title: Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis.
abstract: Of the over 200 identified mammalian microRNAs (miRNAs), only a few have known biological activity. To gain a better understanding of the role that miRNAs play in specific cellular pathways, we utilized antisense molecules to inhibit miRNA activity. We used miRNA inhibitors targeting miR-23, 21, 15a, 16 and 19a to test efficacy of antisense molecules in reducing miRNA activity on reporter genes bearing miRNA-binding sites. The miRNA inhibitors de-repressed reporter gene activity when a miRNA-binding site was cloned into its 3'-untranslated region. We employed a library of miRNA inhibitors to screen for miRNA involved in cell growth and apoptosis. In HeLa cells, we found that inhibition of miR-95, 124, 125, 133, 134, 144, 150, 152, 187, 190, 191, 192, 193, 204, 211, 218, 220, 296 and 299 caused a decrease in cell growth and that inhibition of miR-21 and miR-24 had a profound increase in cell growth. On the other hand, inhibition of miR-7, 19a, 23, 24, 134, 140, 150, 192 and 193 down-regulated cell growth, and miR-107, 132, 155, 181, 191, 194, 203, 215 and 301 increased cell growth in lung carcinoma cells, A549. We also identified miRNA that when inhibited increased the level of apoptosis (miR-1d, 7, 148, 204, 210, 216 and 296) and one miRNA that decreased apoptosis (miR-214) in HeLa cells. From these screens, we conclude that miRNA-mediated regulation has a complexity of cellular outcomes and that miRNAs can be mediators of regulation of cell growth and apoptosis pathways.
authors.affiliation: Ambion, Inc. 2130 Woodward Street, Austin, TX 78744-1832, USA.
authors.forename: Angie M
authors.initials: AM
authors.lastname: Cheng
authors.forename: Mike W
authors.initials: MW
authors.lastname: Byrom
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Shelton
authors.forename: Lance P
authors.initials: LP
authors.lastname: Ford
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*
chemical_list: D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D008156:Luciferases
references.citation: Cell. 1993 Dec 3;75(5):855-62
references.pmid: 8252622
references.citation: Cell. 1993 Dec 3;75(5):843-54
references.pmid: 8252621
references.citation: Mol Cell. 2000 Apr;5(4):659-69
references.pmid: 10882102
references.citation: Cell. 2001 Dec 28;107(7):823-6
references.pmid: 11779458
references.citation: Mol Cell. 2002 Jun;9(6):1327-33
references.pmid: 12086629
references.citation: EMBO J. 2002 Sep 2;21(17):4663-70
references.pmid: 12198168
references.citation: Science. 2002 Sep 20;297(5589):2056-60
references.pmid: 12154197
references.citation: Annu Rev Cell Dev Biol. 2002;18:495-513
references.pmid: 12142272
references.citation: Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15524-9
references.pmid: 12434020
references.citation: Cell. 2003 Apr 4;113(1):25-36
references.pmid: 12679032
references.citation: Curr Biol. 2003 Apr 29;13(9):790-5
references.pmid: 12725740
references.citation: Proc Natl Acad Sci U S A. 2003 Aug 19;100(17):9779-84
references.pmid: 12902540
references.citation: Nucleic Acids Res. 2003 Sep 1;31(17):4973-80
references.pmid: 12930946
references.citation: Nature. 2003 Sep 25;425(6956):415-9
references.pmid: 14508493
references.citation: Mol Cancer Res. 2003 Oct;1(12):882-91
references.pmid: 14573789
references.citation: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D109-11
references.pmid: 14681370
references.citation: Genes Chromosomes Cancer. 2004 Feb;39(2):167-9
references.pmid: 14695998
references.citation: Cell. 2003 Dec 26;115(7):787-98
references.pmid: 14697198
references.citation: Genes Dev. 2003 Dec 15;17(24):3011-6
references.pmid: 14681208
references.citation: Science. 2004 Jan 2;303(5654):83-6
references.pmid: 14657504
references.citation: Genome Biol. 2003;5(1):R1
references.pmid: 14709173
references.citation: Cell. 2004 Jan 23;116(2):281-97
references.pmid: 14744438
references.citation: RNA. 2004 Mar;10(3):544-50
references.pmid: 14970398
references.citation: Genes Dev. 2004 Mar 1;18(5):504-11
references.pmid: 15014042
references.citation: PLoS Biol. 2004 Apr;2(4):E98
references.pmid: 15024405
references.citation: Science. 2004 Apr 23;304(5670):594-6
references.pmid: 15105502
references.citation: Nature. 2000 Feb 24;403(6772):901-6
references.pmid: 10706289