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Investigation into the influence of the P13Ka (H1047R) mutation on the bioenergetic dependency of a cell
Rebecca Foster, Sue Rigby, Ruth Feltell, Suzanne Mead, Cindy Christopherson, John Sninsky, Shirley Kwok and Chris Torrance
Horizon Discovery Services Ltd, Cambridge, UK, Celera Corp, Alameda, CA, Horizon Discovery Ltd, Cambridge, UK
Sponsored by Horizon Discovery Ltd.
Published in July 2011 2011 (p.7) Sponsored,vendor-submitted protocol
During tumorigenesis, cells acquire mutations that confer selective advantages for growth. These mutations may reprogram cellular metabolism to meet the requirements of rapid proliferation and survival in an environment of fluctuating and scarce nutrients. Phosphoinositide-3-kinases (P13Ks) play key roles in fundamental cellular processes and are frequently mutated in a broad range of cancers. The aim of this study was to investigate how a single endogenous activating mutation in P13Ka, H1047R, affects the metabolic programming of human breast epithelial cells.
We utilized an X-MAN (X-Mutant And Normal) isogenic cell line derived from MCF10A mammary epithelial cells, which harbours a single P13Ka mutation introduced via rAAV-mediated homologous recombination, and compared it to a matched parental cell line with a wild type allele. This system allows use of endogenous promoters thereby more accurately recapitulating the genomic architecture in human tumours.
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