Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | International Braz J Urol (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382015000400764 |
Resumo: | ABSTRACTPurpose:RNA activation (RNAa) is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs), also known as small activating RNAs (saRNAs). Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI) is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs) into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD) into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI)–a condition primarily resulted from urethral sphincter deficiency. |
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International Braz J Urol (Online) |
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Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoDRNAMyoD ProteinUrinary Bladder NeoplasmsUrinary IncontinenceStressDesminABSTRACTPurpose:RNA activation (RNAa) is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs), also known as small activating RNAs (saRNAs). Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI) is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs) into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD) into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI)–a condition primarily resulted from urethral sphincter deficiency.Sociedade Brasileira de Urologia2015-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382015000400764International braz j urol v.41 n.4 2015reponame:International Braz J Urol (Online)instname:Sociedade Brasileira de Urologia (SBU)instacron:SBU10.1590/S1677-5538.IBJU.2014.0400info:eu-repo/semantics/openAccessWang,ChengheChen,ZhongWu,JiaZhang,YanHu,JiaGe,QiangqiangWang,TaoYang,WeiminXu,HuaLiu,JihongYe,Zhangquneng2015-10-13T00:00:00Zoai:scielo:S1677-55382015000400764Revistahttp://www.brazjurol.com.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||brazjurol@brazjurol.com.br1677-61191677-5538opendoar:2015-10-13T00:00International Braz J Urol (Online) - Sociedade Brasileira de Urologia (SBU)false |
dc.title.none.fl_str_mv |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
title |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
spellingShingle |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD Wang,Chenghe RNA MyoD Protein Urinary Bladder Neoplasms Urinary Incontinence Stress Desmin |
title_short |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
title_full |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
title_fullStr |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
title_full_unstemmed |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
title_sort |
Small activating RNA induces myogenic differentiation of rat adipose-derived stem cells by upregulating MyoD |
author |
Wang,Chenghe |
author_facet |
Wang,Chenghe Chen,Zhong Wu,Jia Zhang,Yan Hu,Jia Ge,Qiangqiang Wang,Tao Yang,Weimin Xu,Hua Liu,Jihong Ye,Zhangqun |
author_role |
author |
author2 |
Chen,Zhong Wu,Jia Zhang,Yan Hu,Jia Ge,Qiangqiang Wang,Tao Yang,Weimin Xu,Hua Liu,Jihong Ye,Zhangqun |
author2_role |
author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Wang,Chenghe Chen,Zhong Wu,Jia Zhang,Yan Hu,Jia Ge,Qiangqiang Wang,Tao Yang,Weimin Xu,Hua Liu,Jihong Ye,Zhangqun |
dc.subject.por.fl_str_mv |
RNA MyoD Protein Urinary Bladder Neoplasms Urinary Incontinence Stress Desmin |
topic |
RNA MyoD Protein Urinary Bladder Neoplasms Urinary Incontinence Stress Desmin |
description |
ABSTRACTPurpose:RNA activation (RNAa) is a mechanism of gene activation triggered by promoter-targeted small double stranded RNAs (dsRNAs), also known as small activating RNAs (saRNAs). Myogenic regulatory factor MyoD is regarded as the master activator of myogenic differentiation cascade by binding to enhancer of muscle specific genes. Stress urinary incontinence (SUI) is a condition primarily resulted from urethral sphincter deficiency. It is thus expected that by promoting differentiation of adipose-derived stem cells (ADSCs) into myoblasts by activating MyoD gene through RNAa may offer benefits to SUI.Materials and Methods:Rats ADSCs were isolated, proliferated in vitro, and identified by flow cytometry. Purified ADSCs were then transfected with a MyoD saRNA or control transfected. Real-time polymerase chain reaction (RT-PCR) and western blotting were used to detect MyoD mRNA and protein expression, respectively. Immunocytochemical staining was applied to determine the expression of desmin protein in transfected cells. Cell viability was measured by using CellTiter 96® AQueous One Solution Cell Proliferation Assay kit.Results:Transfection of a MyoD saRNA (dsMyoD) into ADSCs significantly induced the expression of MyoD at both the mRNA and protein levels, and inhibited cell proliferation. Desmin protein expression was detected in dsMyoD treated ADSCs 2 weeks later.Conclusion:Our findings show that RNAa mediated overexpression of MyoD can promote transdifferentiation of ADSCs into myoblasts and may help treat stress urinary incontinence (SUI)–a condition primarily resulted from urethral sphincter deficiency. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-08-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382015000400764 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382015000400764 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1677-5538.IBJU.2014.0400 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Urologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Urologia |
dc.source.none.fl_str_mv |
International braz j urol v.41 n.4 2015 reponame:International Braz J Urol (Online) instname:Sociedade Brasileira de Urologia (SBU) instacron:SBU |
instname_str |
Sociedade Brasileira de Urologia (SBU) |
instacron_str |
SBU |
institution |
SBU |
reponame_str |
International Braz J Urol (Online) |
collection |
International Braz J Urol (Online) |
repository.name.fl_str_mv |
International Braz J Urol (Online) - Sociedade Brasileira de Urologia (SBU) |
repository.mail.fl_str_mv |
||brazjurol@brazjurol.com.br |
_version_ |
1750318074453557248 |