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Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme

BACKGROUND: Literature reports that mature microRNA (miRNA) can be methylated at adenosine, guanosine and cytosine. However, the molecular mechanisms involved in cytosine methylation of miRNAs have not yet been fully elucidated. Here we investigated the biological role and underlying mechanism of cy...

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Autores principales: Cheray, Mathilde, Etcheverry, Amandine, Jacques, Camille, Pacaud, Romain, Bougras-Cartron, Gwenola, Aubry, Marc, Denoual, Florent, Peterlongo, Pierre, Nadaradjane, Arulraj, Briand, Joséphine, Akcha, Farida, Heymann, Dominique, Vallette, François M., Mosser, Jean, Ory, Benjamin, Cartron, Pierre-François
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041276/
https://www.ncbi.nlm.nih.gov/pubmed/32098627
http://dx.doi.org/10.1186/s12943-020-01155-z
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author Cheray, Mathilde
Etcheverry, Amandine
Jacques, Camille
Pacaud, Romain
Bougras-Cartron, Gwenola
Aubry, Marc
Denoual, Florent
Peterlongo, Pierre
Nadaradjane, Arulraj
Briand, Joséphine
Akcha, Farida
Heymann, Dominique
Vallette, François M.
Mosser, Jean
Ory, Benjamin
Cartron, Pierre-François
author_facet Cheray, Mathilde
Etcheverry, Amandine
Jacques, Camille
Pacaud, Romain
Bougras-Cartron, Gwenola
Aubry, Marc
Denoual, Florent
Peterlongo, Pierre
Nadaradjane, Arulraj
Briand, Joséphine
Akcha, Farida
Heymann, Dominique
Vallette, François M.
Mosser, Jean
Ory, Benjamin
Cartron, Pierre-François
author_sort Cheray, Mathilde
collection PubMed
description BACKGROUND: Literature reports that mature microRNA (miRNA) can be methylated at adenosine, guanosine and cytosine. However, the molecular mechanisms involved in cytosine methylation of miRNAs have not yet been fully elucidated. Here we investigated the biological role and underlying mechanism of cytosine methylation in miRNAs in glioblastoma multiforme (GBM). METHODS: RNA immunoprecipitation with the anti-5methylcytosine (5mC) antibody followed by Array, ELISA, dot blot, incorporation of a radio-labelled methyl group in miRNA, and miRNA bisulfite sequencing were perfomred to detect the cytosine methylation in mature miRNA. Cross-Linking immunoprecipiation qPCR, transfection with methylation/unmethylated mimic miRNA, luciferase promoter reporter plasmid, Biotin-tagged 3’UTR/mRNA or miRNA experiments and in vivo assays were used to investigate the role of methylated miRNAs. Finally, the prognostic value of methylated miRNAs was analyzed in a cohorte of GBM pateints. RESULTS: Our study reveals that a significant fraction of miRNAs contains 5mC. Cellular experiments show that DNMT3A/AGO4 methylated miRNAs at cytosine residues inhibit the formation of miRNA/mRNA duplex and leading to the loss of their repressive function towards gene expression. In vivo experiments show that cytosine-methylation of miRNA abolishes the tumor suppressor function of miRNA-181a-5p miRNA for example. Our study also reveals that cytosine-methylation of miRNA-181a-5p results is associated a poor prognosis in GBM patients. CONCLUSION: Together, our results indicate that the DNMT3A/AGO4-mediated cytosine methylation of miRNA negatively. GRAPHICAL ABSTRACT: [Image: see text]
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spelling pubmed-70412762020-03-03 Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme Cheray, Mathilde Etcheverry, Amandine Jacques, Camille Pacaud, Romain Bougras-Cartron, Gwenola Aubry, Marc Denoual, Florent Peterlongo, Pierre Nadaradjane, Arulraj Briand, Joséphine Akcha, Farida Heymann, Dominique Vallette, François M. Mosser, Jean Ory, Benjamin Cartron, Pierre-François Mol Cancer Research BACKGROUND: Literature reports that mature microRNA (miRNA) can be methylated at adenosine, guanosine and cytosine. However, the molecular mechanisms involved in cytosine methylation of miRNAs have not yet been fully elucidated. Here we investigated the biological role and underlying mechanism of cytosine methylation in miRNAs in glioblastoma multiforme (GBM). METHODS: RNA immunoprecipitation with the anti-5methylcytosine (5mC) antibody followed by Array, ELISA, dot blot, incorporation of a radio-labelled methyl group in miRNA, and miRNA bisulfite sequencing were perfomred to detect the cytosine methylation in mature miRNA. Cross-Linking immunoprecipiation qPCR, transfection with methylation/unmethylated mimic miRNA, luciferase promoter reporter plasmid, Biotin-tagged 3’UTR/mRNA or miRNA experiments and in vivo assays were used to investigate the role of methylated miRNAs. Finally, the prognostic value of methylated miRNAs was analyzed in a cohorte of GBM pateints. RESULTS: Our study reveals that a significant fraction of miRNAs contains 5mC. Cellular experiments show that DNMT3A/AGO4 methylated miRNAs at cytosine residues inhibit the formation of miRNA/mRNA duplex and leading to the loss of their repressive function towards gene expression. In vivo experiments show that cytosine-methylation of miRNA abolishes the tumor suppressor function of miRNA-181a-5p miRNA for example. Our study also reveals that cytosine-methylation of miRNA-181a-5p results is associated a poor prognosis in GBM patients. CONCLUSION: Together, our results indicate that the DNMT3A/AGO4-mediated cytosine methylation of miRNA negatively. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2020-02-25 /pmc/articles/PMC7041276/ /pubmed/32098627 http://dx.doi.org/10.1186/s12943-020-01155-z Text en © The Author(s) 2020 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Cheray, Mathilde
Etcheverry, Amandine
Jacques, Camille
Pacaud, Romain
Bougras-Cartron, Gwenola
Aubry, Marc
Denoual, Florent
Peterlongo, Pierre
Nadaradjane, Arulraj
Briand, Joséphine
Akcha, Farida
Heymann, Dominique
Vallette, François M.
Mosser, Jean
Ory, Benjamin
Cartron, Pierre-François
Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title_full Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title_fullStr Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title_full_unstemmed Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title_short Cytosine methylation of mature microRNAs inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
title_sort cytosine methylation of mature micrornas inhibits their functions and is associated with poor prognosis in glioblastoma multiforme
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041276/
https://www.ncbi.nlm.nih.gov/pubmed/32098627
http://dx.doi.org/10.1186/s12943-020-01155-z
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