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miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death

BACKGROUND: Acute myeloid leukaemia (AML) with nucleophosmin-1 (NPM1) mutation is a major subtype of AML. The NPM1 mutation induces a myeloproliferative disorder, but evidence indicates that other insults are necessary for the development of AML. We utilised microRNA microarrays and functional assay...

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Autores principales: Bryant, Adam, Palma, Catalina A, Jayaswal, Vivek, Yang, Yee Wa, Lutherborrow, Mark, Ma, David DF
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306826/
https://www.ncbi.nlm.nih.gov/pubmed/22348345
http://dx.doi.org/10.1186/1476-4598-11-8
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author Bryant, Adam
Palma, Catalina A
Jayaswal, Vivek
Yang, Yee Wa
Lutherborrow, Mark
Ma, David DF
author_facet Bryant, Adam
Palma, Catalina A
Jayaswal, Vivek
Yang, Yee Wa
Lutherborrow, Mark
Ma, David DF
author_sort Bryant, Adam
collection PubMed
description BACKGROUND: Acute myeloid leukaemia (AML) with nucleophosmin-1 (NPM1) mutation is a major subtype of AML. The NPM1 mutation induces a myeloproliferative disorder, but evidence indicates that other insults are necessary for the development of AML. We utilised microRNA microarrays and functional assays to determine if microRNA dysregulation could be involved in the pathogenesis of in NPM1 mutated (NPM1(mut))-AML. RESULTS: We used a stringent locked nucleic acid (LNA) based microRNA microarray platform to profile bone marrow samples of patients with normal karyotype AML. A panel of five microRNAs dichotomised AML patients according to their NPM1 mutational status. miR-10a, let-7b and let-7c were significantly over-expressed, while miR-130a and miR-335 were under-expressed in NPM1(mut)-AML when compared to NPM1(wildtype)-AML. Of these, miR-10a is the most differentially expressed in NPM1(mut)-AML versus NPM1(wildtype)-AML (> 10 fold higher as confirmed by qRT-PCR). To investigate the functions of miR-10a, the OCI-AML3 cell line was utilised, which is the only commercially available cell line bearing NPM1(mut). OCI-AML3 cells were firstly demonstrated to have a similarly high miR-10a expression to primary NPM1(mut)-AML patient samples. Inhibition of miR-10a expression by miRCURY LNA Inhibitors (Exiqon) in these cells resulted in increased cell death as assessed by MTS, cell cycle and Annexin-V assays and reduced clonogenic capacity, indicative of an involvement in leukaemic cell survival. In silico filtering of bioinformatically predicted targets of miR-10a identified a number of potential mRNA targets with annotated functions in haematopoiesis, cell growth and apoptosis. Lucferase reporter assays confirmed a number of these putative tumorogenic genes that are miR-10a suppressible including KLF4 and RB1CC1. This provides a potential mechanism for the pathogenic role of miR-10a in NPM1(mut)-AML. CONCLUSIONS: This study provides, for the first time, in vitro evidence of a pro-survival role of miR-10a in NPM1(mut)-AML, that it may contribute to the pathogenesis of NPM1(mut)-AML and identifies putative tumorogenic targets.
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spelling pubmed-33068262012-03-19 miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death Bryant, Adam Palma, Catalina A Jayaswal, Vivek Yang, Yee Wa Lutherborrow, Mark Ma, David DF Mol Cancer Research BACKGROUND: Acute myeloid leukaemia (AML) with nucleophosmin-1 (NPM1) mutation is a major subtype of AML. The NPM1 mutation induces a myeloproliferative disorder, but evidence indicates that other insults are necessary for the development of AML. We utilised microRNA microarrays and functional assays to determine if microRNA dysregulation could be involved in the pathogenesis of in NPM1 mutated (NPM1(mut))-AML. RESULTS: We used a stringent locked nucleic acid (LNA) based microRNA microarray platform to profile bone marrow samples of patients with normal karyotype AML. A panel of five microRNAs dichotomised AML patients according to their NPM1 mutational status. miR-10a, let-7b and let-7c were significantly over-expressed, while miR-130a and miR-335 were under-expressed in NPM1(mut)-AML when compared to NPM1(wildtype)-AML. Of these, miR-10a is the most differentially expressed in NPM1(mut)-AML versus NPM1(wildtype)-AML (> 10 fold higher as confirmed by qRT-PCR). To investigate the functions of miR-10a, the OCI-AML3 cell line was utilised, which is the only commercially available cell line bearing NPM1(mut). OCI-AML3 cells were firstly demonstrated to have a similarly high miR-10a expression to primary NPM1(mut)-AML patient samples. Inhibition of miR-10a expression by miRCURY LNA Inhibitors (Exiqon) in these cells resulted in increased cell death as assessed by MTS, cell cycle and Annexin-V assays and reduced clonogenic capacity, indicative of an involvement in leukaemic cell survival. In silico filtering of bioinformatically predicted targets of miR-10a identified a number of potential mRNA targets with annotated functions in haematopoiesis, cell growth and apoptosis. Lucferase reporter assays confirmed a number of these putative tumorogenic genes that are miR-10a suppressible including KLF4 and RB1CC1. This provides a potential mechanism for the pathogenic role of miR-10a in NPM1(mut)-AML. CONCLUSIONS: This study provides, for the first time, in vitro evidence of a pro-survival role of miR-10a in NPM1(mut)-AML, that it may contribute to the pathogenesis of NPM1(mut)-AML and identifies putative tumorogenic targets. BioMed Central 2012-02-20 /pmc/articles/PMC3306826/ /pubmed/22348345 http://dx.doi.org/10.1186/1476-4598-11-8 Text en Copyright ©2012 Bryant et al; BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Bryant, Adam
Palma, Catalina A
Jayaswal, Vivek
Yang, Yee Wa
Lutherborrow, Mark
Ma, David DF
miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title_full miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title_fullStr miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title_full_unstemmed miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title_short miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
title_sort mir-10a is aberrantly overexpressed in nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306826/
https://www.ncbi.nlm.nih.gov/pubmed/22348345
http://dx.doi.org/10.1186/1476-4598-11-8
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