Cargando…

The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9

BACKGROUND: MicroRNAs (miRNAs) are a group of endogenous small non-coding RNAs that regulate gene expression at the post-transcriptional level. A number of studies have led to the notion that some miRNAs have key roles in control of pancreatic islet development and insulin secretion. Based on some s...

Descripción completa

Detalles Bibliográficos
Autores principales: Jafarian, Arefeh, Taghikani, Mohammad, Abroun, Saeid, Allahverdi, Amir, Lamei, Maryam, Lakpour, Niknam, Soleimani, Masoud
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4457856/
https://www.ncbi.nlm.nih.gov/pubmed/26047014
http://dx.doi.org/10.1371/journal.pone.0128650
_version_ 1782375009313882112
author Jafarian, Arefeh
Taghikani, Mohammad
Abroun, Saeid
Allahverdi, Amir
Lamei, Maryam
Lakpour, Niknam
Soleimani, Masoud
author_facet Jafarian, Arefeh
Taghikani, Mohammad
Abroun, Saeid
Allahverdi, Amir
Lamei, Maryam
Lakpour, Niknam
Soleimani, Masoud
author_sort Jafarian, Arefeh
collection PubMed
description BACKGROUND: MicroRNAs (miRNAs) are a group of endogenous small non-coding RNAs that regulate gene expression at the post-transcriptional level. A number of studies have led to the notion that some miRNAs have key roles in control of pancreatic islet development and insulin secretion. Based on some studies on miRNAs pattern, the researchers in this paper investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. METHODOLOGY: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose so extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. Although derived IPCs by miR-375 alone were capable to express insulin and other endocrine specific transcription factors, the cells lacked the machinery to respond to glucose. CONCLUSION: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. Although the roles of miR-375 and miR-9 are well known in pancreatic development and insulin secretion, the use of these miRNAs in transdifferentiation was never demonstrated. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus.
format Online
Article
Text
id pubmed-4457856
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-44578562015-06-09 The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9 Jafarian, Arefeh Taghikani, Mohammad Abroun, Saeid Allahverdi, Amir Lamei, Maryam Lakpour, Niknam Soleimani, Masoud PLoS One Research Article BACKGROUND: MicroRNAs (miRNAs) are a group of endogenous small non-coding RNAs that regulate gene expression at the post-transcriptional level. A number of studies have led to the notion that some miRNAs have key roles in control of pancreatic islet development and insulin secretion. Based on some studies on miRNAs pattern, the researchers in this paper investigated the pancreatic differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) by up-regulation of miR-375 and down-regulation of miR-9 by lentiviruses containing miR-375 and anti-miR-9. METHODOLOGY: After 21 days of induction, islet-like clusters containing insulin producing cells (IPCs) were confirmed by dithizone (DTZ) staining. The IPCs and β cell specific related genes and proteins were detected using qRT-PCR and immunofluorescence on days 7, 14 and 21 of differentiation. Glucose challenge test was performed at different concentrations of glucose so extracellular and intracellular insulin and C-peptide were assayed using ELISA kit. Although derived IPCs by miR-375 alone were capable to express insulin and other endocrine specific transcription factors, the cells lacked the machinery to respond to glucose. CONCLUSION: It was found that over-expression of miR-375 led to a reduction in levels of Mtpn protein in derived IPCs, while treatment with anti-miR-9 following miR-375 over-expression had synergistic effects on MSCs differentiation and insulin secretion in a glucose-regulated manner. The researchers reported that silencing of miR-9 increased OC-2 protein in IPCs that may contribute to the observed glucose-regulated insulin secretion. Although the roles of miR-375 and miR-9 are well known in pancreatic development and insulin secretion, the use of these miRNAs in transdifferentiation was never demonstrated. These findings highlight miRNAs functions in stem cells differentiation and suggest that they could be used as therapeutic tools for gene-based therapy in diabetes mellitus. Public Library of Science 2015-06-05 /pmc/articles/PMC4457856/ /pubmed/26047014 http://dx.doi.org/10.1371/journal.pone.0128650 Text en © 2015 Jafarian et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Jafarian, Arefeh
Taghikani, Mohammad
Abroun, Saeid
Allahverdi, Amir
Lamei, Maryam
Lakpour, Niknam
Soleimani, Masoud
The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title_full The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title_fullStr The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title_full_unstemmed The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title_short The Generation of Insulin Producing Cells from Human Mesenchymal Stem Cells by MiR-375 and Anti-MiR-9
title_sort generation of insulin producing cells from human mesenchymal stem cells by mir-375 and anti-mir-9
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4457856/
https://www.ncbi.nlm.nih.gov/pubmed/26047014
http://dx.doi.org/10.1371/journal.pone.0128650
work_keys_str_mv AT jafarianarefeh thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT taghikanimohammad thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT abrounsaeid thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT allahverdiamir thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT lameimaryam thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT lakpourniknam thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT soleimanimasoud thegenerationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT jafarianarefeh generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT taghikanimohammad generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT abrounsaeid generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT allahverdiamir generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT lameimaryam generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT lakpourniknam generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9
AT soleimanimasoud generationofinsulinproducingcellsfromhumanmesenchymalstemcellsbymir375andantimir9