PGC-1α辅助激活人PEPCK基因转录

【目的】构建人磷酸烯醇式丙酮酸羧激酶(PEPCK)基因启动子荧光素酶报告质粒PGL3-hPCK-luc，并探讨过氧化物酶体增殖物活化受体γ辅助激活因子1α(PGC-1α)在转录因子肝细胞核因子4α(HNF4α)介导下对人PEPCK基因启动子活性的影响【方法】从人全血基因组DNA中克隆PEPCK启动子基因片段，并重组进荧光素酶报告载体PGL3-basic，将pcDNA3.1-PGC-1αpcDNA3.0-HNF4α表达质粒与PGL3-hPCK-luc按不同组合共转染进人肝癌细胞株HepG2细胞或正常人肝细胞株LO2细胞,培养48 h后检测细胞裂解液中荧光素酶活性【结果】通过酶切鉴定及测序证明扩增...

Full description

Saved in:
Bibliographic Details
Published inZhongshan da xue xue bao. Zhongshan daxue xuebao yixue kexue ban = Journal of Sun Yat-sen University. Yi xue ke xue ban Vol. 32
Format Journal Article
LanguageChinese
Published Editorial Office of Journal of Sun Yat-sen University 01.01.2011
Subjects
基因表达调控
磷酸烯醇式丙酮酸羧激酶
糖异生
过氧化物酶体增殖物辅助激活因子1α
Online AccessGet full text
ISSN1672-3554

