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Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance

MicroRNAs (miRNAs) have been widely studied in order to elucidate their biological functions. MicroRNA microarrays or miRNA overexpression libraries generated by synthesis and cloning of individual miRNAs have been used to study their different roles. In this work, we have developed a novel methodol...

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Autores principales: Moreno-Mateos, Miguel A., Barragán, Verónica, Torres, Belén, Rodríguez-Mateo, Cristina, Méndez-Vidal, Cristina, Berezikov, Eugene, Mudduluru, Giridhar, Allgayer, Heike, Pintor-Toro, José A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3884670/
https://www.ncbi.nlm.nih.gov/pubmed/24129493
http://dx.doi.org/10.1261/rna.039461.113
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author Moreno-Mateos, Miguel A.
Barragán, Verónica
Torres, Belén
Rodríguez-Mateo, Cristina
Méndez-Vidal, Cristina
Berezikov, Eugene
Mudduluru, Giridhar
Allgayer, Heike
Pintor-Toro, José A.
author_facet Moreno-Mateos, Miguel A.
Barragán, Verónica
Torres, Belén
Rodríguez-Mateo, Cristina
Méndez-Vidal, Cristina
Berezikov, Eugene
Mudduluru, Giridhar
Allgayer, Heike
Pintor-Toro, José A.
author_sort Moreno-Mateos, Miguel A.
collection PubMed
description MicroRNAs (miRNAs) have been widely studied in order to elucidate their biological functions. MicroRNA microarrays or miRNA overexpression libraries generated by synthesis and cloning of individual miRNAs have been used to study their different roles. In this work, we have developed a novel methodology to express mature miRNAs and other small RNAs from a double convergent RNA polymerase III promoter. We show that the generated miRNAs function similarly to those processed from primary transcripts or pri-miRNAs. This system allowed us to produce a lentiviral library expressing the whole population of small RNAs present in a metastatic cell line. A functional screening using this library led to the identification of hsa-miR-30b and hsa-miR-30c as negative regulators of cell death induced by loss of attachment (anoikis). Importantly, we demonstrated that the acquisition of anoikis resistance via these miRNAs is achieved through down-regulation of caspase 3 expression. Moreover, overexpression of these miRNAs resulted in a decrease of other types of caspase 3-dependent cell death and enhanced the survival of MCF10A acinar cells in morphogenesis assays, suggesting a putative role as oncomirs. In summary, this novel methodology provides a powerful and effective way for identifying novel small RNAs involved in a particular biological process.
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spelling pubmed-38846702014-12-01 Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance Moreno-Mateos, Miguel A. Barragán, Verónica Torres, Belén Rodríguez-Mateo, Cristina Méndez-Vidal, Cristina Berezikov, Eugene Mudduluru, Giridhar Allgayer, Heike Pintor-Toro, José A. RNA Articles MicroRNAs (miRNAs) have been widely studied in order to elucidate their biological functions. MicroRNA microarrays or miRNA overexpression libraries generated by synthesis and cloning of individual miRNAs have been used to study their different roles. In this work, we have developed a novel methodology to express mature miRNAs and other small RNAs from a double convergent RNA polymerase III promoter. We show that the generated miRNAs function similarly to those processed from primary transcripts or pri-miRNAs. This system allowed us to produce a lentiviral library expressing the whole population of small RNAs present in a metastatic cell line. A functional screening using this library led to the identification of hsa-miR-30b and hsa-miR-30c as negative regulators of cell death induced by loss of attachment (anoikis). Importantly, we demonstrated that the acquisition of anoikis resistance via these miRNAs is achieved through down-regulation of caspase 3 expression. Moreover, overexpression of these miRNAs resulted in a decrease of other types of caspase 3-dependent cell death and enhanced the survival of MCF10A acinar cells in morphogenesis assays, suggesting a putative role as oncomirs. In summary, this novel methodology provides a powerful and effective way for identifying novel small RNAs involved in a particular biological process. Cold Spring Harbor Laboratory Press 2013-12 /pmc/articles/PMC3884670/ /pubmed/24129493 http://dx.doi.org/10.1261/rna.039461.113 Text en © 2013 Moreno-Mateos et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.
spellingShingle Articles
Moreno-Mateos, Miguel A.
Barragán, Verónica
Torres, Belén
Rodríguez-Mateo, Cristina
Méndez-Vidal, Cristina
Berezikov, Eugene
Mudduluru, Giridhar
Allgayer, Heike
Pintor-Toro, José A.
Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title_full Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title_fullStr Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title_full_unstemmed Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title_short Novel small RNA expression libraries uncover hsa-miR-30b and hsa-miR-30c as important factors in anoikis resistance
title_sort novel small rna expression libraries uncover hsa-mir-30b and hsa-mir-30c as important factors in anoikis resistance
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3884670/
https://www.ncbi.nlm.nih.gov/pubmed/24129493
http://dx.doi.org/10.1261/rna.039461.113
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