title: Uptake by human glioma cell lines and biological effects of a peptide-nucleic acids targeting miR-221.
abstract: MicroRNAs are a family of small noncoding RNAs regulating gene expression by sequence-selective mRNA targeting, leading to a translational repression or mRNA degradation. The oncomiR miR-221 is highly expressed in human gliomas, as confirmed in this study in samples of low and high grade gliomas, as well in the cell lines U251, U373 and T98G. In order to alter the biological functions of miR-221, a peptide nucleic acid targeting miR-221 (R8-PNA-a221) was produced, bearing a oligoarginine peptide (R8) to facilitate uptake by glioma cells. The effects of R8-PNA-a221 were analyzed in U251, U373 and T98G glioma cells and found to strongly inhibit miR-221. In addition, the effects of R8-PNA-a221 on p27(Kip1) (a target of miR-221) were analyzed in U251 and T98G cells by RT-qPCR and by Western blotting. No change of p27(Kip1) mRNA content occurs in U251 cells in the presence of PNA-a221 (lacking the R8 peptide), whereas significant increase of p27(Kip1) mRNA was observed with the R8-PNA-a221. These data were confirmed by Western blot assay. A clear increment of p27(Kip1) protein expression in the samples treated with R8-PNA-a221 was detected. In addition, R8-PNA-a221 was found able to increase TIMP3 expression (another target of miR-221) in T98G cells. These results suggest that PNAs against oncomiRNA miR-221 might be proposed for experimental treatment of human gliomas.
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Brognara
authors.forename: Enrica
authors.initials: E
authors.lastname: Fabbri
authors.forename: Elena
authors.initials: E
authors.lastname: Bazzoli
authors.forename: Giulia
authors.initials: G
authors.lastname: Montagner
authors.forename: Claudio
authors.initials: C
authors.lastname: Ghimenton
authors.forename: Albino
authors.initials: A
authors.lastname: Eccher
authors.forename: Cinzia
authors.initials: C
authors.lastname: Cantù
authors.forename: Alex
authors.initials: A
authors.lastname: Manicardi
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Bianchi
authors.forename: Alessia
authors.initials: A
authors.lastname: Finotti
authors.forename: Giulia
authors.initials: G
authors.lastname: Breveglieri
authors.forename: Monica
authors.initials: M
authors.lastname: Borgatti
authors.forename: Roberto
authors.initials: R
authors.lastname: Corradini
authors.forename: Valentino
authors.initials: V
authors.lastname: Bezzerri
authors.forename: Giulio
authors.initials: G
authors.lastname: Cabrini
authors.forename: Roberto
authors.initials: R
authors.lastname: Gambari
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D005910:Glioma / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D020135:Peptide Nucleic Acids / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D017304:Annexin A5; D050760:Cyclin-Dependent Kinase Inhibitor p27; D035683:MicroRNAs; D020135:Peptide Nucleic Acids; C525709:MIR221, human