title: Differential reprogramming of isogenic colorectal cancer cells by distinct activating KRAS mutations.
abstract: Oncogenic mutations of Ras at codons 12, 13, or 61, that render the protein constitutively active, are found in ∼ 16% of all cancer cases. Among the three major Ras isoforms, KRAS is the most frequently mutated isoform in cancer. Each Ras isoform and tumor type displays a distinct pattern of codon-specific mutations. In colon cancer, KRAS is typically mutated at codon 12, but a significant fraction of patients have mutations at codon 13. Clinical data suggest different outcomes and responsiveness to treatment between these two groups. To investigate the differential effects upon cell status associated with KRAS mutations we performed a quantitative analysis of the proteome and phosphoproteome of isogenic SW48 colon cancer cell lines in which one allele of the endogenous gene has been edited to harbor specific KRAS mutations (G12V, G12D, or G13D). Each mutation generates a distinct signature, with the most variability seen between G13D and the codon 12 KRAS mutants. One notable example of specific up-regulation in KRAS codon 12 mutant SW48 cells is provided by the short form of the colon cancer stem cell marker doublecortin-like Kinase 1 (DCLK1) that can be reversed by suppression of KRAS.
authors.affiliation: Division of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool , Crown Street, Liverpool L69 3BX, United Kingdom.
authors.forename: Dean E
authors.initials: DE
authors.lastname: Hammond
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Mageean
authors.forename: Emma V
authors.initials: EV
authors.lastname: Rusilowicz
authors.forename: Julie A
authors.initials: JA
authors.lastname: Wickenden
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Clague
authors.forename: Ian A
authors.initials: IA
authors.lastname: Prior
mesh_terms: D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics / Q000473:pathology*; D000089742:Doublecortin-Like Kinases; D011905:Genes, ras; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D009154:Mutation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D040901:Proteomics; D015854:Up-Regulation
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C117347:DCLK1 protein, human; D000089742:Doublecortin-Like Kinases; D017346:Protein Serine-Threonine Kinases
keywords: Ras; SILAC phosphoproteomics; colorectal cancer; isogenic cells; signaling