资源描述：
《特异性ilk-sirna对膀胱癌ej细胞侵袭和转移的影响》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、特异性ILK-siRNA对膀胱癌EJ细胞侵袭和转移的影响朱军，高娟，李红彦,潘湘阳，陈俊霞(400016重庆，重庆医科大学细胞生物学与遗传学教研室，分子医学与肿瘤研究中心)[摘要]目的设计合成有效的ILK-siRNA,并检测其对EJ细胞侵袭和迁移的影响。方法构建针对ILK基因mRNA的2个特异性siRNA表达载体pGenSil-1siRNA1、pGensil-1siRNA2和1个阴性对照载体pGenSil-1siRNA3。稳定转染膀胱癌EJ细胞后，通过RT-PCR检测EJ细胞中ILKmRNA水平的表达，并通过Westernblot和细胞免疫荧光检测ILK
2、蛋白水平的表达；观察EJ细胞形态变化；检测EJ细胞迁移、黏附和侵袭能力及基质金属蛋白酶的变化。结果重组质粒构建成功。干扰组细胞ILKmRNA及蛋白的表达分别较未转染组、转空质粒组和转阴性对照质粒组相比，呈显著性下降（P<0.05）；HE染色结果显示，细胞铺展良好，核质比减小；转染干扰质粒的2个实验组细胞的黏附能力相对于对照组分别增加87%和76%，黏附能力显著升高，迁移及侵袭能力明显降低（P<0.05）；与其他3个对照组相比,转染pGenSil-1siRNA1与pGensil-1siRNA2质粒到EJ细胞后,MMP-2及MMP-9的表达水平明显降低（P<
3、0.05）。结论成功构建的干扰质粒能抑制ILK基因及蛋白水平的表达，显著抑制细胞的生长、侵袭及迁移能力。[关键词]ILK;siRNA；膀胱癌EJ细胞；侵袭；迁移ILKSilencingintegrin-linkedkinasebySmallsmallInterferinginterferingRNAInhibitsinhibitsMetastasismetastasisandInvasioninvasionBehaviourofHumanhumanEJBladderbladderCancercancerCellcellsZhuJun,GaoJuan,Li
4、Hongyan,PanXiangyang,ChenJunxia（(DepartmentofCellbBiologyandGeneticsDivision,CenterofMolecularMedicineandCancerResearchCenter，,CollegeofBasicMedicalSciences,ChongqingMedicalUniversity,Chongqing,400016,China);[Abstract]ObjectiveToinvestigatetheeffectofthespecificsiRNAofintegrin-li
5、nkedkinase(ILK)onthemigrationandinvasionofhumanEJbladdercancercells.MethodsConstructtTwospecificsiRNAexpressionvectorsandonenonehomologycontrolvectortargetingtheobjectivegenewasconstructed.TransfecttThenthevectorsweretransfectedintoEJcellsrespectively.DetecttheRNAiefficiencyonthe
6、expressionofILKwasevaluatedbyRT-PCR,Western-blottingandIimmunofluorescencestaining.ObservetThemorphouslogicalchange,celldeterminetheadhesion,migrationandinvasionability,andtheexpressionsofMMP2andMMP9ofthetransfectedcellsweredetected.ResultsEnzymedigestionandDNAsequencingverifiedt
7、heconstructionedvectorswerecorrectly.Inthetwo2specificsiRNAtreatmentgroups,theexpressionofILKwasobviouslyinhibited,thecellsspreadwell,thenuclearmassratiowaslowerthancontrolgroups,thecelladhesionabilitywasincreased,thecellmigrationandmetastasisabilityweredecreased,andtheexpression
8、ofMMP2andMMP9obviouslydecreased.Conclusi
