title: Effect of Polypurine Reverse Hoogsteen Hairpins on Relevant Cancer Target Genes in Different Human Cell Lines.
abstract: We studied the ability of polypurine reverse Hoogsteen hairpins (PPRHs) to silence a variety of relevant cancer-related genes in several human cell lines. PPRHs are hairpins formed by two antiparallel polypurine strands bound by intramolecular Hoogsteen bonds linked by a pentathymidine loop. These hairpins are able to bind to their target DNA sequence through Watson-Crick bonds producing specific silencing of gene expression. We designed PPRHs against the following genes: BCL2, TOP1, mTOR, MDM2, and MYC and tested them for mRNA levels, cytotoxicity, and apoptosis in prostate, pancreas, colon, and breast cancer cell lines. Even though all PPRHs were effective, the most remarkable results were obtained with those against BCL2 and mammalian target of rapamycin (mTOR) in decreasing cell survival and mRNA levels and increasing apoptosis in prostate, colon, and pancreatic cancer cells. In the case of TOP1, MDM2, and MYC, their corresponding PPRHs produced a strong effect in decreasing cell viability and mRNA levels and increasing apoptosis in breast cancer cells. Thus, we confirm that the PPRH technology is broadly useful to silence the expression of cancer-related genes as demonstrated using target genes involved in metabolism (DHFR), proliferation (mTOR), DNA topology (TOP1), lifespan and senescence (telomerase), apoptosis (survivin, BCL2), transcription factors (MYC), and proto-oncogenes (MDM2).
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Xenia
authors.initials: X
authors.lastname: Villalobos
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Laura
authors.initials: L
authors.lastname: Rodríguez
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Anna
authors.initials: A
authors.lastname: Solé
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Carolina
authors.initials: C
authors.lastname: Lliberós
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Núria
authors.initials: N
authors.lastname: Mencia
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Carlos J
authors.initials: CJ
authors.lastname: Ciudad
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Pharmacy, University of Barcelona , Barcelona, Spain .
authors.forename: Véronique
authors.initials: V
authors.lastname: Noé
mesh_terms: D017209:Apoptosis / Q000235:genetics; D020029:Base Pairing; D045744:Cell Line, Tumor; D002470:Cell Survival; D004264:DNA Topoisomerases, Type I / Q000235:genetics / Q000378:metabolism; D005260:Female; D020868:Gene Silencing; D052138:Genes, Neoplasm; D006801:Humans; D055029:Inverted Repeat Sequences; D008297:Male; D011071:Poly T / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: C000595707:BCL2 protein, human; C489427:MYC protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D011071:Poly T; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D004264:DNA Topoisomerases, Type I; C528909:TOP1 protein, human