hsamir27a和hsamir451在恶性肿瘤耐药细胞中的表达及与耐药关系的研究-expression of h samir 27a and h samir 451 in malignant tumor drug-resistant cells and their relationship with drug resistance.docx

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2018-05-29 发布于上海
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hsamir27a和hsamir451在恶性肿瘤耐药细胞中的表达及与耐药关系的研究-expression of h samir 27a and h samir 451 in malignant tumor drug-resistant cells and their relationship with drug resistance.docx

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hsamir27a和hsamir451在恶性肿瘤耐药细胞中的表达及与耐药关系的研究-expression of h samir 27a and h samir 451 in malignant tumor drug-resistant cells and their relationship with drug resistance

ExpressionofmiR-27aandmiR-451andrelationshipwithdrugresistanceinmalignanttumorAbstractPartⅠTheexpressionofmiR-27aandmiR-451indrugresistantcancercellswithhighexpressionofP-glycoproteinObjectiveToinvestigatetheexpressionofmiR-27aandmiR-451inovariancancerandbreastcancercells.MethodsA2780/Taxolcellswereestablishedusingstepwiseselection.TheexpressionofP-gpproteinlevelsofovariancancerandbreastcancercellsweremeasuredbywesternblot.Stem-loopreal-timePCRwasusedtodetectexpressionofmiR-27aandmiR-451inovariancancerandinbreastcancercells.ResultsA2780/Taxolcelllinewasestablishedsuccessfully.TheexpressionlevelsofP-gpwerehighinA2780/TaxolandMCF-7/ADMcellsanditdidnotdetectedinA2780andMCF-7cells.TheexpressionofmiR-27awasanaverageof(2.2±0.30)-foldhigherinA2780/TaxolcellsthaninA2780cells,withasignificantdifferencebetweenthetwogroups(P0.05).TheexpressionofmiR-451wasanaverageof(16±0.21)-foldlesserinMCF-7/ADMcellsthaninMCF-7cells,withasignificantdifferencebetweenthetwogroups(P0.05).ConclusionTheexpressionsofmiR-27aandmiR-451werederegulationinA2780/TaxolandMCF-7/ADMcellsrespectively.keywordsmiRNA；A2780/Taxol；MCF-7/ADM；P-gp;drugresistance.PartⅡStudytherelationshipbetweenmiR-27aanddrugresistantinpaclitaxel-resistantovariancancercellsObjectiveToinvestigatetherelationshipbetweenmiR-27aanddrugresistantinpaclitaxel-resistantovariancancercells.MethodsTheA2780andA2780/TaxolcellsweretransfectedwiththemimicsorinhibitorsofmiR-27aornegativecontrolRNA(NC)byLipofectamine2000.TheexpressionsofMDR1mRNA,P-gpandHIPK2proteinlevelsweremeasuredbyreal-timePCRandwesternblotrespectively.MTTwasusedtoanalyzedrugsensitivity.ApoptosisanalysisandthefluorescenceofremainingRh-123weremeasuredbyfluorescenceactivatedcellsorter(FACS).Results(1)A2780/TaxolcellstransfectionwithinhibitorsofmiR-27ashowedthatthelevelsofMDR1mRNAwasdecreasedby(39±0.14)%,P-gpproteinlevel((26±5.3）%)decreasedthantheNCgroup(（43±6.7）%),HIPK2proteinlevel(（30±5.9）%)increasedthantheNCgroup(（19±3.8）%),theIC50(0.53μM)waslessthantheNCgroup(6.8μM),apoptosisrate