title: Nucleic Acid Targeted Therapy: G4 Oligonucleotides Downregulate HRAS in Bladder Cancer Cells through a Decoy Mechanism.
abstract: In a previous study we have demonstrated that two neighboring G-quadruplexes, hras-1 and hras-2, located immediately upstream of the major transcription start site of HRAS, bind MAZ, a nuclear factor that activates transcription (Cogoi, S.; et al. Nucl. Acid Res. 2014, 42, 8379). For the present study we have designed G4 oligonucleotides with anthraquinone insertions and locked nucleic acids (LNA) modifications mimicking quadruplex hras-1. Luciferase, qRT-PCR, and Western blot data demonstrate that these constructs efficiently down regulate HRAS in T24 bladder cancer cells. The inhibitory efficiency of the G4 oligonucleotides correlates with their nuclease resistance in the cell environment. By chromatin immunoprecipitation we show that the association of MAZ to the HRAS promoter is strongly attenuated by the designed G4 oligonucleotides, thus suggesting that these constructs behave with a decoy mechanism.
authors.affiliation: Department of Medical and Biological Sciences, University of Udine , 33100 Udine, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Miglietta
authors.affiliation: Nucleic Acid Center, Institute of Physics and Chemistry, University of Southern Denmark , DK-5230 Odense M, Denmark.
authors.forename: Alaa S
authors.initials: AS
authors.lastname: Gouda
authors.affiliation: Department of Medical and Biological Sciences, University of Udine , 33100 Udine, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Cogoi
authors.affiliation: Nucleic Acid Center, Institute of Physics and Chemistry, University of Southern Denmark , DK-5230 Odense M, Denmark.
authors.forename: Erik B
authors.initials: EB
authors.lastname: Pedersen
authors.affiliation: Department of Medical and Biological Sciences, University of Udine , 33100 Udine, Italy.
authors.forename: Luigi E
authors.initials: LE
authors.lastname: Xodo
keywords: G4-oligonucleotides; HRAS; T24 bladder cancer cells; anthraquinone insertions; decoy mechanism