MicroRNA-21 (miR-21) is a key regulator of oncogenic processes. It is significantly elevated in the majority of human tumors and functionally linked to cellular proliferation, survival and migration. In this study, we used two experimental-based strategies to search for novel miR-21 targets. On the one hand, we performed a proteomic approach using two-dimensional differential gel electrophoresis (2D-DIGE) to identify proteins suppressed upon enhanced miR-21 expression in LNCaP human prostate carcinoma cells. The tumor suppressor acidic nuclear phosphoprotein 32 family, member A (ANP32A) (alias pp32 or LANP) emerged as the most strongly downregulated protein. On the other hand, we applied a mathematical approach to select correlated gene sets that are negatively correlated with primary-miR-21 (pri-miR-21) expression in published transcriptome data from 114 B-cell lymphoma cases. Among these candidates, we found tumor suppressor SMARCA4 (alias BRG1) together with the already validated miR-21 target, PDCD4. ANP32A and SMARCA4, which are both involved in chromatin remodeling processes, were confirmed as direct miR-21 targets by immunoblot analysis and reporter gene assays. Furthermore, knock down of ANP32A mimicked the effect of enforced miR-21 expression by enhancing LNCaP cell viability, whereas overexpression of ANP32A in the presence of high miR-21 levels abrogated the miR-21-mediated effect. In A172 glioblastoma cells, enhanced ANP32A expression compensated for the effects of anti-miR-21 treatment on cell viability and apoptosis. In addition, miR-21 expression clearly increased the invasiveness of LNCaP cells, an effect also seen in part upon downregulation of ANP32A. In conclusion, these results suggest that downregulation of ANP32A contributes to the oncogenic function of miR-21.
%0 Journal Article
%1 Schramedei.2011
%A Schramedei, K.
%A Mörbt, N.
%A Pfeifer, G.
%A Läuter, J.
%A Rosolowski, M.
%A Tomm, J. M.
%A Bergen, M. von
%A Horn, F.
%A Brocke-Heidrich, K.
%D 2011
%J Oncogene
%K Base_Sequence Cell_Survival/genetics Cell_Transformation,_Neoplastic/genetics Cells,_Cultured DNA_Helicases/genetics/metabolism Down-Regulation Gene_Expression_Regulation Gene_Targeting Genes,_Tumor_Suppressor Humans Intracellular_Signaling_Peptides_and_Proteins/genetics/metabolism MicroRNAs/genetics/metabolism/physiology Molecular_Sequence_Data Nuclear_Proteins/genetics/metabolism Regulatory_Sequences,_Ribonucleic_Acid/genetics/physiology Sequence_Homology,_Nucleic_Acid Transcription_Factors/genetics/metabolism Two-Dimensional_Difference_Gel_Electrophoresis
%N 26
%P 2975–2985
%T MicroRNA-21 targets tumor suppressor genes ANP32A and SMARCA4
%V 30
%X MicroRNA-21 (miR-21) is a key regulator of oncogenic processes. It is significantly elevated in the majority of human tumors and functionally linked to cellular proliferation, survival and migration. In this study, we used two experimental-based strategies to search for novel miR-21 targets. On the one hand, we performed a proteomic approach using two-dimensional differential gel electrophoresis (2D-DIGE) to identify proteins suppressed upon enhanced miR-21 expression in LNCaP human prostate carcinoma cells. The tumor suppressor acidic nuclear phosphoprotein 32 family, member A (ANP32A) (alias pp32 or LANP) emerged as the most strongly downregulated protein. On the other hand, we applied a mathematical approach to select correlated gene sets that are negatively correlated with primary-miR-21 (pri-miR-21) expression in published transcriptome data from 114 B-cell lymphoma cases. Among these candidates, we found tumor suppressor SMARCA4 (alias BRG1) together with the already validated miR-21 target, PDCD4. ANP32A and SMARCA4, which are both involved in chromatin remodeling processes, were confirmed as direct miR-21 targets by immunoblot analysis and reporter gene assays. Furthermore, knock down of ANP32A mimicked the effect of enforced miR-21 expression by enhancing LNCaP cell viability, whereas overexpression of ANP32A in the presence of high miR-21 levels abrogated the miR-21-mediated effect. In A172 glioblastoma cells, enhanced ANP32A expression compensated for the effects of anti-miR-21 treatment on cell viability and apoptosis. In addition, miR-21 expression clearly increased the invasiveness of LNCaP cells, an effect also seen in part upon downregulation of ANP32A. In conclusion, these results suggest that downregulation of ANP32A contributes to the oncogenic function of miR-21.
