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K252a induces anoikis-sensitization with suppression of cellular migration in Epstein-Barr Virus (EBV)—associated nasopharyngeal carcinoma cells

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Summary

Recent studies revealed an unexpected role of the neurotrophin receptor pathway, BDNF/TrkB signaling, in cancer metastasis and anoikis (i.e. detachment-induced cell death). Survival of cancer cells in detached state (known as anoikis-resistance) is known to be pre-requisite for metastasis. Nasopharyngeal carcinoma (NPC), an endemic head and neck cancer in Southeast Asia, is highly invasive, metastatic, and etiologically associated with Epstein-Barr virus (EBV, an oncovirus) infection. Mechanistic studies on the invasive/metastatic nature of NPC can facilitate the development of anti-metastatic therapy in NPC. Thus far, the role of BDNF/TrkB signaling in virus-associated human cancer is unclear. Here, using multiple cell line models of NPC with EBV-association (HONE-1-EBV, HK1-LMP1 and C666-1), we investigated the potential involvement of BDNF/TrkB signaling in cellular migration and anoikis-resistant characteristics of NPC. We found that all three EBV-associated NPC cell lines tested were intrinsically anoikis-resistant (i.e. survived in detached state) and expressed both BDNF and TrkB. BDNF stimulation induced cellular migration, but not proliferation of these cells. Further, we examined if pharmacologic targeting of anoikis-resistance of NPC cells can be achievable by a proof-of-concept Trk inhibitor, K252a, in these EBV-associated NPC models. Our results demonstrated that K252a, was able to attenuate BDNF-induced migration and proliferation of NPC cells. More importantly, we demonstrated for the first time that K252a harbored potent anoikis-sensitization activity (i.e. sensitizing cancer cells to detachment-induced cell death) against EBV-associated human cancer cells, namely NPC cells. This proof-of-concept study demonstrated that K252a, a Trk inhibitor, can potentially be used as an anoikis-sensitizing agent in NPC.

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Abbreviations

NPC:

Nasopharyngeal Carcinoma

EBV:

Epstein-Barr virus

LMP1:

Latent Membrane Protein 1

BDNF:

Brain-derived neurotrophic factor

TrkB:

Tropomyosin-related kinase B

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Acknowledgement

This study was supported by Direct Grant for Research, The Chinese University of Hong Kong (2008.1.101 to YKN). Result of this study was presented in part in the poster session at the American Association of Cancer Research (AACR) annual meeting, Colorado, USA, 2009.

Conflict of Interest Statement

All authors have no financial/commercial conflicts of interest regarding the study.

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Authors and Affiliations

  1. Department of Surgery, The Chinese University of Hong Kong, Hong Kong, SAR, China

    Yuen-Keng Ng, Maggie Pui Chun Kwan & Paul Bo San Lai

  2. Department of Clinical Oncology, State Key Laboratory of Oncology in South China, Sir YK Pao Center for Cancer, Hong Kong Cancer Institute and Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong, SAR, China

    Elaine Yue Ling Wong, Cecilia Pik Yuk Lau, Jessica Pui Lan Chan, Sze Chuen Cesar Wong, Andrew Sai-Kit Chan, Anthony Tak Cheung Chan & Vivian Wai Yan Lui

  3. Department of Anatomy, University of Hong Kong, Hong Kong, SAR, China

    Sai-Wah Tsao & Chi-Man Tsang

  4. Department of Clinical Oncology, Sir YK Pao Center for Cancer, Prince of Wales Hospital, Room 302, Shatin, Hong Kong, SAR, China

    Vivian Wai Yan Lui

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  1. Yuen-Keng Ng
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  2. Elaine Yue Ling Wong
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Correspondence to Vivian Wai Yan Lui.

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Supplementary Fig. 1

Exogenous BDNF induced TrkB expression in C666-1. C666-1 cells were treated with either serum free medium with vehicle (H2O) or BDNF (50 ng/ml) for 48 h. The expression level of TrkB was determined by Western blotting. Similar results were obtained in 3 independent experiments. (JPEG 9 kb)

Supplementary Fig. 2

K252a inhibited TrkB phosphorylation in an EBV-associated NPC cell line with high expression level of TrkB. HONE-1-EBV cells were treated with either vehicle (DMSO), 300 nM or 3 μM of K252a for 8 h. Cell lysates were then subjected to immunoprecipitation with TrkB antibody. Expression of phosphorylated TrkB was then detected by immunoblotting with anti-phosphorylated antibody (PY20: BD Transduction Lab, Lexington, KY). Similar results were observed in 3 independent experiments. (JPEG 13 kb)

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Ng, YK., Wong, E.Y.L., Lau, C.P.Y. et al. K252a induces anoikis-sensitization with suppression of cellular migration in Epstein-Barr Virus (EBV)—associated nasopharyngeal carcinoma cells. Invest New Drugs 30, 48–58 (2012). https://doi.org/10.1007/s10637-010-9513-4

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