Proteomic analysis reveals important role of 14-3-3σ in anoikis resistance of cholangiocarcinoma cells

Proteomics. 2013 Nov;13(21):3157-66. doi: 10.1002/pmic.201300219. Epub 2013 Oct 9.

Abstract

Cholangiocarcinoma (CCA), a high-prevalence cancer in Thailand, is highly metastatic and has high mortality rates. Anoikis resistance or the ability of cells to survive after detachment from extracellular matrix is a necessary property of metastatic cancer. Here, we report differential protein expression of an anoikis-resistant CCA cell line culture, under attachment conditions compared to nonattachment conditions, studied using 2DE coupled with protein identification by LC-MS/MS. Our data reveal the deregulation of proteins involved in stress response, cytoskeleton rearrangement, proapoptosis, cell proliferation, and glycolysis. Interestingly, 14-3-3σ (14-3-3 sigma) protein was intensely upregulated in detached CCA cells. Real-time RT-PCR analysis confirmed that only the sigma isotype was the most abundant transcript among 14-3-3 genes in CCA cells. Furthermore, silencing 14-3-3σ expression by small interfering RNA in CCA cells resulted in significantly increased percentage of cell death in detached culture. Our findings provide the first evidence showing that 14-3-3σ protein plays a crucial role in anoikis resistance of CCA cells. Therefore, 14-3-3σ might be a potential target in CCA therapy.

Keywords: 14-3-3 Sigma; Anoikis resistance; Cell biology; Cholangiocarcinoma; Metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 14-3-3 Proteins / genetics
  • 14-3-3 Proteins / metabolism*
  • Anoikis / genetics
  • Anoikis / physiology*
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / metabolism*
  • Cell Adhesion / genetics
  • Cell Adhesion / physiology
  • Cell Line, Tumor
  • Cell Shape
  • Cholangiocarcinoma / metabolism*
  • Electrophoresis, Gel, Two-Dimensional
  • Exoribonucleases / genetics
  • Exoribonucleases / metabolism*
  • Gene Knockdown Techniques
  • Humans
  • Proteome / analysis*
  • Proteome / chemistry
  • Proteome / metabolism
  • Proteomics / methods*
  • RNA, Small Interfering

Substances

  • 14-3-3 Proteins
  • Biomarkers, Tumor
  • Proteome
  • RNA, Small Interfering
  • Exoribonucleases
  • SFN protein, human