Comparative proteomic analysis of oral squamous cell carcinoma and adjacent non-tumour tissue from Thailand

Arch Oral Biol. 2013 Nov;58(11):1677-85. doi: 10.1016/j.archoralbio.2013.08.002. Epub 2013 Aug 13.

Abstract

Objective: The study was aimed at analysing and identifying the proteins that are differentially expressed in oral squamous cell carcinoma (OSCC) compared to adjacent non-tumour tissue.

Materials and methods: Two-dimensional (2D) sodium dodecyl sulphate-polyacrylamide gel electrophoresis accompanied by mass spectrometry (matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometry and liquid chromatography-tandem mass spectrometry) was used to analyse and identify the differentially expressed proteins in 10 pairs of tumours and adjacent non-tumour tissues from five cases of early-stage and five cases of late-stage OSCC. The statistical differences of the protein spots were analysed by the Wilcoxon signed-rank test. A validation study using immunohistochemistry and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was performed.

Results: A total of 68 proteins (63 up-regulated, five down-regulated) were differentially expressed in early-stage disease, and 39 proteins (37 up-regulated, two down-regulated) were significantly altered in late-stage disease. Among these, 14 proteins were altered in both groups. A total of 44 proteins were identified, including heat shock proteins (HSPs: Hsp90, HSPA5 and HSPA8), keratins (K1, K6A and K17), tubulin, cofilin 1, 14-3-3σ and metabolic enzymes. These proteins are involved in various cellular processes essential for cell growth, survival and cell migration. The validation study on α-tubulin and 14-3-3σ using immunohistochemistry and KIAA1199 expression using real-time RT-PCR confirmed the results in proteomics analysis.

Conclusions: The study identified many proteins, both known and unknown, for cancer cell processes. At least two proteins, KIAA1199 and Horf6, are novel for oral cancer.

Keywords: Immunohistochemistry; Mass spectrometry; Oral squamous cell carcinoma; Proteomics; Quantitative real-time RT-PCR; Two-dimensional gel electrophoresis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Biomarkers, Tumor / analysis
  • Biomarkers, Tumor / genetics*
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / metabolism
  • Down-Regulation*
  • Electrophoresis, Gel, Two-Dimensional
  • Endoplasmic Reticulum Chaperone BiP
  • Gene Expression Regulation, Neoplastic / physiology*
  • Heat-Shock Proteins / analysis
  • Humans
  • Hyaluronoglucosaminidase
  • Immunohistochemistry
  • Keratins / analysis
  • Mouth Neoplasms / genetics*
  • Mouth Neoplasms / metabolism
  • Peroxiredoxin VI / analysis
  • Proteins / analysis
  • Proteomics / methods*
  • Real-Time Polymerase Chain Reaction
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Thailand
  • Tubulin / analysis
  • Up-Regulation*

Substances

  • Biomarkers, Tumor
  • Endoplasmic Reticulum Chaperone BiP
  • HSPA5 protein, human
  • Heat-Shock Proteins
  • Proteins
  • Tubulin
  • Keratins
  • Peroxiredoxin VI
  • CEMIP protein, human
  • Hyaluronoglucosaminidase