hsvtk基因治疗靶向载体的构建及特异性表达分析

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1、万方数据《癌症》ChineseJournalofCancer，2006，25(2)：179-184179HSV．tk基因治疗靶向载体的构建及特异性表达分析刘晓平1，李宝金1，张超2ConstructionandIdentificationofRecombinantVectorsCarryingHerpesSimplexVirusThymidineKinaseGenesExpressedinVascularEndothelialCellsLIUXiao—Pin91，LIBao—Jinl，ZHANGCha021．北京大学深圳

2、医院肝胆胰外科。广东深圳5180362．深圳北京大学香港科技大学医学中心。广东深圳518036，．DepartmentofHepatobilliaryandPancreaticSurgery，ShenzhenHospital，PekingUniversity，Shenzhen，Guangdong，518036，P．R．China2．ShenzhenPKU—HKUsTMedic击Ceriter，Shenzhen，Guangdong，518036，P．R．Chjina通讯作者：刘晓平Correspondenceto：LIUX

3、iao—PingTel：86—755—83923333—6026Fax：86—755—83061340E—mail：ninliu@163．com基金项目：国家自然科学基金资助项目(No．30371386)；广东省自然科学基金资助项目(No．31010)Grants：NationalNaturalScienceFoundationofChina(No．30371386)；NaturalScienceFoundationofGuangdongProvince(No．31010)收稿日期：2005—02．25修回日期：2005

4、—10—12基础研究·[ABSTRACT]BACKGROUND＆OBJECTIVE：Blockingtumorangiogenesisisapromisingtumoricidalapproach．Herpessimplexvirus—thymidinekinase(HSV—tk)suicidegenecouldeffectivelydamageneoplasticvascularendotheliaIcells．However．thekillingeffectofcurrentlyusedHSV—tksystemisn

5、on—specificduetoemployingcytomegalovirus(CMV)asitspromoter．Kinasedomaininsertcontainingreceptor(KDR)，akindofvascularendothelialgrowthfactorreceptor，ishighlyexpressedontumorvascularendothelialcells．butlowlyexpressedonnormaItissues．ThisstudywastOconstructarecombina

6、ntadenoviruscontainingKDRpromoter—mediatedHSV—tksuicidegene．andanalyzeitsspecificexpressiononvascularendothelialcells．MEAHODS：KDR—tkfragmentwassubclonedintocorrespondingrestrictionsitesofvectorwithdirectionalcloningmethodusingpAdeasysystemtoconstructrecombinantad

7、enoviraIplasmidcontainingKDRpromoter—controlledHSV—tkgene(AdKDR—tk)．Afterpackagedandamplifiedin293cells．theviruswasusedtOinfectKDR—expressedhumanumbiticalvenousendothelialcells(HUVECs)andKDR—unexpressedlivercancercelIlineHepG2．Afteradministrationofganciclovir(GCV

8、)，proliferationofgene—transfectedHUVECsandHepG2cellswasevaluatedbvM丌assay．RESULTS：Accordingtotheresultsofrestrictiondigestion，reversetranscription—polymerasech