title: The malignant phenotype in breast cancer is driven by eIF4A1-mediated changes in the translational landscape.
abstract: Human mRNA DeXD/H-box helicases are ubiquitous molecular motors that are required for the majority of cellular processes that involve RNA metabolism. One of the most abundant is eIF4A, which is required during the initiation phase of protein synthesis to unwind regions of highly structured mRNA that would otherwise impede the scanning ribosome. Dysregulation of protein synthesis is associated with tumorigenesis, but little is known about the detailed relationships between RNA helicase function and the malignant phenotype in solid malignancies. Therefore, immunohistochemical analysis was performed on over 3000 breast tumors to investigate the relationship among expression of eIF4A1, the helicase-modulating proteins eIF4B, eIF4E and PDCD4, and clinical outcome. We found eIF4A1, eIF4B and eIF4E to be independent predictors of poor outcome in ER-negative disease, while in contrast, the eIF4A1 inhibitor PDCD4 was related to improved outcome in ER-positive breast cancer. Consistent with these data, modulation of eIF4A1, eIF4B and PCDC4 expression in cultured MCF7 cells all restricted breast cancer cell growth and cycling. The eIF4A1-dependent translatome of MCF7 cells was defined by polysome profiling, and was shown to be highly enriched for several classes of oncogenic genes, including G-protein constituents, cyclins and protein kinases, and for mRNAs with G/C-rich 5'UTRs with potential to form G-quadruplexes and with 3'UTRs containing microRNA target sites. Overall, our data show that dysregulation of mRNA unwinding contributes to the malignant phenotype in breast cancer via preferential translation of a class of genes involved in pro-oncogenic signaling at numerous levels. Furthermore, immunohistochemical tests are promising biomarkers for tumors sensitive to anti-helicase therapies.
authors.affiliation: Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: A
authors.initials: A
authors.lastname: Modelska
authors.affiliation: 1] Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK [2] Department of Haematology, University of Cambridge, NHS Blood and Transplant, Long Road, Cambridge CB2 0PT, UK.
authors.forename: E
authors.initials: E
authors.lastname: Turro
authors.affiliation: Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: R
authors.initials: R
authors.lastname: Russell
authors.affiliation: Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: J
authors.initials: J
authors.lastname: Beaton
authors.affiliation: Medical Research Council Toxicology Unit, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: T
authors.initials: T
authors.lastname: Sbarrato
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham NG7 2RD, UK.
authors.forename: K
authors.initials: K
authors.lastname: Spriggs
authors.affiliation: Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: J
authors.initials: J
authors.lastname: Miller
authors.affiliation: 1] Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK [2] Department of Haematology, University of Cambridge, NHS Blood and Transplant, Long Road, Cambridge CB2 0PT, UK [3] Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge CB2 0QQ, UK.
authors.forename: S
authors.initials: S
authors.lastname: Gräf
authors.affiliation: 1] Cambridge Experimental Cancer Medicine Centre (ECMR) and NIHR Cambridge Biomedical Research Centre, Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 0QQ, UK [2] Cambridge Breast Unit and Cambridge University Hospitals NHS Foundation Trust, Hills Road, Cambridge CB2 0QQ, UK.
authors.forename: E
authors.initials: E
authors.lastname: Provenzano
authors.affiliation: Strangeways Research Institute, 2 Worts' Causeway, Cambridge CB1 8RN, UK.
authors.forename: F
authors.initials: F
authors.lastname: Blows
authors.affiliation: 1] Cambridge Experimental Cancer Medicine Centre (ECMR) and NIHR Cambridge Biomedical Research Centre, Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 0QQ, UK [2] Strangeways Research Institute, 2 Worts' Causeway, Cambridge CB1 8RN, UK.
authors.forename: P
authors.initials: P
authors.lastname: Pharoah
authors.affiliation: 1] Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK [2] Cambridge Experimental Cancer Medicine Centre (ECMR) and NIHR Cambridge Biomedical Research Centre, Cambridge University Hospitals NHS Foundation Trust, Cambridge CB2 0QQ, UK.
authors.forename: C
authors.initials: C
authors.lastname: Caldas
authors.affiliation: 1] Department of Oncology and Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK [2] Medical Research Council Toxicology Unit, Lancaster Road, Leicester LE1 9HN, UK.
authors.forename: J
authors.initials: J
authors.lastname: Le Quesne
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D017209:Apoptosis / Q000235:genetics; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004265:DNA Helicases / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics; D004721:Endoplasmic Reticulum / Q000378:metabolism; D039601:Eukaryotic Initiation Factor-4A / Q000378:metabolism*; D039642:Eukaryotic Initiation Factors / Q000378:metabolism; D005260:Female; D052138:Genes, Neoplasm; D006801:Humans; D053208:Kaplan-Meier Estimate; D035683:MicroRNAs / Q000378:metabolism; D008875:Middle Aged; D008969:Molecular Sequence Data; D015999:Multivariate Analysis; D010641:Phenotype; D016016:Proportional Hazards Models; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D016896:Treatment Outcome
chemical_list: D020121:5' Untranslated Regions; D051017:Apoptosis Regulatory Proteins; D039642:Eukaryotic Initiation Factors; D035683:MicroRNAs; C114487:PDCD4 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011960:Receptors, Estrogen; C031799:eIF-4B; D039601:Eukaryotic Initiation Factor-4A; D004265:DNA Helicases