Short Hairpin RNA-mediated MDR1 Gene Silencing Increases Apoptosis of Human Ovarian Cancer Cell Line A2780Taxol.pdfVIP

】38 ChinJCancerRes24(2)：138-142,2012 www．springerlink．cam OriginalArticle ShortHairpinRNA—-mediatedMDR1G ApoptosisofHumanOvarianCancerC HuiXu Fan—zhenHong SuLi PingZhang LinZhu DepartmentD，ObstetricsandGynecology,theSecondHospitalof5handongUniversity,Jinan250033,China DOI：10．1007／s11670—012—0138—3 ~ChineseAnti—-CancerAssociationandSpringer--VerlagBerlinHeidelberg2012 ABSTRACT Objective：RecurrentovariancancerisoftenresistanttodrugssuchaspaclitaxeI．ShorthairpinRNA (shRNA) targetingMDR1，agene involved intheprocessofdrugresistance，maybeapromisingstrategytoovercomedrug resistance． M ethods：ConstructionandidentificationofeukaryoticexpressionplasmidofshRNAtargetingonMDR1gene．The plasmidwastransientlytransfectedintohumanovariancancercelIIiReA2780／Taxo1．Apoptosiswasdeterminedby flow cytometryusingannexinV．FITC／PIdoubleIabeling．ExpressionofMDR工mRNAwasdetectedbyquantitative polymerasechainreaction(qPCR)andp-glycoproteinexpressionwasdetectedusingWesternblot． Results：TheIC50ofpaclitaxelinMD尺IshRNA—transfectedgroupwassignificantlyreduced(1．986-+0．153)pmol／ml ascomparedwiththatinnegativecontroI(5．246+0．107)IJ．mol／mlandemptyvector—transfectedgroup(5．212+0．075) moI／mI(PO．05)．ThepercentoftherelativereversesensitivitytopaclitaxelonA2780／Taxolcellswas67．1％，andthe apoptoticratewassignificantlyincreased (【6．977+0．333)％]comparedwithcontrol【(1．637+0．111)％]andempty vector—transfectedgroup[(1．663-+0．114)％】(P0．0s)．ExpressionsofMDRImRNAandp-glycoproteinweresignificantly reducedcomparedwithcontrol(PO．os)． Conclusion：ThepresentstudydemonstratedthattheeukaryoticexpressionplasmidofshRNA targetingonMDR1 inhibitedtheexpressionofMDR1effectively,thusenhancethesensitivityofA2780／TalxolcellstopacIitaxe1． Keywords：Ovariancancer,ShorthairpinRNA，Paclitaxe