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Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer

INTRODUCTION: Exosomes are 30-100 nm membrane vesicles of endocytic origin, mediating diverse biological functions including tumor cell invasion, cell-cell communication and antigen presentation through transfer of proteins, mRNAs and microRNAs. Recent evidence suggests that microRNAs can be release...

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Autores principales: Singh, Ramesh, Pochampally, Radhika, Watabe, Kounosuke, Lu, Zhaohui, Mo, Yin-Yuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4258287/
https://www.ncbi.nlm.nih.gov/pubmed/25428807
http://dx.doi.org/10.1186/1476-4598-13-256
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author Singh, Ramesh
Pochampally, Radhika
Watabe, Kounosuke
Lu, Zhaohui
Mo, Yin-Yuan
author_facet Singh, Ramesh
Pochampally, Radhika
Watabe, Kounosuke
Lu, Zhaohui
Mo, Yin-Yuan
author_sort Singh, Ramesh
collection PubMed
description INTRODUCTION: Exosomes are 30-100 nm membrane vesicles of endocytic origin, mediating diverse biological functions including tumor cell invasion, cell-cell communication and antigen presentation through transfer of proteins, mRNAs and microRNAs. Recent evidence suggests that microRNAs can be released through ceramide-dependent secretory machinery regulated by neutral sphingomyelinase 2 (nSMase2) enzyme encoded by the smpd3 gene that triggers exosome secretion. However, whether exosome-mediated microRNA transfer plays any role in cell invasion remains poorly understood. Thus, the aim of this study was to identify the exosomal microRNAs involved in breast cancer invasion. METHODS: The expression level of endogenous and exosomal miRNAs were examined by real time PCR and the expression level of target proteins were detected by western blot. Scanning electron and confocal microscopy were used to characterize exosomes and to study its uptake and transfer. Luciferase reporter plasmids and its mutant were used to confirm direct targeting. Furthermore, the functional significance of exosomal miR-10b was estimated by invasion assay. RESULTS: In this study, we demonstrate that microRNA carrying exosomes can be transferred among different cell lines through direct uptake. miR-10b is highly expressed in metastatic breast cancer MDA-MB-231 cells as compared to non-metastatic breast cancer cells or non-malignant breast cells; it is actively secreted into medium via exosomes. In particular, nSMase2 or ceramide promotes the exosome-mediated miR-10b secretion whereas ceramide inhibitor suppresses this secretion. Moreover, upon uptake, miR-10b can suppress the protein level of its target genes such as HOXD10 and KLF4, indicating its functional significance. Finally, treatment with exosomes derived from MDA-MB-231 cells could induce the invasion ability of non-malignant HMLE cells. CONCLUSION: Together, our results suggest that a set of specific microRNAs may play an important role in modulating tumor microenvironment through exosomes. Thus, a better understanding of this process may aid in the development of novel therapeutic agents. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1476-4598-13-256) contains supplementary material, which is available to authorized users.
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spelling pubmed-42582872014-12-08 Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer Singh, Ramesh Pochampally, Radhika Watabe, Kounosuke Lu, Zhaohui Mo, Yin-Yuan Mol Cancer Research INTRODUCTION: Exosomes are 30-100 nm membrane vesicles of endocytic origin, mediating diverse biological functions including tumor cell invasion, cell-cell communication and antigen presentation through transfer of proteins, mRNAs and microRNAs. Recent evidence suggests that microRNAs can be released through ceramide-dependent secretory machinery regulated by neutral sphingomyelinase 2 (nSMase2) enzyme encoded by the smpd3 gene that triggers exosome secretion. However, whether exosome-mediated microRNA transfer plays any role in cell invasion remains poorly understood. Thus, the aim of this study was to identify the exosomal microRNAs involved in breast cancer invasion. METHODS: The expression level of endogenous and exosomal miRNAs were examined by real time PCR and the expression level of target proteins were detected by western blot. Scanning electron and confocal microscopy were used to characterize exosomes and to study its uptake and transfer. Luciferase reporter plasmids and its mutant were used to confirm direct targeting. Furthermore, the functional significance of exosomal miR-10b was estimated by invasion assay. RESULTS: In this study, we demonstrate that microRNA carrying exosomes can be transferred among different cell lines through direct uptake. miR-10b is highly expressed in metastatic breast cancer MDA-MB-231 cells as compared to non-metastatic breast cancer cells or non-malignant breast cells; it is actively secreted into medium via exosomes. In particular, nSMase2 or ceramide promotes the exosome-mediated miR-10b secretion whereas ceramide inhibitor suppresses this secretion. Moreover, upon uptake, miR-10b can suppress the protein level of its target genes such as HOXD10 and KLF4, indicating its functional significance. Finally, treatment with exosomes derived from MDA-MB-231 cells could induce the invasion ability of non-malignant HMLE cells. CONCLUSION: Together, our results suggest that a set of specific microRNAs may play an important role in modulating tumor microenvironment through exosomes. Thus, a better understanding of this process may aid in the development of novel therapeutic agents. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1476-4598-13-256) contains supplementary material, which is available to authorized users. BioMed Central 2014-11-26 /pmc/articles/PMC4258287/ /pubmed/25428807 http://dx.doi.org/10.1186/1476-4598-13-256 Text en © Singh et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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
Singh, Ramesh
Pochampally, Radhika
Watabe, Kounosuke
Lu, Zhaohui
Mo, Yin-Yuan
Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title_full Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title_fullStr Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title_full_unstemmed Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title_short Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer
title_sort exosome-mediated transfer of mir-10b promotes cell invasion in breast cancer
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4258287/
https://www.ncbi.nlm.nih.gov/pubmed/25428807
http://dx.doi.org/10.1186/1476-4598-13-256
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