The E545K mutation of PIK3CA promotes gallbladder carcinoma progression through enhanced binding to EGFR

J Exp Clin Cancer Res. 2016 Jun 18;35(1):97. doi: 10.1186/s13046-016-0370-7.

Abstract

Background: Gallbladder carcinoma (GBC) is the most common malignancy of the bile duct and patients with GBC have extremely poor prognoses. PIK3CA, which encodes the phosphoinositide 3-kinase (PI3K) subunit p110α, is frequently mutated in many cancers, including GBC. The function of the E545K mutation in GBC is not fully understood.

Methods: E545K mutation was determined in human GBC tissues by targeted sequencing. The effects of E545K mutation and PI3K selective inhibitor, A66 on GBC cells were evaluated using Cell Counting Kit-8 (CCK-8) cell Viability and transwell assays. The mechanisms of E545K mutation and A66 were analyzed by western blot and co-immunoprecipitation (Co-IP) assay. Subcutaneous xenograft models in nude mice were employed to evaluate the role of E545K mutation and A66 in GBC progression.

Results: The rate of PIK3CA E545K mutation in GBC patients was 6.15 %. And the survival of GBC patients was correlated with E545K mutation significantly (P < 0.05). The E545K mutation promoted proliferation, migration and invasion of GBC cells in vitro and tumor proliferation in vivo. A66 suppressed proliferation of GBC cells in vitro and tumor proliferation in vivo.

Conclusion: The prognoses of patients with E545K mutation were worse than patients without this mutation. The E545K mutation promoted GBC progression through enhanced binding to EGFR and activating downstream akt activity. The PI3K selective inhibitor, A66, suppressed gallbladder carcinoma proliferation.

Keywords: Cancer; EGFR; Gallbladder carcinoma; PI3K.

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell Movement
  • Cell Proliferation
  • Class I Phosphatidylinositol 3-Kinases / antagonists & inhibitors
  • Class I Phosphatidylinositol 3-Kinases / genetics*
  • Class I Phosphatidylinositol 3-Kinases / metabolism*
  • Disease Progression
  • ErbB Receptors / metabolism*
  • Female
  • Gallbladder Neoplasms / genetics
  • Gallbladder Neoplasms / pathology*
  • Humans
  • Male
  • Mice
  • Mice, Nude
  • Mutation*
  • Neoplasm Invasiveness
  • Neoplasm Transplantation
  • Prognosis
  • Proline / analogs & derivatives
  • Proline / pharmacology
  • Protein Binding
  • Signal Transduction
  • Survival Analysis
  • Thiazoles / pharmacology

Substances

  • N1-(2-(t-butyl)-4'-methyl-(4,5'-bithiazol)- 2'-yl)pyrrolidine-1,2-dicarboxamide
  • Thiazoles
  • Proline
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • EGFR protein, human
  • ErbB Receptors