamous cell carcinoma (OSCC). The current study was conducted to clarify the function of KRT17 and its expression mechanism in OSCC.

Methods

Immunohistochemical analyses were carried out to examine the expression of KRT17, GLI family zinc finger (GLI)-1, GLI-2, or cleaved caspase-3 in OSCCs. The expression of KRT17, GLI-1, or GLI-2 was investigated among OSCC cell lines, and the effects of loss-of-function of KRT17 or GLI, using siRNA or inhibitor, on the cell growth of the OSCC cell line HSC-2 particularly with respect to apoptosis were examined.

Results

Immunohistochemical analyses of tissue specimens obtained from 78 OSCC patients revealed that KRT17 was not observed in non-tumor regions but was strongly expressed at high frequencies in tumor regions. Knockdown of KRT17 increased the number of cleaved caspase-3-positive cells, leading to the reduction of cell number. Loss-of-function of GLI-1 or GLI-2 also increased the cell numbers of apoptotic cells positive for staining of Annexin-V and propidium iodide (PI) and the terminal deoxynucleotidyl transferase dUTP-biotin nick-end labeling (TUNEL) method, and induced DNA fragmentation. This inhibitory effect on cell growth was partially rescued by exogenous KRT17 expression. In the KRT17-positive regions in OSCCs, GLI-1 or GLI-2 was frequently detected, and the number of cells with cleaved caspase-3 positive was decreased.

Conclusions

KRT17 promotes tumor cell growth, at least partially, through its anti-apoptotic effect as a result of the KRT17 overexpression by GLIs in OSCC.

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Acknowledgements

The authors thank Dr. R. Kitamura for valuable help regarding statistical analyses and Dr. H. Fujiwara for assistance with the cell death assay and DNA fragmentation.

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Author notes

    Authors and Affiliations

    1. Laboratory of Oral Pathology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

      Yurie Mikami, Shinsuke Fujii, Kengo Nagata, Hiroko Wada, Kana Hasegawa, Misaki Abe, Reiko U. Yoshimoto & Tamotsu Kiyoshima

    2. Section of Maxillofacial Oncology, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

      Yurie Mikami, Shintaro Kawano & Seiji Nakamura

    3. Section of Oral and Maxillofacial Surgery, Division of Maxillofacial Diagnostic and Surgical Sciences, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

      Misaki Abe

    4. Department of Periodontology, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

      Reiko U. Yoshimoto

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    1. Yurie Mikami
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    2. Shinsuke Fujii
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    Correspondence to Tamotsu Kiyoshima.

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    The authors declare that they have no conflicts of interest.

    Ethical approval

    Ethical approval for this study was obtained from the institutional review board of Kyushu University Hospital.

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    Written informed consent was obtained from individual participants.

    Funding

    This work was supported by Grants-in-Aid for Scientific Research to K. T. (No. 15K15695), S. F. (No. 16K11501), and H. W. (No. 26462788) from the Ministry of Education, Science, and Culture of Japan.

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    Y. Mikami and S. Fujii have contributed equally to this work.

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    Mikami, Y., Fujii, S., Nagata, K. et al. GLI-mediated Keratin 17 expression promotes tumor cell growth through the anti-apoptotic function in oral squamous cell carcinomas. J Cancer Res Clin Oncol 143, 1381–1393 (2017). https://doi.org/10.1007/s00432-017-2398-2

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    • DOI: https://doi.org/10.1007/s00432-017-2398-2

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