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Anti-invasion, anti-proliferation and anoikis-sensitization activities of lapatinib in nasopharyngeal carcinoma cells

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Summary

Nasopharyngeal cancer (NPC) is a highly prevalent and invasive head and neck cancer in Asia. Disease recurrence and distant metastasis account for major NPC deaths. Therefore, more effective therapy is needed. Lapatinib, a dual tyrosine kinase inhibitor (TKI) against both EGFR and HER-2, has been known to exert potent antitumor activity against several cancer models. Given that both EGFR and HER-2 are co-expressed in NPC, we hypothesized that dual targeting of EGFR and HER-2 by this small molecule EGFR/HER-2 TKI would elicit anti-tumor activity in NPC. Using in vitro models of NPC, we demonstrated that lapatinib was able to efficiently inhibit the phosphorylation of both EGFR and HER-2. This was accompanied by significant growth inhibition of NPC cells (with maximal growth inhibition >90%). For the most lapatinib-sensitive cell line (HK1-LMP1, with IC50 ∼ 600 nM), which harbored the highest levels of both EGFR and HER-2, inhibition of cell growth was associated G0/G1 cell cycle arrest, marked PARP cleavage, caspase-3 cleavage, as well as significant downregulation of several important survival proteins (e.g. survivin, Mcl-1 and cyclin D1). NPC cells are intrinsically invasive. We found that lapatinib was able to inhibit cellular invasion of both HK1-LMP1 and HONE-1 cells. Furthermore, our data demonstrated for the first time that lapatinib harbored potent anoikis-sensitization activity (i.e. sensitizing cancer cells to detachment-induced apoptosis) in human cancer cells overexpressing both EGFR and HER-2 (HK1-LMP1 and HK1). Taken together, our findings suggest that lapatinib is a promising anti-cancer agent for NPC with anti-invasion and anoikis-sensitization activities.

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Abbreviations

NPC:

Nasopharyngeal carcinoma

LMP1:

Latent membrane protein 1

EGFR or ErbB1:

Epidermal Growth Factor Receptor

HER-2 or ErbB2:

Human Epidermal growth factor Receptor 2

FISH:

Fluorescence in situ hybridization

TKI:

Tyrosine kinase inhibitor

PARP:

Poly (ADP-ribose) polymerase

Mcl-1:

Induced myeloid leukemia cell differentiation protein

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Acknowledgement

This work was supported by a research grant from GlaxoSmithKline, UK (to T. Mok).

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Authors and Affiliations

  1. Department of Clinical Oncology, Chinese University of Hong Kong, Hong Kong, SAR, Hong Kong

    Vivian Wai Yan Lui, Kenny Ieng Kit Lei, Anthony TC Chan & Tony Shu Kam Mok

  2. Cancer Drug Testing Unit, State Key Laboratory in Oncology in South China, Sir YK Pao Center for Cancer, Department of Clinical Oncology, Hong Kong Cancer Institute, Hong Kong, SAR, Hong Kong

    Vivian Wai Yan Lui, Cecilia Pik Yuk Lau & KaKiu Ho

  3. Department of Anatomical and Cellular Pathology, Chinese University of Hong Kong, Hong Kong, SAR, Hong Kong

    Margaret Heung Ling Ng & Suk Hang Cheng

  4. Department of Anatomy, University of Hong Kong, Pokfulam, Hong Kong

    Sai-Wah Tsao & Chi Man Tsang

  5. Department of Clinical Oncology, Prince of Wales Hospital, Shatin, Hong Kong

    Vivian Wai Yan Lui

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  1. Vivian Wai Yan Lui
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  2. Cecilia Pik Yuk Lau
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  3. KaKiu Ho
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  9. Anthony TC Chan
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  10. Tony Shu Kam Mok
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Correspondence to Vivian Wai Yan Lui.

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Vivian Wai Yan Lui and Cecilia Pik Yuk Lau contributed equally to this work.

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Supplementary Figure 1

Lapatinib inhibited cellular invasion of NPC cells through the Matrigel Invasion chambers. Invaded cells were fixed and stained with Toludine Blue O. Photographs were taken at 200 × magnification under a light microscope. (JPEG 50 kb)

Supplementary Figure 2

High endogenous levels of total and phosphorylated EGFR and HER-2 in HK1 and HK1-LMP1 cells. Total and phosphorylated EGFR were detected in HK1 and HK1-LMP1 cells by Western blotting. Total HER-2 and phosphorylated HER-2 were detected by Western blotting and immunoprecipitation, respectively. Similar results were obtained in three independent experiments. (JPEG 28 kb)

Supplementary Figure 3

Lapatinib induced anoikis in HK1 cells. Lapatinib treatment (600 nM for 48 hrs) markedly reduced the size of multi-cellular spheroids of HK1 cells, which was associated with significant growth inhibition and marked increase in apoptosis activity in detached state (as quantified by Cell Death ELISA assay). Photographs were taken at 100× magnification under a light microscope. Similar results were obtained in three independent experiments. (JPEG 64 kb)

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Lui, V.W.Y., Lau, C.P.Y., Ho, K. et al. Anti-invasion, anti-proliferation and anoikis-sensitization activities of lapatinib in nasopharyngeal carcinoma cells. Invest New Drugs 29, 1241–1252 (2011). https://doi.org/10.1007/s10637-010-9470-y

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