title: Genome-Wide Overexpression Screen Identifies Genes Able to Bypass p16-Mediated Senescence in Melanoma.
abstract: Malignant melanomas often arise from nevi, which result from initial oncogene-induced hyperproliferation of melanocytes that are maintained in a CDKN2A/p16-mediated senescent state. Thus, genes that can bypass this senescence barrier are likely to contribute to melanoma development. We have performed a gain-of-function screen of 17,030 lentivirally expressed human open reading frames (ORFs) in a melanoma cell line containing an inducible p16 construct to identify such genes. Genes known to bypass p16-induced senescence arrest, including the human papilloma virus 18 E7 gene ( HPV18E7), and genes such as the p16-binding CDK6 with expected functions, as well as panel of novel genes, were identified, including high-mobility group box (HMGB) proteins. A number of these were further validated in two other models of p16-induced senescence. Tissue immunohistochemistry demonstrated higher levels of CDK6 in primary melanomas compared with normal skin and nevi. Reduction of CDK6 levels drove melanoma cells expressing functional p16 into senescence, demonstrating its contribution to bypass senescence.
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Won Jae
authors.initials: WJ
authors.lastname: Lee
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.|Mater Research Institute, The University of Queensland, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Dubravka
authors.initials: D
authors.lastname: Škalamera
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.|Mater Research Institute, The University of Queensland, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Mareike
authors.initials: M
authors.lastname: Dahmer-Heath
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.|Queensland Brain Institute, The University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Konstanin
authors.initials: K
authors.lastname: Shakhbazov
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Max V
authors.initials: MV
authors.lastname: Ranall
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Carly
authors.initials: C
authors.lastname: Fox
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Duncan
authors.initials: D
authors.lastname: Lambie
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.|Mater Research Institute, The University of Queensland, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Stevenson
authors.affiliation: 2 Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Yaswen
authors.affiliation: 3 School of Pharmacy, The University of Queensland, Brisbane, Queensland, Australia.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Gonda
authors.affiliation: 1 The University of Queensland Diamantina Institute, Translational Research Institute, Brisbane, Queensland, Australia.|Mater Research Institute, The University of Queensland, Translational Research Institute, Brisbane, Queensland, Australia.
authors.forename: Brian
authors.initials: B
authors.lastname: Gabrielli
mesh_terms: D059447:Cell Cycle Checkpoints; D045744:Cell Line, Tumor; D016922:Cellular Senescence; D051361:Cyclin-Dependent Kinase 6 / Q000235:genetics* / Q000378:metabolism; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D015894:Genome, Human; D057809:HEK293 Cells; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008544:Melanocytes / Q000378:metabolism* / Q000473:pathology; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009506:Nevus / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames; D012878:Skin Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D004268:DNA-Binding Proteins; C052633:E7 protein, Human papillomavirus type 18; D024002:HMGB Proteins; D009856:Oncogene Proteins, Viral; C495911:CDK6 protein, human; D051361:Cyclin-Dependent Kinase 6
keywords: CDK6; high-content imaging; overexpression screening; p16; senescence