microrna-149对膀胱癌t24细胞体外增殖、凋亡及侵袭的影响word论文.docxVIP

microRNA-149*对膀胱癌T24细胞体外增殖、凋亡及侵袭的影响中文摘要目的：观察microRNA-149*（miR-149*）对人膀胱癌T24细胞体外增殖、凋亡、和侵袭的影响，探讨miR-149*对膀胱癌T24细胞发生发展的可能机制。方法：应用化学方法合成microRNA-149* mimics，瞬时转染T24细胞。实时定量RT-PCR，WesternBlot实验分别从基因和蛋白方面验证转染后microRNA-149* 在T24细胞中的表达上调。然后分别通过流式细胞计数、MTT、transwell实验、细胞粘附实验分别检测microRNA-149* 对T24细胞的凋亡、增殖、侵袭能力及细胞粘附能力的影响。结果：1.化学合成microRNA-149*mimics，瞬时转染T24细胞，浓度在50nM时miRNA转染细胞率在90%。2.实时荧光定量PCR 及Western blot实验，结果miR-149*的表达在转染miR-149*mimics后的T24细胞组中与阴性对照组及空白对照组相比明显增加，表明miR-149*在膀胱癌T24细胞中明显上调。3.流式细胞学检测，结果转染miR-149*mimics后的T24细胞其凋亡和坏死比例明显上升。4.MTT实验细胞计数后，结果表明转染miR-149*后的T24细胞生长速度减慢，细胞增殖受抑制。5.transwell实验穿过基质胶的细胞计数，结果表明转染miR-149*后的T24细胞体外侵袭能力减弱。6.细胞粘附实验结果所示，转染miR-149*后的T24细胞粘附性增强。结论：1.体外转染miR-149*后的T24细胞增殖受抑制、加快其凋亡、细胞粘附性增加，并且降低其侵袭能力。2.miR-149*差异表达可能参与了人膀胱移行细胞癌的发生与发展。关键词：microRNA-149*；细胞增殖；凋亡；侵袭1EffectsofmiR-149* onproliferation,apoptosis and invasion of bladdercancerT24 cells in vitroAbstractObjiec:Toobsivetheproliferation,invasionandapotosisinfluenceofmiR-149*for bladder cancer cellsT24 occurrence and development ofthe possiblemechanism. Methods:Real-time quantitative RT-PCR,Western Blot T24 cells showed low expression proteins in microRNA-149* in the T24 cells whether differences in expression.ApplicationchemicalsynthesisofmicroRNA-149*mimics,transientrans fection ofT24 cells,respectively by flowcytometry,MTT,transwell experiments; celladhesion expermiments were used to detect microRNA-149* on T24 cell apoptosis,proliferation,invasion and cell adhesion capacity.Results:1.sytheticmicroRNA-149*mimicstransfectionofT24cells,the concentrationof50NmmiRNAtransfectedcellswhentherateof90%.2.Real-time PCRandWesternblotexperiments,theresultsofmiR-149*expressionintransfected miR-149*mimicsthegroupafterT24cellsandT24cells,emptyvectorgroupand significantlythemigrationofinvasion;miR-149*maybeincreasedcomparedT24 cells,indicatingthatmiR-149*T24bladdercancercellsinthedifferential expression.3.flowcytometry,andtheresultstransfectedandreducethemigrationof invasionwithmiR-149*mimicstheT24cellsaf