Activation of diverse signalling pathways by oncogenic PIK3CA mutations

Nat Commun. 2014 Sep 23:5:4961. doi: 10.1038/ncomms5961.


The PIK3CA gene is frequently mutated in human cancers. Here we carry out a SILAC-based quantitative phosphoproteomic analysis using isogenic knockin cell lines containing 'driver' oncogenic mutations of PIK3CA to dissect the signalling mechanisms responsible for oncogenic phenotypes induced by mutant PIK3CA. From 8,075 unique phosphopeptides identified, we observe that aberrant activation of PI3K pathway leads to increased phosphorylation of a surprisingly wide variety of kinases and downstream signalling networks. Here, by integrating phosphoproteomic data with human protein microarray-based AKT1 kinase assays, we discover and validate six novel AKT1 substrates, including cortactin. Through mutagenesis studies, we demonstrate that phosphorylation of cortactin by AKT1 is important for mutant PI3K-enhanced cell migration and invasion. Our study describes a quantitative and global approach for identifying mutation-specific signalling events and for discovering novel signalling molecules as readouts of pathway activation or potential therapeutic targets.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chromatography, Liquid
  • Class I Phosphatidylinositol 3-Kinases
  • Cortactin / metabolism*
  • DNA Primers / genetics
  • Fluorescent Antibody Technique
  • Gene Knock-In Techniques
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Mutagenesis, Site-Directed
  • Mutation / genetics
  • Phosphatidylinositol 3-Kinases / genetics*
  • Phosphoproteins / genetics*
  • Proteomics
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA, Small Interfering / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics
  • Signal Transduction / physiology*
  • Tandem Mass Spectrometry


  • Cortactin
  • DNA Primers
  • Phosphoproteins
  • RNA, Small Interfering
  • Phosphatidylinositol 3-Kinases
  • Class I Phosphatidylinositol 3-Kinases
  • PIK3CA protein, human
  • AKT1 protein, human
  • Proto-Oncogene Proteins c-akt