Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells

Detalhes bibliográficos
Autor(a) principal: Dong,Xia
Data de Publicação: 2011
Outros Autores: Song,Li-ping, Zhu,Dun-wan, Zhang,Hai-ling, Liu,Lan-xia, Leng,Xi-gang
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Genetics and Molecular Biology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000100005
Resumo: Tissue factor pathway inhibitor (TFPI) plays a vitally important role in the blood coagulation pathway. Recent studies indicated that TFPI induces apoptosis in vascular smooth-muscle cells (VSMCs) in animals. The present study investigated whether the TFPI gene could also induce apoptosis in human vascular smooth-muscle cells (hVSMCs). Such cells were isolated from human umbilical arteries and subsequently transfected with pIRES-TFPI plasmid (2 μg/mL). MTT assaying and cell counting were applied to measure cell viability and proliferation, RT-PCR was utilized to analyze TFPI gene expression in the cells. Apoptosis was analyzed by fluorescence activated cell sorting (FACS). Several key proteins involved in apoptosis were examined through Western blotting. It was shown that TFPI gene transfer led to its increased cellular expression, with a subsequent reduction in hVSMC proliferation. Further investigation demonstrated that TFPI gene expression resulted in lesser amounts of procaspase-3, procaspase-8 and procascase-9, and an increased release of mitochondrial cytochrome c (cyt-c) into cytoplasm, thereby implying the involvement of both extrinsic and intrinsic pathways in TFPI gene-induced apoptosis in hVSMCs.
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spelling Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cellstissue factor pathway inhibitorvascular smooth muscle cellsapoptosisTissue factor pathway inhibitor (TFPI) plays a vitally important role in the blood coagulation pathway. Recent studies indicated that TFPI induces apoptosis in vascular smooth-muscle cells (VSMCs) in animals. The present study investigated whether the TFPI gene could also induce apoptosis in human vascular smooth-muscle cells (hVSMCs). Such cells were isolated from human umbilical arteries and subsequently transfected with pIRES-TFPI plasmid (2 μg/mL). MTT assaying and cell counting were applied to measure cell viability and proliferation, RT-PCR was utilized to analyze TFPI gene expression in the cells. Apoptosis was analyzed by fluorescence activated cell sorting (FACS). Several key proteins involved in apoptosis were examined through Western blotting. It was shown that TFPI gene transfer led to its increased cellular expression, with a subsequent reduction in hVSMC proliferation. Further investigation demonstrated that TFPI gene expression resulted in lesser amounts of procaspase-3, procaspase-8 and procascase-9, and an increased release of mitochondrial cytochrome c (cyt-c) into cytoplasm, thereby implying the involvement of both extrinsic and intrinsic pathways in TFPI gene-induced apoptosis in hVSMCs.Sociedade Brasileira de Genética2011-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000100005Genetics and Molecular Biology v.34 n.1 2011reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/S1415-47572011000100005info:eu-repo/semantics/openAccessDong,XiaSong,Li-pingZhu,Dun-wanZhang,Hai-lingLiu,Lan-xiaLeng,Xi-gangeng2011-01-28T00:00:00Zoai:scielo:S1415-47572011000100005Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2011-01-28T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false
dc.title.none.fl_str_mv Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
title Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
spellingShingle Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
Dong,Xia
tissue factor pathway inhibitor
vascular smooth muscle cells
apoptosis
title_short Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
title_full Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
title_fullStr Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
title_full_unstemmed Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
title_sort Impact of the tissue factor pathway inhibitor gene on apoptosis in human vascular smooth muscle cells
author Dong,Xia
author_facet Dong,Xia
Song,Li-ping
Zhu,Dun-wan
Zhang,Hai-ling
Liu,Lan-xia
Leng,Xi-gang
author_role author
author2 Song,Li-ping
Zhu,Dun-wan
Zhang,Hai-ling
Liu,Lan-xia
Leng,Xi-gang
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Dong,Xia
Song,Li-ping
Zhu,Dun-wan
Zhang,Hai-ling
Liu,Lan-xia
Leng,Xi-gang
dc.subject.por.fl_str_mv tissue factor pathway inhibitor
vascular smooth muscle cells
apoptosis
topic tissue factor pathway inhibitor
vascular smooth muscle cells
apoptosis
description Tissue factor pathway inhibitor (TFPI) plays a vitally important role in the blood coagulation pathway. Recent studies indicated that TFPI induces apoptosis in vascular smooth-muscle cells (VSMCs) in animals. The present study investigated whether the TFPI gene could also induce apoptosis in human vascular smooth-muscle cells (hVSMCs). Such cells were isolated from human umbilical arteries and subsequently transfected with pIRES-TFPI plasmid (2 μg/mL). MTT assaying and cell counting were applied to measure cell viability and proliferation, RT-PCR was utilized to analyze TFPI gene expression in the cells. Apoptosis was analyzed by fluorescence activated cell sorting (FACS). Several key proteins involved in apoptosis were examined through Western blotting. It was shown that TFPI gene transfer led to its increased cellular expression, with a subsequent reduction in hVSMC proliferation. Further investigation demonstrated that TFPI gene expression resulted in lesser amounts of procaspase-3, procaspase-8 and procascase-9, and an increased release of mitochondrial cytochrome c (cyt-c) into cytoplasm, thereby implying the involvement of both extrinsic and intrinsic pathways in TFPI gene-induced apoptosis in hVSMCs.
publishDate 2011
dc.date.none.fl_str_mv 2011-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
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status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000100005
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000100005
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1415-47572011000100005
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Sociedade Brasileira de Genética
publisher.none.fl_str_mv Sociedade Brasileira de Genética
dc.source.none.fl_str_mv Genetics and Molecular Biology v.34 n.1 2011
reponame:Genetics and Molecular Biology
instname:Sociedade Brasileira de Genética (SBG)
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instname_str Sociedade Brasileira de Genética (SBG)
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