SPINK6 Promotes Metastasis of Nasopharyngeal Carcinoma via Binding and Activation of Epithelial Growth Factor Receptor

Cancer Res. 2017 Jan 15;77(2):579-589. doi: 10.1158/0008-5472.CAN-16-1281. Epub 2016 Sep 26.

Abstract

Nasopharyngeal carcinoma has the highest rate of metastasis among head and neck cancers, and distant metastasis is the major reason for treatment failure. The underlying molecular mechanisms of nasopharyngeal carcinoma metastasis are not fully understood. Here, we report the identification of serine protease inhibitor Kazal-type 6 (SPINK6) as a functional regulator of nasopharyngeal carcinoma metastasis via EGFR signaling. SPINK6 mRNA was upregulated in tumor and highly metastatic nasopharyngeal carcinoma cells. Immunohistochemical staining of 534 nasopharyngeal carcinomas revealed elevated SPINK6 expression as an independent unfavorable prognostic factor for overall, disease-free, and distant metastasis-free survival. Ectopic SPINK6 expression promoted in vitro migration and invasion as well as in vivo lymph node metastasis and liver metastasis of nasopharyngeal carcinoma cells, whereas silencing SPINK6 exhibited opposing effects. SPINK6 enhanced epithelial-mesenchymal transition by activating EGFR and the downstream AKT pathway. Inhibition of EGFR with a neutralizing antibody or erlotinib reversed SPINK6-induced nasopharyngeal carcinoma cell migration and invasion. Erlotinib also inhibited SPINK6-induced metastasis in vivo Notably, SPINK6 bound to the EGFR extracellular domain independent of serine protease-inhibitory activity. Overall, our results identified a novel EGFR-activating mechanism in which SPINK6 has a critical role in promoting nasopharyngeal carcinoma metastasis, with possible implications as a prognostic indicator in nasopharyngeal carcinoma patients. Cancer Res; 77(2); 579-89. ©2016 AACR.

MeSH terms

  • Carcinoma / metabolism
  • Carcinoma / mortality
  • Carcinoma / pathology*
  • Cell Movement / physiology
  • Disease-Free Survival
  • Epithelial-Mesenchymal Transition / physiology
  • ErbB Receptors / metabolism*
  • Female
  • Heterografts
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Immunoprecipitation
  • Kaplan-Meier Estimate
  • Lymphatic Metastasis / pathology
  • Male
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms / metabolism
  • Nasopharyngeal Neoplasms / mortality
  • Nasopharyngeal Neoplasms / pathology*
  • Neoplasm Invasiveness / pathology
  • Prognosis
  • Proportional Hazards Models
  • Proteinase Inhibitory Proteins, Secretory / metabolism*
  • Serine Peptidase Inhibitors, Kazal Type
  • Tissue Array Analysis

Substances

  • Proteinase Inhibitory Proteins, Secretory
  • SPINK6 protein, human
  • Serine Peptidase Inhibitors, Kazal Type
  • EGFR protein, human
  • ErbB Receptors