title: Loss-of-function screening to identify miRNAs involved in senescence: tumor suppressor activity of miRNA-335 and its new target CARF.
abstract: Significance of microRNAs (miRs), small non-coding molecules, has been implicated in a variety of biological processes. Here, we recruited retroviral insertional mutagenesis to obtain induction of an arbitrary noncoding RNAs, and coupled it with a cell based loss-of-function (5-Aza-2'-deoxycytidine (5Aza-dC)-induced senescence bypass) screening system. Cells that escaped 5-Aza-dC-induced senescence were subjected to miR-microarray analysis with respect to the untreated control. We identified miR-335 as one of the upregulated miRs. In order to characterize the functional significance, we overexpressed miR-335 in human cancer cells and found that it caused growth suppression. We demonstrate that the latter accounted for inhibition of 5-Aza-dC incorporation into the cell genome, enabling them to escape from induction of senescence. We also report that CARF (Collaborator of ARF) is a new target of miR-335 that regulates its growth suppressor function by complex crosstalk with other proteins including p16(INK4A), pRB, HDM2 and p21(WAF1).
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.|Graduate School of Life &Environmental Sciences, University of Tsukuba, Japan.
authors.forename: Yue
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.|Institute of Laboratory Animal Science, Chinese Academy of Medical Science (CAMS) &Comparative Medicine Center, Peking Union Medical College (PUMC), China.
authors.forename: Ran
authors.initials: R
authors.lastname: Gao
authors.affiliation: Department of Molecular Virology, Immunology &Medical Genetics, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Zeenia
authors.initials: Z
authors.lastname: Kaul
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.|Graduate School of Life &Environmental Sciences, University of Tsukuba, Japan.
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Kato
authors.affiliation: Graduate School of Life &Environmental Sciences, University of Tsukuba, Japan.
authors.forename: Zhenya
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Molecular Virology, Immunology &Medical Genetics, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Joanna
authors.initials: J
authors.lastname: Groden
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.
authors.forename: Sunil C
authors.initials: SC
authors.lastname: Kaul
authors.affiliation: Drug Discovery and Assets Innovation Lab, DBT-AIST International Laboratory for Advanced Biomedicine (DAILAB), Biomedical Research Institute, National Institute of Advanced Industrial Science &Technology (AIST), Tsukuba-305 8565, Japan.
authors.forename: Renu
authors.initials: R
authors.lastname: Wadhwa
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D016922:Cellular Senescence / Q000187:drug effects; D000077209:Decitabine; D016147:Genes, Tumor Suppressor; D019942:Genes, p16; D064593:Heterografts; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C539004:CDKN2AIP protein, human; C556347:MIRN335 microRNA, human; D035683:MicroRNAs; D016601:RNA-Binding Proteins; D016160:Retinoblastoma Protein; D000077209:Decitabine; D001374:Azacitidine