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 l...
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| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2017-01-01
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| Series: | Prostate Cancer |
| Online Access: | http://dx.doi.org/10.1155/2017/4893921 |
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| author | Jaroslaw Szczyrba Volker Jung Michaela Beitzinger Elke Nolte Sven Wach Martin Hart Sandra Sapich Marc Wiesehöfer Helge Taubert Gunther Wennemuth Norbert Eichner Thomas Stempfl Bernd Wullich Gunter Meister Friedrich A. Grässer |
| author_facet | Jaroslaw Szczyrba Volker Jung Michaela Beitzinger Elke Nolte Sven Wach Martin Hart Sandra Sapich Marc Wiesehöfer Helge Taubert Gunther Wennemuth Norbert Eichner Thomas Stempfl Bernd Wullich Gunter Meister Friedrich A. Grässer |
| author_sort | Jaroslaw Szczyrba |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-fa3d357e4fd848889507972bcd967529 |
| institution | Kabale University |
| issn | 2090-3111 2090-312X |
| language | English |
| publishDate | 2017-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Prostate Cancer |
| spelling | doaj-art-fa3d357e4fd848889507972bcd9675292025-02-03T05:58:55ZengWileyProstate Cancer2090-31112090-312X2017-01-01201710.1155/2017/48939214893921Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target GenesJaroslaw Szczyrba0Volker Jung1Michaela Beitzinger2Elke Nolte3Sven Wach4Martin Hart5Sandra Sapich6Marc Wiesehöfer7Helge Taubert8Gunther Wennemuth9Norbert Eichner10Thomas Stempfl11Bernd Wullich12Gunter Meister13Friedrich A. Grässer14Institute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, GermanyUniversity Clinic of Urology, Saarland University Medical School, Kirrbergerstrasse, 66421 Homburg/Saar, GermanyBiochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, GermanyUniversity Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, GermanyUniversity Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, GermanyInstitute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, GermanyInstitute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, GermanyInstitute of Anatomy, University Hospital Essen, Hufelandstrasse 55, 45147 Essen, GermanyUniversity Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, GermanyInstitute of Anatomy, University Hospital Essen, Hufelandstrasse 55, 45147 Essen, GermanyBiochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, GermanyCenter of Excellence for Fluorescent Bioanalytics (KFB), University of Regensburg, 93053 Regensburg, GermanyUniversity Clinic of Urology, Friedrich-Alexander-University Erlangen-Nürnberg, Krankenhausstrasse 12, 91054 Erlangen, GermanyBiochemistry Center Regensburg (BZR), Laboratory for RNA Biology, 93053 Regensburg, GermanyInstitute of Virology, Saarland University Medical School, Kirrbergerstrasse, Haus 47, 66421 Homburg/Saar, GermanyPosttranscriptional 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.http://dx.doi.org/10.1155/2017/4893921 |
| spellingShingle | Jaroslaw Szczyrba Volker Jung Michaela Beitzinger Elke Nolte Sven Wach Martin Hart Sandra Sapich Marc Wiesehöfer Helge Taubert Gunther Wennemuth Norbert Eichner Thomas Stempfl Bernd Wullich Gunter Meister Friedrich A. Grässer Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes Prostate Cancer |
| title | Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes |
| title_full | Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes |
| title_fullStr | Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes |
| title_full_unstemmed | Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes |
| title_short | Analysis of Argonaute Complex Bound mRNAs in DU145 Prostate Carcinoma Cells Reveals New miRNA Target Genes |
| title_sort | analysis of argonaute complex bound mrnas in du145 prostate carcinoma cells reveals new mirna target genes |
| url | http://dx.doi.org/10.1155/2017/4893921 |
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