nonsense mediated decay resistant mutations are a source of expressed mutant proteins in colon cancer cell lines with microsatellite instability废话介导衰变耐药突变的突变蛋白在结肠癌细胞系表达微卫星不稳定.pdfVIP

Nonsense Mediated Decay Resistant Mutations Are a Source of Expressed Mutant Proteins in Colon Cancer Cell Lines with Microsatellite Instability 1,2,3 1 4 1 1 David S. Williams *, Matthew J. Bird , Robert N. Jorissen , Yen Lin Yu , Franscesa Walker , Hui Hua 1 1 1,4 Zhang , Edouard C. Nice , Antony W. Burgess * 1 Epithelial Biochemistry Laboratory, Ludwig Institute for Cancer Research, Melbourne Branch, Parkville, Victoria, Australia, 2 Department of Pathology, University of Melbourne, Parkville, Victoria, Australia, 3 Department of Anatomical Pathology, Melbourne Health, Parkville, Victoria, Australia, 4 Ludwig Colon Cancer Initiative Laboratory, Ludwig Institute for Cancer Research, Melbourne Branch, Parkville, Victoria, Australia Abstract Background: Frameshift mutations in microsatellite instability high (MSI-High) colorectal cancers are a potential source of targetable neo-antigens. Many nonsense transcripts are subject to rapid degradation due to nonsense-mediated decay (NMD), but nonsense transcripts with a cMS in the last exon or near the last exon-exon junction have intrinsic resistance to nonsense-mediated decay (NMD). NMD-resistant transcripts are therefore a likely source of expressed mutant proteins in MSI-High tumours. Methods: Using antibodies to the conserved N-termini of predicted mutant proteins, we analysed MSI-High colorectal cancer cell lines for examples of naturally expressed mutant proteins arising from frameshift mutations in coding microsatellites (cMS) by immunoprecipitation and Western Blot experiments. Detected mutant protein bands from NMD-