RegIα cDNA过表达质粒的构建及对胃癌细胞增殖凋亡的影响

Q78; 目的:构建再生蛋白Iα(regeneration gene Iα,RegIα)的cDNA过表达质粒表达载体,在体外评价其对胃癌细胞株增殖及其凋亡的影响.方法:根据RegIα cDNA的全序列设计引物,用RT-PCR的方法从胃黏膜组织的总RNA中获取RegIα的cDNA编码区域的全序列,经测序其与目的基因序列一致.将获取的RegIα序列经TA克隆和亚克隆至pIRES2-EGFP真核表达质粒载体中;将构建的RegIα过表达质粒载体pIRES2-RegIα-EGFP转染胃癌细胞株MKN28,并用空载体pIRES2-EGFP作为对照,利用G418筛选稳定表达pIRES2-RegIα-EGFP...

Full description

Saved in:
Bibliographic Details
Published in浙江大学学报（医学版） Vol. 39; no. 5; pp. 499 - 505
Main Authors 周群燕, 陆晓凤, 王良静, 姒健敏
Format Journal Article
LanguageChinese
Published 浙江大学邵逸夫临床医学研究所胃肠病实验室,浙江,杭州,310016%浙江大学医学院附属第二医院消化内科,浙江,杭州,310009 2010
Subjects
细胞凋亡
逆转录聚合酶链反应
再生/遗传学
转染
遗传载体
细胞系,肿瘤
胃肿瘤
质粒
细胞增殖
Online AccessGet full text
ISSN1008-9292
DOI10.3785/j.issn.1008-9292.2010.05.010

Cover

Abstract Q78; 目的:构建再生蛋白Iα(regeneration gene Iα,RegIα)的cDNA过表达质粒表达载体,在体外评价其对胃癌细胞株增殖及其凋亡的影响.方法:根据RegIα cDNA的全序列设计引物,用RT-PCR的方法从胃黏膜组织的总RNA中获取RegIα的cDNA编码区域的全序列,经测序其与目的基因序列一致.将获取的RegIα序列经TA克隆和亚克隆至pIRES2-EGFP真核表达质粒载体中;将构建的RegIα过表达质粒载体pIRES2-RegIα-EGFP转染胃癌细胞株MKN28,并用空载体pIRES2-EGFP作为对照,利用G418筛选稳定表达pIRES2-RegIα-EGFP的MKN28细胞株.通过RT-PCR及Western blot检测转染后细胞RegIα在mRNA及蛋白质水平的表达;MTT法和流式细胞仪分别检测RegIα过表达后MKN28细胞增殖及拮抗H2O2诱导凋亡的变化.结果:成功构建RegIα cDNA的全序列表达质粒.RT-PCR及Western blot检测显示:转染pIRES2-EGFP质粒的MKN28细胞RegIα mRNA及蛋白表达水平明显升高(P<0.05);MTT试验显示:转染后细胞增殖能力明显升高(P<0.05),而且拮抗H2O2凋亡的能力增强(P<0.05).结论:RegIα的过表达能明显增强胃癌细胞MKN28的增殖和拮抗H2O2诱导凋亡的能力.
AbstractList Q78; 目的:构建再生蛋白Iα(regeneration gene Iα,RegIα)的cDNA过表达质粒表达载体,在体外评价其对胃癌细胞株增殖及其凋亡的影响.方法:根据RegIα cDNA的全序列设计引物,用RT-PCR的方法从胃黏膜组织的总RNA中获取RegIα的cDNA编码区域的全序列,经测序其与目的基因序列一致.将获取的RegIα序列经TA克隆和亚克隆至pIRES2-EGFP真核表达质粒载体中;将构建的RegIα过表达质粒载体pIRES2-RegIα-EGFP转染胃癌细胞株MKN28,并用空载体pIRES2-EGFP作为对照,利用G418筛选稳定表达pIRES2-RegIα-EGFP的MKN28细胞株.通过RT-PCR及Western blot检测转染后细胞RegIα在mRNA及蛋白质水平的表达;MTT法和流式细胞仪分别检测RegIα过表达后MKN28细胞增殖及拮抗H2O2诱导凋亡的变化.结果:成功构建RegIα cDNA的全序列表达质粒.RT-PCR及Western blot检测显示:转染pIRES2-EGFP质粒的MKN28细胞RegIα mRNA及蛋白表达水平明显升高(P<0.05);MTT试验显示:转染后细胞增殖能力明显升高(P<0.05),而且拮抗H2O2凋亡的能力增强(P<0.05).结论:RegIα的过表达能明显增强胃癌细胞MKN28的增殖和拮抗H2O2诱导凋亡的能力.
