Abstract
FK506-binding protein 51 (FKBP51) is an immunophilin with isomerase activity, which performs important biological functions in the cell. It has recently been involved in the apoptosis resistance of malignant melanoma. The aim of this study was to investigate the possible role of FKBP51 in the control of response to ionizing radiation (Rx) in malignant melanoma. FKBP51-silenced cells showed reduced clonogenic potential after irradiation compared with non-silenced cells. After Rx, we observed apoptosis in FKBP51-silenced cells and autophagy in non-silenced cells. The FKBP51-controlled radioresistance mechanism involves NF-κB. FKBP51 was required for the activation of Rx-induced NF-κB, which in turn inhibited apoptosis by stimulating X-linked inhibitor of apoptosis protein and promoting authophagy-mediated Bax degradation. Using a tumor–xenograft mouse model, the in vivo pretreatment of tumors with FKBP51-siRNA provoked massive apoptosis after irradiation. Immunohistochemical analysis of 10 normal skin samples and 80 malignant cutaneous melanomas showed that FKBP51 is a marker of melanocyte malignancy, correlating with vertical growth phase and lesion thickness. Finally, we provide evidence that FKBP51 targeting radiosensitizes cancer stem/initiating cells. In conclusion, our study identifies a possible molecular target for radiosensitizing therapeutic strategies against malignant melanoma.
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Abbreviations
- ABCG2:
-
ATP-binding cassette subfamily G member 2
- Becn-1:
-
Beclin-1
- CMM:
-
cutaneous malignant melanoma
- FKBP51:
-
FK506 binding protein 51
- IκB:
-
inhibitor of κ-light-chain-enhancer of activated B cells
- IKK:
-
IκB kinase complex
- LC3:
-
light chain 3
- MFI:
-
mean fluorescence intensity
- NF-κB:
-
nuclear factor of κ-light-chain-enhancer of activated B cells
- NS:
-
non-silencing
- Rx:
-
ionizing radiation
- siRNA:
-
short interfering RNA
- xIAP:
-
X-linked inhibitor of apoptosis protein
- zVAD:
-
z-Valine–Alanine–Aspartic acid
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Acknowledgements
The authors are grateful to Professor Eric Baehrecke (University of Massachusetts Medical School, Worcester, MA, USA) for advice and helpful discussion, Dr. Raffaele Liuzzi (Institute of Biostructure and Bio-Imaging National Research Council, Naples, Italy) for contribution to statistical analysis, Professor Luigi Del Vecchio (CEINGE, Naples, Italy) for cell sorting, the Electron Microscopy Unit at the Institute of Biotechnology, University of Helsinki for access to equipment and technical help. It is stated that no conflict of interest exists. The work is supported by funds from the Italian Association for Cancer Research (AIRC) and MIUR (Ministero Istruzione, Università e Ricerca).
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Romano, S., D'Angelillo, A., Pacelli, R. et al. Role of FK506-binding protein 51 in the control of apoptosis of irradiated melanoma cells. Cell Death Differ 17, 145–157 (2010). https://doi.org/10.1038/cdd.2009.115
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DOI: https://doi.org/10.1038/cdd.2009.115
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