title: Long non-coding RNA containing ultraconserved genomic region 8 promotes bladder cancer tumorigenesis.
abstract: Ultraconserved regions (UCRs) have been shown to originate non-coding RNA transcripts (T-UCRs) that have different expression profiles and play functional roles in the pathophysiology of multiple cancers. The relevance of these functions to the pathogenesis of bladder cancer (BlCa) is speculative. To elucidate this relevance, we first used genome-wide profiling to evaluate the expression of T-UCRs in BlCa tissues. Analysis of two datasets comprising normal bladder tissues and BlCa specimens with a custom T-UCR microarray identified ultraconserved RNA (uc.) 8+ as the most upregulated T-UCR in BlCa tissues, although its expression was lower than in pericancerous bladder tissues. These results were confirmed on BlCa tissues by real-time PCR and by in situ hybridization. Although uc.8+ is located within intron 1 of CASZ1, a zinc-finger transcription factor, the transcribed non-coding RNA encoding uc.8+ is expressed independently of CASZ1. In vitro experiments evaluating the effects of uc.8+ silencing, showed significantly decreased capacities for cancer cell invasion, migration, and proliferation. From this, we proposed and validated a model of interaction in which uc.8+ shuttles from the nucleus to the cytoplasm of BlCa cells, interacts with microRNA (miR)-596, and cooperates in the promotion and development of BlCa. Using computational analysis, we investigated the miR-binding domain accessibility, as determined by base-pairing interactions within the uc.8+ predicted secondary structure, RNA binding affinity, and RNA species abundance in bladder tissues and showed that uc.8+ is a natural decoy for miR-596. Thus uc.8+ upregulation results in increased expression of MMP9, increasing the invasive potential of BlCa cells. These interactions between evolutionarily conserved regions of DNA suggest that natural selection has preserved this potentially regulatory layer that uses RNA to modulate miR levels, opening up the possibility for development of useful markers for early diagnosis and prognosis as well as for development of new RNA-based cancer therapies.
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Olivieri
authors.affiliation: Division of Urology, European Institute of Oncology, Milan, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Ferro
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Terreri
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.|Bio-Ker S.r.l., MultiMedica S.p.A., Naples, Italy.
authors.forename: Montano
authors.initials: M
authors.lastname: Durso
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", Naples, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Romanelli
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", Naples, Italy.
authors.forename: Concetta
authors.initials: C
authors.lastname: Avitabile
authors.affiliation: Division of Urology, European Institute of Oncology, Milan, Italy.
authors.forename: Ottavio
authors.initials: O
authors.lastname: De Cobelli
authors.affiliation: Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, Second University of Naples, Naples, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Messere
authors.affiliation: Department of Public Health, University of Naples "Federico II", Naples, Italy.
authors.forename: Dario
authors.initials: D
authors.lastname: Bruzzese
authors.affiliation: Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) S.r.l. IRCCS, Gene Therapy Unit, Meldola, Italy.
authors.forename: Ivan
authors.initials: I
authors.lastname: Vannini
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", Naples, Italy.
authors.forename: Luciana
authors.initials: L
authors.lastname: Marinelli
authors.affiliation: Department of Pharmacy, University of Naples "Federico II", Naples, Italy.
authors.forename: Ettore
authors.initials: E
authors.lastname: Novellino
authors.affiliation: Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Fondazione IRCCS SDN, Naples, Italy.
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: Incoronato
authors.affiliation: Department of Advanced Biomedical Sciences, University of Naples "Federico II", Naples, Italy.
authors.forename: Gennaro
authors.initials: G
authors.lastname: Ilardi
authors.affiliation: Department of Advanced Biomedical Sciences, University of Naples "Federico II", Naples, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Staibano
authors.affiliation: Division of Urology, IRCS National Tumor Institute, Naples, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Marra
authors.affiliation: Department of Physical and Mental Health and Preventive Medicine, Section of Pathology, Second University of Naples, Naples, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Franco
authors.affiliation: Division of Urology, IRCS National Tumor Institute, Naples, Italy.
authors.forename: Sisto
authors.initials: S
authors.lastname: Perdonà
authors.affiliation: Department of Translational Medical Sciences, University of Naples "Federico II", Naples, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Terracciano
authors.affiliation: Department of Pathology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Bogdan
authors.initials: B
authors.lastname: Czerniak
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.
authors.forename: Giovanna L
authors.initials: GL
authors.lastname: Liguori
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.
authors.forename: Vincenza
authors.initials: V
authors.lastname: Colonna
authors.affiliation: Department of Pediatrics and Molecular Microbiology & Immunology, Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, Children's Center for Cancer and Blood Diseases and The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA, USA.
authors.forename: Muller
authors.initials: M
authors.lastname: Fabbri
authors.affiliation: Institute of Protein Biochemistry, National Research Council (CNR), Naples, Italy.
authors.forename: Ferdinando
authors.initials: F
authors.lastname: Febbraio
authors.affiliation: Department of Experimental Therapeutics and The Center for RNA Interference and Non-Coding RNA, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: George A
authors.initials: GA
authors.lastname: Calin
authors.affiliation: Institute of Genetics and Biophysics "A. Buzzati Traverso", National Research Council (CNR), Naples, Italy.
authors.forename: Amelia
authors.initials: A
authors.lastname: Cimmino
mesh_terms: D000368:Aged; D017209:Apoptosis; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics; D002465:Cell Movement; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000235:genetics* / Q000473:pathology; D017124:Conserved Sequence / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D022661:RNA, Untranslated / Q000235:genetics*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D022661:RNA, Untranslated
keywords: CASZ1; MMP9; T-UCR; bladder cancer; microRNA