Merck
  • Home
  • Search Results
  • Activation of diverse signalling pathways by oncogenic PIK3CA mutations.

Activation of diverse signalling pathways by oncogenic PIK3CA mutations.

Nature communications (2014-09-24)
Xinyan Wu, Santosh Renuse, Nandini A Sahasrabuddhe, Muhammad Saddiq Zahari, Raghothama Chaerkady, Min-Sik Kim, Raja S Nirujogi, Morassa Mohseni, Praveen Kumar, Rajesh Raju, Jun Zhong, Jian Yang, Johnathan Neiswinger, Jun-Seop Jeong, Robert Newman, Maureen A Powers, Babu Lal Somani, Edward Gabrielson, Saraswati Sukumar, Vered Stearns, Jiang Qian, Heng Zhu, Bert Vogelstein, Ben Ho Park, Akhilesh Pandey
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Supelco
Hydrocortisone, VETRANAL®, analytical standard
Supelco
Hydrocortisone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Hydrocortisone, ≥98% (HPLC)
Sigma-Aldrich
Hydrocortisone, meets USP testing specifications
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-74, ascites fluid
Sigma-Aldrich
Hydrocortisone, BioReagent, suitable for cell culture
Sigma-Aldrich
Hydrocortisone, γ-irradiated, powder, BioXtra, suitable for cell culture
Hydrocortisone, British Pharmacopoeia (BP) Assay Standard
Hydrocortisone, European Pharmacopoeia (EP) Reference Standard
USP
Hydrocortisone, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
SAFC
L-Arginine
Supelco
L-Arginine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Arginine, 99%, FCC, FG
Sigma-Aldrich
Potassium phosphate monobasic, 99.99% trace metals basis
Sigma-Aldrich
Potassium phosphate monobasic, BioUltra, for molecular biology, anhydrous, ≥99.5% (T)
Sigma-Aldrich
Potassium phosphate monobasic, tested according to Ph. Eur., anhydrous
Sigma-Aldrich
L-Arginine, BioUltra, ≥99.5% (NT)
Sigma-Aldrich
L-Arginine, reagent grade, ≥98%
Sigma-Aldrich
L-Arginine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Potassium phosphate monobasic, for molecular biology, ≥98.0%
Sigma-Aldrich
Potassium phosphate monobasic, ReagentPlus®
Supelco
Monobasic Potassium Phosphate, Pharmaceutical Secondary Standard; Certified Reference Material
USP
Monobasic potassium phosphate, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Potassium phosphate monobasic, meets analytical specification of Ph. Eur., NF, E340, anhydrous, 98-100.5% (calc. to the dried substance)
Sigma-Aldrich
Potassium phosphate monobasic, ACS reagent, ≥99.0%
Hydrocortisone for peak identification, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Potassium phosphate monobasic, buffer substance, anhydrous, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., 99.5-100.5%
Sigma-Aldrich
Potassium phosphate monobasic, Vetec, reagent grade, 99%