低相对分子质量肝素联合多柔比星对肝癌细胞迁移能力的影响-论文.pdf

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1、·工D48·doi10．3969／j．issn．1673—4254．2014．07．27JSouthMedUniv,2014，34(7)：1048—1052基础研究低相对分子质量肝素联合多柔比星对肝癌细胞迁移能力的影响杨克红，马琳艳，程秀，陈超，张梦晓，刘浩，蒋志文蚌埠医学院药学系／／安徽省生化药物工程技术研究中心，安徽蚌埠233030摘要：目的探讨低相对分子质量肝素(LMwH)联合多柔比星(DxRB)对肝癌细胞迁移能力的影响及其作用机制。方法实验分为阴性对照组、LMWH组、DXRB组、LMWH与DXRB联合应用组。用MTT法检测药物对HepG2细胞增殖的影响；用细胞划痕实验和T

2、ranswell／J~室法~JHepG2细胞的迁移能力；Western-blot和实时RT-PCR法检测基质金属蛋白酶．9(MM．P．9)、基质金属蛋白酶．2(MMP．2)的表达；ELISA法检测肿瘤细胞培养液中乙酰肝素酶的表达。结果MTT结果显示，LMWH和DXRB对肝癌HepG2细胞均有显著的增殖抑制作用。LMWH与DXRB联合用药时，对肝癌HepG2细胞的迁移抑制作用明显高于单用DXRB组。LMWH和DXRB均可下调MMP．9、MMP．2和乙酰肝素酶的表达，且合用时对乙酰肝素酶表达的抑制作用更明显。结论LMWH具有增强DXRB抑制肝癌HepG2细胞迁移的作用，其机制可能与下

3、调MMP．9、MMP．2和乙酰肝素酶的表达有关。关键词：肝癌；低相对分子质量肝素；多柔比星；迁移；基质金属蛋白酶一9；基质金属蛋白酶一2；乙酰肝素酶Effectoflow—molecular-weightheparincombinedwithdoxorubicinonhepatocellularcancercellmigrationinvitroYANGKehong,MALinyanCHENGXiu

4、CHENChao,ZHANGMengxiao,LIUHao,JIANGZhiwenDepartmentofPharmacy,BengbuMedicalCollege,AnhuiEng

5、ineeringTechnologyResearchCenterofBiochemicalPharmaceuticals,Bengbu233030_CAbstract：ObjectiveToinvestigatetheanti—cancereffectof1OW—molecular-weightheparin(LMWH1combinedwithdoxorubicinandexplorethemechanism．MethodsHepatocellularcancerHepG2cellsexposedtoLMWH，doxorubicin,orbothwereevaluatedforc

6、ellviabilitywithmTassayandforchangesintheirmigrationabilityusingwoundhealingassayandTranswellmigrationassay．nchangesincellularexpressionsofmatrixmetalloproteinase一9(MMP一91andMMP一2mRNAandproteinswereanalyzedwithquantitativereal—timePCR(qRT—PCR)andWstemblotting,andELISAwasusedtodetermineheparan

7、ase(HPA)concentrationinthecellculturemedium．ResultsHepG2cellsexhibitedsuppressedproliferationinresponsetoLMWHanddoxorubicintreatments．ThecombinedtreatmentcausedasignificantlyhigherinhibitionrateofcellmigrationthanLMwHanddoxorubicinalone．LMWHenhanceddoxorubicin—induceddown．regulationofMMP_9，MM

8、P一2andHPAinthecells．ConclusionLMWHcanenhancetheinhibitoryeffectofdoxorubicinonthemigrationofHepG2cells,themechanismofwhichmayinvolvethedown．reg-ulationofMMP-9，MMP一2andHPAexpressions．Keywords：hepatocellularcancer；low—molecular-weighthepafin；do