Cover

Abstract 【目的】构建人磷酸烯醇式丙酮酸羧激酶(PEPCK)基因启动子荧光素酶报告质粒PGL3-hPCK-luc，并探讨过氧化物酶体增殖物活化受体γ辅助激活因子1α(PGC-1α)在转录因子肝细胞核因子4α(HNF4α)介导下对人PEPCK基因启动子活性的影响【方法】从人全血基因组DNA中克隆PEPCK启动子基因片段，并重组进荧光素酶报告载体PGL3-basic，将pcDNA3.1-PGC-1αpcDNA3.0-HNF4α表达质粒与PGL3-hPCK-luc按不同组合共转染进人肝癌细胞株HepG2细胞或正常人肝细胞株LO2细胞,培养48 h后检测细胞裂解液中荧光素酶活性【结果】通过酶切鉴定及测序证明扩增的人PEPCK启动子片段成功插入载体质粒中转染后人PEPCK启动子基因荧光素酶活性较空白对照组增加60倍(P < 0.05)；共转染质粒进HepG2细胞或LO2细胞后，HNF4α均可以增强人PEPCK启动子基因荧光素酶活性，在HepG2细胞，荧光素酶活性是对照组的2.69倍(P < 0.05)，而在LO2细胞中，荧光素酶活性是对照组的3.64倍(P < 0.05)；PGC-1α可以明显增强HNF4α对人PEPCK启动子的激活作用，在HepG2细胞，荧光素酶活性是PGL3-HPCK-luc+HNF4α组的2.01倍(P < 0.05)，而在LO2细胞中,荧光素酶活性是PGL3-HPCK-luc+HNF4α组的2.82倍(P < 0.05)而与单一转染组相比，共转染pcDNA3.1-PGC-1α与pcDNA3.0-HNF4α显著增加了PEPCK的mRNA和蛋白表达水平(P < 0.05)【结论】人PEPCK启动子基因报告质粒成功构建，且转录因子HNF4α可以激活人PEPCK启动子活性，辅助激活因子PGC-1α可以进一步加强HNF4α的作用
AbstractList 【目的】构建人磷酸烯醇式丙酮酸羧激酶(PEPCK)基因启动子荧光素酶报告质粒PGL3-hPCK-luc，并探讨过氧化物酶体增殖物活化受体γ辅助激活因子1α(PGC-1α)在转录因子肝细胞核因子4α(HNF4α)介导下对人PEPCK基因启动子活性的影响【方法】从人全血基因组DNA中克隆PEPCK启动子基因片段，并重组进荧光素酶报告载体PGL3-basic，将pcDNA3.1-PGC-1αpcDNA3.0-HNF4α表达质粒与PGL3-hPCK-luc按不同组合共转染进人肝癌细胞株HepG2细胞或正常人肝细胞株LO2细胞,培养48 h后检测细胞裂解液中荧光素酶活性【结果】通过酶切鉴定及测序证明扩增的人PEPCK启动子片段成功插入载体质粒中转染后人PEPCK启动子基因荧光素酶活性较空白对照组增加60倍(P < 0.05)；共转染质粒进HepG2细胞或LO2细胞后，HNF4α均可以增强人PEPCK启动子基因荧光素酶活性，在HepG2细胞，荧光素酶活性是对照组的2.69倍(P < 0.05)，而在LO2细胞中，荧光素酶活性是对照组的3.64倍(P < 0.05)；PGC-1α可以明显增强HNF4α对人PEPCK启动子的激活作用，在HepG2细胞，荧光素酶活性是PGL3-HPCK-luc+HNF4α组的2.01倍(P < 0.05)，而在LO2细胞中,荧光素酶活性是PGL3-HPCK-luc+HNF4α组的2.82倍(P < 0.05)而与单一转染组相比，共转染pcDNA3.1-PGC-1α与pcDNA3.0-HNF4α显著增加了PEPCK的mRNA和蛋白表达水平(P < 0.05)【结论】人PEPCK启动子基因报告质粒成功构建，且转录因子HNF4α可以激活人PEPCK启动子活性，辅助激活因子PGC-1α可以进一步加强HNF4α的作用
BookMark eNotjLFKw1AUQN9QwVr7HU6BvPvefS93lFBrsWAGncO9TSKR2kji4ubi4u7koKvgInSKVP_ArzCln6GoZzlwhrOjeotqkfdUXzsPgUG022rYNKWEoUNnwFJf7SXjONCfr5v32-7uef1xs16-fbVtMkrio-6x7R6eNquXbnW_q7YKnjf58N8DdXowOokPg-nxeBLvT4OZBk-BaHGmQCq0Jut1SALkEaMZETvrPBXsAVl7bwgdiHhnkbEgyBDNDwM1-ftmFZ-nl3V5wfV1WnGZ_oaqPku5vipn8zzF0HMkOTiNYllIosygyQQYjBgC8w2HPVIg
ContentType Journal Article
DBID DOA
DatabaseName DOAJ Directory of Open Access Journals
DatabaseTitleList
Database_xml – sequence: 1
  dbid: DOA
  name: DOAJ Directory of Open Access Journals
  url: https://www.doaj.org/
  sourceTypes: Open Website
DeliveryMethod fulltext_linktorsrc
Discipline Medicine
ExternalDocumentID oai_doaj_org_article_507a8be2615b4ab9b8d353db2a23b392
GroupedDBID ALMA_UNASSIGNED_HOLDINGS
GROUPED_DOAJ
ID FETCH-LOGICAL-c1279-b1b63f59f11947109b297558c99a64679fa725a17739562bb7645a5f92d553333
IEDL.DBID DOA
ISSN 1672-3554
IngestDate Fri Oct 03 12:52:28 EDT 2025
IsOpenAccess true
IsPeerReviewed true
IsScholarly true
Language Chinese
LinkModel DirectLink
MergedId FETCHMERGED-LOGICAL-c1279-b1b63f59f11947109b297558c99a64679fa725a17739562bb7645a5f92d553333
OpenAccessLink https://doaj.org/article/507a8be2615b4ab9b8d353db2a23b392
ParticipantIDs doaj_primary_oai_doaj_org_article_507a8be2615b4ab9b8d353db2a23b392
PublicationCentury 2000
PublicationDate 2011-01-01
PublicationDateYYYYMMDD 2011-01-01
PublicationDate_xml – month: 01
  year: 2011
  text: 2011-01-01
  day: 01
PublicationDecade 2010
PublicationTitle Zhongshan da xue xue bao. Zhongshan daxue xuebao yixue kexue ban = Journal of Sun Yat-sen University. Yi xue ke xue ban
PublicationYear 2011
Publisher Editorial Office of Journal of Sun Yat-sen University
Publisher_xml – name: Editorial Office of Journal of Sun Yat-sen University
SSID ssib006563249
ssib001051178
ssib002039699
ssj0002511113
ssib006703418
ssib001103819
ssib002262833
ssib051369397
ssib001186722
ssib043242460
ssib000970529
Score 1.8480874
Snippet 【目的】构建人磷酸烯醇式丙酮酸羧激酶(PEPCK)基因启动子荧光素酶报告质粒PGL3-hPCK-luc，并探讨过氧化物酶体增殖物活化受体γ辅助激活因子1α(PGC-1α)在转录因子肝细胞核因...
SourceID doaj
SourceType Open Website
SubjectTerms 基因表达调控
磷酸烯醇式丙酮酸羧激酶
糖异生
过氧化物酶体增殖物辅助激活因子1α
Title PGC-1α辅助激活人PEPCK基因转录
URI https://doaj.org/article/507a8be2615b4ab9b8d353db2a23b392
Volume 32
hasFullText 1
inHoldings 1
isFullTextHit
isPrint
link http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwrZ07SwNBEMcXsRAb8YlvUghWi9nn3ZYaEoMSSWEgXdi5B1ZRJDZWNjb2VhbaCjaCVST6DfwUXsjHcCd3kLWyEa66Yo_b2Zv5zc3ufwjZASONlommPBWKuojPqY0TTl16GxqNClAWDwo3TnS9JY_aqu21-sI9Ybk8cD5xe45XbAiJA30F0oKBMBZKxMAtF-CCO3rfcmi8ZCoHhwBLWF7i48DCE65iqAvuC5mhrpsnZMfLwmgf1LnmoQdGjoKEn7ho9-HIiZdHmTsuJ4mHYkIbUdTrMEYg2LNx72bmnksx6P9qGjCObrV5MldgaWk_n44FMnV9tkhmGkXhfYnsNg8rlH29jj5us7vn4efN8O39u99vVpuV4-yxnz08jQYv2eB-mbRq1dNKnRY9FmjEeGAoMNAiVSZlzEjclwl41FaFkTFWOydqUhtwZVmABT3NAQItlVWp4bFypCjECpnunneTVVJyzkJGAi0fOUywALEUMQ4ZJUIHoNfIAb5g5yKX0eigsPX4hjN3pzB35y9zr__HIBtkNv81jNcmme5dXiVbji16sD1eRj_pzsHD
linkProvider Directory of Open Access Journals
openUrl ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=PGC-1%CE%B1%E8%BE%85%E5%8A%A9%E6%BF%80%E6%B4%BB%E4%BA%BAPEPCK%E5%9F%BA%E5%9B%A0%E8%BD%AC%E5%BD%95&rft.jtitle=Zhongshan+da+xue+xue+bao.+Zhongshan+daxue+xuebao+yixue+kexue+ban+%3D+Journal+of+Sun+Yat-sen+University.+Yi+xue+ke+xue+ban&rft.date=2011-01-01&rft.pub=Editorial+Office+of+Journal+of+Sun+Yat-sen+University&rft.issn=1672-3554&rft.volume=32&rft.externalDBID=DOA&rft.externalDocID=oai_doaj_org_article_507a8be2615b4ab9b8d353db2a23b392
thumbnail_l http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=1672-3554&client=summon
thumbnail_m http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=1672-3554&client=summon
thumbnail_s http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=1672-3554&client=summon