MicroRNA-10b对宫颈癌细胞侵袭迁移能力的影响及机制研究

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1、MicroRNA-lOb对宫颈癌细胞侵袭迁移能力的影响及机制研究陶贝贝周文娟张婷郑州市妇幼保健院妇产科郑州大学第三附属医院妇产科摘要：目的:探讨微小RNA-lOb(microRNA-10b)对宫颈癌细胞侵袭迁移能力的影响及机制。方法:实时定量聚合酶链反应(real-timePCR)检测microRNA-10b在不同宫颈癌细胞及正常宫颈上皮细胞中的表达情况;脂质体转染microRNA-10b模拟物(microRNA-10bmimics,即mimics组)及microRNA无关序列(microRNA-10bNC,即NC组)至Caski细胞，转染micr

2、oRNA-10b抑制物(microRNA-10binhibitor,即inhibitor组)及microRNAinhibitor无关序列(microRNA-10bNCinhibitor,即NCinhibitor组)至HeLa细胞,转染12h后，real-timePCR法检测转染效率。采用Transwel1试验和划痕试验分别检测microRNA-10b对Caski和HeLa细胞侵袭迁移能力的影响;real-timePCR及蛋口质印迹(Westernblotting)检测各组细胞中胰岛素样生长因子1受体(1GF-1R)mRNA和蛋白表达水平；荧光素酶报

3、告基因试验检测microRNA-10b对IGF-1R基因表达的调控机制。结果:MicroRNA-lOb在宫颈癌细胞中表达水平低于正常宫颈上皮细胞(P<0.05)oMicroRNA-lOb表达增加能够降低Caski细胞的侵袭迁移能力(P<0.05),抑制IGF-1R的mRNA和蛋白表达(均P<0.05);而microRNA-10b表达下降能够增强HeLa细胞的侵袭迁移能力(P<0.05),增加1GF-1R的mRNA和蛋白表达(P均<0.05)。在HEK-293T细胞中，microRNA-10b表达增加能够减弱含有IGF-1R基因3,非翻译区(3,-U

4、TR)的报告质粒的荧光素酶活性(P<0.05)<>结论:在宫颈癌细胞中,microRNA-10b通过下调TGF-1R基因表达，抑制细胞侵袭迁移能力。关键词：微RNAs;宫颈肿瘤;肿瘤侵润;细胞运动;收稿日期：2017-07-18TheEffectsofMicroRNA-10bontheInvasionandMigrationintheCervicalCarcinomaCellsandItsPossibleMechanismTAOBei-beiZHOUWen-juanZHANGTingDepartmentofObstetricsandGynecolo

5、gy,Women&InfantsHospitalZhengzhou;DepartmentofObstetricsandGynecology,TheTh让dAff订iatedHospitalofZhengzhouUniversity;Abstract：Objective:ToinvestigatetheroleofmicroRNA-10binthecervicalcarcinomaandexploreitspossiblemechanism.Methods:TheexpressionlevelsofmicroRNA-lObincervicalcarc

6、inomacelllinesandthenormalcervicalepithelialcellsweredetectedbythereal-timepolymerasechainreaction(real-timePCR).ThemicroRNA-10bmimics/NCwastransfectedintotheCaskicellsandthemicroRNA-10binhibitor/NCinhibitorwastransfectedintotheHeLacellsbytheliposome.TheTranswellassayandwound-

7、healingassaywereusedtodeterminetheeffectsofmicroRNA-10bontheinvasionandmigrationabilityintheCaskiandHeLacells.ThemRNAandproteinlevelsofinsulin-likegrowthfactor1receptor(IGF~1R)wereanalyzedbythereal-timePCRandtheWesternblottingassay,respectively.Luciferasereporterassaywasapplie

8、dtodetecttheregulatorymechanismofmicroRNA-10bintheIGF-1Rgene.