Author 周群燕
陆晓凤
王良静
姒健敏
AuthorAffiliation 浙江大学邵逸夫临床医学研究所胃肠病实验室,浙江,杭州,310016%浙江大学医学院附属第二医院消化内科,浙江,杭州,310009
AuthorAffiliation_xml – name: 浙江大学邵逸夫临床医学研究所胃肠病实验室,浙江,杭州,310016%浙江大学医学院附属第二医院消化内科,浙江,杭州,310009
Author_FL LU Xiao-feng
WANG Liang-jing
ZHOU Qun-yan
SI Jian-min
Author_FL_xml – sequence: 1
  fullname: ZHOU Qun-yan
– sequence: 2
  fullname: LU Xiao-feng
– sequence: 3
  fullname: WANG Liang-jing
– sequence: 4
  fullname: SI Jian-min
Author_xml – sequence: 1
  fullname: 周群燕
– sequence: 2
  fullname: 陆晓凤
– sequence: 3
  fullname: 王良静
– sequence: 4
  fullname: 姒健敏
BookMark eNrjYmDJy89LZWBQMTTQMza3MNXP0sssLs7TMzQwsNC1NLI00jMyAEoZmOoBKRYGTrg4BwNvcXFmkoGpobE5EBpwMngFpaZ7ntuokOzi5_hif_uLhSte7Nv3YsuK55smPZ_V8mxey9Pdu572dz1dv_NFc_PzmT3Pd7e9aJ73dNG8Z-umPW3vfrJrIVDZ070bn07u5WFgTUvMKU7lhdLcDCFuriHOHro-_u6ezo4-uslmxga6qcamqaYmJinJJmlGBmmpKWYmaRbGloaphpaJZmkGKYYG5iamaWZANWaGFolJKUZphqaphqbGFibmRmmJZompxtwM2hBjyxPz0hLz0uOz8kuL8oAWxldlpVRUJMVXViSBvG9gCiSMAWsWa0g
ClassificationCodes Q78
ContentType Journal Article
Copyright Copyright © Wanfang Data Co. Ltd. All Rights Reserved.
Copyright_xml – notice: Copyright © Wanfang Data Co. Ltd. All Rights Reserved.
DBID 2B.
4A8
92I
93N
PSX
TCJ
DOI 10.3785/j.issn.1008-9292.2010.05.010
DatabaseName Wanfang Data Journals - Hong Kong
WANFANG Data Centre
Wanfang Data Journals
万方数据期刊 - 香港版
China Online Journals (COJ)
China Online Journals (COJ)
DatabaseTitleList
DeliveryMethod fulltext_linktorsrc
Discipline Medicine
DocumentTitle_FL Construction of RegIα over-expression vector and its effect on proliferation and apoptosis of gastric cancer cells
EndPage 505
ExternalDocumentID zjdxxb_yxb201005010
GrantInformation_xml – fundername: 浙江省自然科学基金; 浙江省教育厅科研项目
  funderid: (Y206280); (Y20070071)
GroupedDBID 2B.
