Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes.
Posttranscriptional gene regulation by microRNAs (miRNAs) contributes to the induction and maintenance of prostate carcinoma (PCa). To identify mRNAs enriched or removed from Ago2-containing RISC complexes, these complexes were immunoprecipitated from normal prostate fibroblasts (PNFs) and the PCa line DU145 and the bound mRNAs were quantified by microarray. The analysis of Ago complexes derived from PNFs or DU145 confirmed the enrichment or depletion of a variety of mRNAs already known from the literature to be deregulated. Novel potential targets were analyzed by luciferase assays with miRNAs known to be deregulated in PCa. We demonstrate that the mRNAs of the death effector domain-containing protein (DEDD), the tumor necrosis factor receptor superfamily, member 10b protein (TNFRSF10B), the tumor protein p53 inducible nuclear protein 1 (TP53INP1), and the secreted protein, acidic, cysteine-rich (SPARC; osteonectin) are regulated by miRNAs miR-148a, miR-20a, miR-24, and miR-29a/b, respectively. Therefore, these miRNAs represent potential targets for therapy. Surprisingly, overexpression of miR-24 induced focus formation and proliferation of DU145 cells, while miR-29b reduced proliferation. The study confirms genes deregulated in PCa by virtue of their presence/absence in the Ago2-complex. In conjunction with the already published miRNA profiles of PCa, the data can be used to identify miRNA-regulated mRNAs.
Institute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, Germany; Institute of Anatomy, University Hospital Essen, Hufelandstrasse 55, 45147 Essen, Germany.
Jaroslaw
0000-0002-5950-0742
J
Szczyrba
University Clinic of Urology, Saarland University Medical School, Kirrbergerstrasse, 66421 Homburg/Saar, Germany.
Volker
V
Jung
Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, Germany.
Michaela
M
Beitzinger
University Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, Germany.
Elke
E
Nolte
University Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, Germany.
Sven
S
Wach
Institute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, Germany.
Martin
M
Hart
Institute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, Germany.
Sandra
S
Sapich
Institute of Anatomy, University Hospital Essen, Hufelandstrasse 55, 45147 Essen, Germany.
Marc
M
Wiesehöfer
University Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, Germany.
Helge
0000-0002-9077-1727
H
Taubert
Institute of Anatomy, University Hospital Essen, Hufelandstrasse 55, 45147 Essen, Germany.
Gunther
G
Wennemuth
Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, Germany.
Norbert
N
Eichner
Center of Excellence for Fluorescent Bioanalytics (KFB), University of Regensburg, 93053 Regensburg, Germany.
Thomas
T
Stempfl
University Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, Germany.
Bernd
B
Wullich
Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, Germany.
Gunter
G
Meister
Institute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, Germany.
Friedrich A
0000-0001-6234-5571
FA
Grässer