Targeting polo-like kinase 1 enhances radiation efficacy for head-and-neck squamous cell carcinoma

Int J Radiat Oncol Biol Phys. 2010 May 1;77(1):253-60. doi: 10.1016/j.ijrobp.2009.11.027.

Abstract

Purpose: To investigate the efficacy of targeting polo-like kinase 1 (Plk1) combined with ionizing radiotherapy (RT) for head-and-neck squamous cell carcinoma (HNSCC).

Methods and materials: Polo-like kinase 1 messenger ribonucleic acid (mRNA) was targeted by small interfering RNA (siRNA) transfection into the FaDu HNSCC cell line; reduction was confirmed using quantitative real-time polymerase chain reaction. The cellular effects were assessed using [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium], clonogenic, flow cytometric, and caspase assays. In vivo efficacy of siPlk1 was evaluated using mouse xenograft models.

Results: Small interfering Plk1 significantly decreased Plk1 mRNA expression, while also increasing cyclin B1 and p21(Waf1/CIP1) mRNA levels after 24 h. This depletion resulted in a time-dependent increase in FaDu cytotoxicity, which was enhanced by the addition of RT. Flow cytometric and caspase assays demonstrated progressive apoptosis, DNA double-strand breaks (gamma-H2AX), G2/M arrest, and activation of caspases 3 and 7. Implantation of siPlk1-treated FaDu cells in severe combined immunodeficient mice delayed tumor formation, and systemic administration of siPlk1 inhibited tumor growth enhanced by RT.

Conclusions: These data demonstrate the suitability of Plk1 as a potential therapeutic target for HNSCC, because Plk1 depletion resulted in significant cytotoxicity in vitro and abrogated tumor-forming potential in vivo. The effects of Plk1 depletion were enhanced with the addition of RT, indicating that Plk1 represents an important potential radiation sensitizer for HNSCC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Carcinoma, Squamous Cell / drug therapy
  • Carcinoma, Squamous Cell / radiotherapy*
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Cycle Proteins / antagonists & inhibitors*
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Cyclin B1 / metabolism
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Female
  • Head and Neck Neoplasms / drug therapy
  • Head and Neck Neoplasms / radiotherapy*
  • Histones / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Neoplasm Proteins / antagonists & inhibitors*
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Polo-Like Kinase 1
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Proto-Oncogene Proteins / antagonists & inhibitors*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • RNA, Messenger / antagonists & inhibitors*
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / therapeutic use*
  • Tetrazolium Salts
  • Thiazoles
  • Transfection
  • Tumor Stem Cell Assay / methods

Substances

  • CDKN1A protein, human
  • Cell Cycle Proteins
  • Cyclin B1
  • Cyclin-Dependent Kinase Inhibitor p21
  • H2AX protein, human
  • Histones
  • Neoplasm Proteins
  • Proto-Oncogene Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • Tetrazolium Salts
  • Thiazoles
  • 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium
  • Protein Serine-Threonine Kinases
  • Caspase 3
  • Caspase 7