4A8
92I
93N
ALMA_UNASSIGNED_HOLDINGS
PSX
RPM
TCJ
ID FETCH-LOGICAL-c630-e35e544dc4f20fed64f8391e19a6f0d10745f65e5618abd2f15e1538472fa6ae3
ISSN 1008-9292
IngestDate Thu May 29 04:07:20 EDT 2025
IsPeerReviewed false
IsScholarly true
Issue 5
Keywords 细胞凋亡
逆转录聚合酶链反应
再生/遗传学
转染
遗传载体
细胞系,肿瘤
胃肿瘤
质粒
细胞增殖
Language Chinese
LinkModel OpenURL
MergedId FETCHMERGED-LOGICAL-c630-e35e544dc4f20fed64f8391e19a6f0d10745f65e5618abd2f15e1538472fa6ae3
PageCount 7
ParticipantIDs wanfang_journals_zjdxxb_yxb201005010
PublicationCentury 2000
PublicationDate 2010
PublicationDateYYYYMMDD 2010-01-01
PublicationDate_xml – year: 2010
  text: 2010
PublicationDecade 2010
PublicationTitle 浙江大学学报（医学版）
PublicationTitle_FL JOURNAL OF ZHEJIANG UNIVERSITY(MEDICAL SCIENCES)
PublicationYear 2010
Publisher 浙江大学邵逸夫临床医学研究所胃肠病实验室,浙江,杭州,310016%浙江大学医学院附属第二医院消化内科,浙江,杭州,310009
Publisher_xml – name: 浙江大学邵逸夫临床医学研究所胃肠病实验室,浙江,杭州,310016%浙江大学医学院附属第二医院消化内科,浙江,杭州,310009
SSID ssib051373730
ssib002263662
ssib009993744
ssib056826211
ssib058720545
ssib006576290
ssib006704889
ssj0002507821
ssib006562844
ssib002039758
ssib008143987
ssib008679799
ssib001051301
ssib001104240
Score 1.8339158
Snippet Q78; 目的:构建再生蛋白Iα(regeneration gene Iα,RegIα)的cDNA过表达质粒表达载体,在体外评价其对胃癌细胞株增殖及其凋亡的影响.方法:根据RegIα cDNA的全序列设计引物,...
SourceID wanfang
SourceType Aggregation Database
StartPage 499
Title RegIα cDNA过表达质粒的构建及对胃癌细胞增殖凋亡的影响
URI https://d.wanfangdata.com.cn/periodical/zjdxxb-yxb201005010
Volume 39
hasFullText 1
inHoldings 1
isFullTextHit
isPrint
journalDatabaseRights – providerCode: PRVHPJ
  databaseName: ROAD: Directory of Open Access Scholarly Resources
  issn: 1008-9292
  databaseCode: M~E
  dateStart: 19990101
  customDbUrl:
  isFulltext: true
  dateEnd: 99991231
  titleUrlDefault: https://road.issn.org
  omitProxy: true
  ssIdentifier: ssib056826211
  providerName: ISSN International Centre
link http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnR3LahRBcAgGohfxiW9ySJ9k47y6p_s4MztLDCQHiZBbmGfEwwqaQMzBS4IBHwgKgh4C8ZSTEvFigvEP_Ao3u59hVc1kpmOCJLLDUumtrmenp6q2ptcwxoSXpamTWq24UFkLdj_VipPEaTmJKVWauE4WY71jalpM3HcnZ_ns0PBprWtpcSEZT5ePfK7kf7wKY-BXfEr2BJ6ticIAwOBfeAcPw_uxfHwvn7_LwogF1u20Pe2zSLKgwyC9B8C3mC9pJMILAZdGPBbYTNkIKJ9Jl0WCqYgAzoKABT4CEugQ4HdYoHC6dPDCWYrJkOgETIrqI6CAyDYBgvkRU4LoeEwGLHKRLIjUMAVebRAcAQWU23qUjBQCjowQsJjqEHGQ3yOgzXyhAQJF9TmLQNSQSUl8QxSvwQExFH1U4tTlD-Jv7RsmQiaIC2LyBkWRzsQJABC3VMx3GxSYE5GmEhmB0XBSm6kDjEB8NDxMtkheIIeW1isvVfct_pcczwogmI1oCJgskPs4pdHJ62hrv3LnYbv4JlOktK9YUFqTSDUul8gC0NB5oCen6RF5GlT1aTXQCPC1wyPELgfBHMAUhfFwrh3itz5Y1OHH9fffwpNfYM1VDio1tUgwUCfEq1x5pYTV9HqWQH3r5VKtV0EKeuQr68TqmEq7y2PPD-QFB8IAR2nbHdfu6W61VMrwkNMpAYciD8eTnCIPZDBeM6h6R_FoYLOJuOo-2OWH2dJSMvd0KUE8k9PDosM2lhaxC_lZpFVuOESb-kmN2LOgRe4mJBZagwNkVY7QTgKFtA0CUVf7G6Ig7Qty4eGtvMlsJORRSsvc8FxOvQFAYV7T0APhPHjV9LiQtrCb0IFLz4bkkNflb8i8IDehcti-rUaMscqQd_5lRnq6slvE3XktEZg5Z5ytMvhRv9yOzxtDyw8uGCNTVY_SRWMSd-VfW6O4Iw9-rg02Nge7u4Nvm_2vb_sfV_fWV3s72703L3pfvg9WVvofXvV3ng9W1nuf1vc-v--tvfy9vQFovR9bvXevLxkznWgmnGhVv1jTSoVjtnKH59x1s9QtbLPIM-EWkH9auaViUZgZ9r7zQgCOsGScZHZh8RwjTtezi1jEuXPZONV91M2vGKNxJrBjJHccJ3FTCCu92EwzlWEXR24V9lVjrLLCXHVDejJ3xGq6djy068aZsisKS8s3jFMLjxfzm5BsLSS3aBn-AbgzDz4
linkProvider ISSN International Centre
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=RegI%CE%B1+cDNA%E8%BF%87%E8%A1%A8%E8%BE%BE%E8%B4%A8%E7%B2%92%E7%9A%84%E6%9E%84%E5%BB%BA%E5%8F%8A%E5%AF%B9%E8%83%83%E7%99%8C%E7%BB%86%E8%83%9E%E5%A2%9E%E6%AE%96%E5%87%8B%E4%BA%A1%E7%9A%84%E5%BD%B1%E5%93%8D&rft.jtitle=%E6%B5%99%E6%B1%9F%E5%A4%A7%E5%AD%A6%E5%AD%A6%E6%8A%A5%EF%BC%88%E5%8C%BB%E5%AD%A6%E7%89%88%EF%BC%89&rft.au=%E5%91%A8%E7%BE%A4%E7%87%95&rft.au=%E9%99%86%E6%99%93%E5%87%A4&rft.au=%E7%8E%8B%E8%89%AF%E9%9D%99&rft.au=%E5%A7%92%E5%81%A5%E6%95%8F&rft.date=2010&rft.pub=%E6%B5%99%E6%B1%9F%E5%A4%A7%E5%AD%A6%E9%82%B5%E9%80%B8%E5%A4%AB%E4%B8%B4%E5%BA%8A%E5%8C%BB%E5%AD%A6%E7%A0%94%E7%A9%B6%E6%89%80%E8%83%83%E8%82%A0%E7%97%85%E5%AE%9E%E9%AA%8C%E5%AE%A4%2C%E6%B5%99%E6%B1%9F%2C%E6%9D%AD%E5%B7%9E%2C310016%25%E6%B5%99%E6%B1%9F%E5%A4%A7%E5%AD%A6%E5%8C%BB%E5%AD%A6%E9%99%A2%E9%99%84%E5%B1%9E%E7%AC%AC%E4%BA%8C%E5%8C%BB%E9%99%A2%E6%B6%88%E5%8C%96%E5%86%85%E7%A7%91%2C%E6%B5%99%E6%B1%9F%2C%E6%9D%AD%E5%B7%9E%2C310009&rft.issn=1008-9292&rft.volume=39&rft.issue=5&rft.spage=499&rft.epage=505&rft_id=info:doi/10.3785%2Fj.issn.1008-9292.2010.05.010&rft.externalDocID=zjdxxb_yxb201005010
thumbnail_s http://utb.summon.serialssolutions.com/2.0.0/image/custom?url=http%3A%2F%2Fwww.wanfangdata.com.cn%2Fimages%2FPeriodicalImages%2Fzjdxxb-yxb%2Fzjdxxb-yxb.jpg