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TRIP12 as a mediator of human papillomavirus/p16-related radiation enhancement effects

Oncogene volume 36, pages 820–828 (2017)Cite this article

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Abstract

Patients with human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) have better responses to radiotherapy and higher overall survival rates than do patients with HPV-negative HNSCC, but the mechanisms underlying this phenomenon are unknown. p16 is used as a surrogate marker for HPV infection. Our goal was to examine the role of p16 in HPV-related favorable treatment outcomes and to investigate the mechanisms by which p16 may regulate radiosensitivity. HNSCC cells and xenografts (HPV/p16-positive and -negative) were used. p16-overexpressing and small hairpin RNA-knockdown cells were generated, and the effect of p16 on radiosensitivity was determined by clonogenic cell survival and tumor growth delay assays. DNA double-strand breaks (DSBs) were assessed by immunofluorescence analysis of 53BP1 foci; DSB levels were determined by neutral comet assay; western blotting was used to evaluate protein changes; changes in protein half-life were tested with a cycloheximide assay; gene expression was examined by real-time polymerase chain reaction; and data from The Cancer Genome Atlas HNSCC project were analyzed. p16 overexpression led to downregulation of TRIP12, which in turn led to increased RNF168 levels, repressed DNA damage repair (DDR), increased 53BP1 foci and enhanced radioresponsiveness. Inhibition of TRIP12 expression further led to radiosensitization, and overexpression of TRIP12 was associated with poor survival in patients with HPV-positive HNSCC. These findings reveal that p16 participates in radiosensitization through influencing DDR and support the rationale of blocking TRIP12 to improve radiotherapy outcomes.

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Acknowledgements

We thank Dr Jeffery Myers and Dr Peter J F Snijders for kindly sharing their cell lines. This work was supported by grants R01CA168485 and R01CA181029 from the National Cancer Institute (REM and KKA) and (LM), respectively, the Gilbert H Fletcher Chair (KKA), Cancer Center Support Grant (P30 NCI CA016672), Individual Investigator Research Award RP150293 from the Cancer Prevention Research Institute of Texas (HDS), an MD Anderson Cancer Head and Cancer SPORE Career Development Award (HDS) and a Center for Radiation Oncology Research Development Award (HDS). The content is solely the responsibility of the authors and does not represent the official views of any grant-awarding agency.

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  1. L Wang and P Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    L Wang, P Zhang, D P Molkentine, C Chen, J M Molkentine, H Piao, U Raju, J Zhang, D R Valdecanas, H D Thames, J Chen, L Ma, K A Mason, K-K Ang & R E Meyn

  2. Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China

    H Piao

  3. Department of Radiation Physics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    R C Tailor

  4. Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    T A Buchholz, K-K Ang & H D Skinner

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Wang, L., Zhang, P., Molkentine, D. et al. TRIP12 as a mediator of human papillomavirus/p16-related radiation enhancement effects. Oncogene 36, 820–828 (2017). https://doi.org/10.1038/onc.2016.250

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