@article{Schramedei.2011,
abstract = {MicroRNA-21 (miR-21) is a key regulator of oncogenic processes. It is significantly elevated in the majority of human tumors and functionally linked to cellular proliferation, survival and migration. In this study, we used two experimental-based strategies to search for novel miR-21 targets. On the one hand, we performed a proteomic approach using two-dimensional differential gel electrophoresis (2D-DIGE) to identify proteins suppressed upon enhanced miR-21 expression in LNCaP human prostate carcinoma cells. The tumor suppressor acidic nuclear phosphoprotein 32 family, member A (ANP32A) (alias pp32 or LANP) emerged as the most strongly downregulated protein. On the other hand, we applied a mathematical approach to select correlated gene sets that are negatively correlated with primary-miR-21 (pri-miR-21) expression in published transcriptome data from 114 B-cell lymphoma cases. Among these candidates, we found tumor suppressor SMARCA4 (alias BRG1) together with the already validated miR-21 target, PDCD4. ANP32A and SMARCA4, which are both involved in chromatin remodeling processes, were confirmed as direct miR-21 targets by immunoblot analysis and reporter gene assays. Furthermore, knock down of ANP32A mimicked the effect of enforced miR-21 expression by enhancing LNCaP cell viability, whereas overexpression of ANP32A in the presence of high miR-21 levels abrogated the miR-21-mediated effect. In A172 glioblastoma cells, enhanced ANP32A expression compensated for the effects of anti-miR-21 treatment on cell viability and apoptosis. In addition, miR-21 expression clearly increased the invasiveness of LNCaP cells, an effect also seen in part upon downregulation of ANP32A. In conclusion, these results suggest that downregulation of ANP32A contributes to the oncogenic function of miR-21.},
added-at = {2014-10-14T15:28:12.000+0200},
author = {Schramedei, K. and Mörbt, N. and Pfeifer, G. and Läuter, J. and Rosolowski, M. and Tomm, J. M. and Bergen, M. von and Horn, F. and Brocke-Heidrich, K.},
biburl = {https://www.bibsonomy.org/bibtex/2bb6dab1212efb90739d536c4076019f6/drtester},
interhash = {861d8d5b625ae01011f1a4f637b70619},
intrahash = {bb6dab1212efb90739d536c4076019f6},
journal = {Oncogene},
keywords = {Base_Sequence Cell_Survival/genetics Cell_Transformation,_Neoplastic/genetics Cells,_Cultured DNA_Helicases/genetics/metabolism Down-Regulation Gene_Expression_Regulation Gene_Targeting Genes,_Tumor_Suppressor Humans Intracellular_Signaling_Peptides_and_Proteins/genetics/metabolism MicroRNAs/genetics/metabolism/physiology Molecular_Sequence_Data Nuclear_Proteins/genetics/metabolism Regulatory_Sequences,_Ribonucleic_Acid/genetics/physiology Sequence_Homology,_Nucleic_Acid Transcription_Factors/genetics/metabolism Two-Dimensional_Difference_Gel_Electrophoresis},
number = 26,
pages = {2975–2985},
timestamp = {2014-10-14T15:28:12.000+0200},
title = {MicroRNA-21 targets tumor suppressor genes ANP32A and SMARCA4},
volume = 30,
year = 2011
}