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《rna干扰骨髓基质细胞和或骨髓瘤细胞ape1表达对共培养骨髓瘤细胞增殖及凋亡的影响》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、RNA干扰骨髓基质细胞和/或骨髓瘤细胞APE1表达对共培养骨髓瘤细胞增殖及凋亡的影响谢家印,杜佳,李梦侠,卿毅,关伟,曾林立,王东(400042重庆,第三军医大学大坪医院野战外科研究所肿瘤中心)[摘要]目的探讨RNA干扰骨髓基质细胞(bonemarrowstromalcells,BMSCs)及骨髓瘤细胞株U266APE1表达对共培养的U266细胞增殖、凋亡的影响。方法将构建的APE1siRNA表达载体分别导入BMSCs及U266细胞中。Westernblot法检测2种细胞中APE1蛋白表达;2株细胞经APE1siRNA处理后通过Transwel
2、l插入式培养皿构建骨髓基质细胞与骨髓瘤细胞共培养模型,采用MTT法、AnnexinV-PE/7-AAD双染法、RT-PCR分别检测U266细胞增殖、凋亡及细胞中IL-6/IL-8mRNA表达水平。结果结果应详尽一些。补充P值APE1siRNA可明显降低骨髓基质细胞及U266细APE1蛋白表达,与对照组相比差异具有统计学意义(P<0.01);在APE1siRNA同时处理BMSCs和U266细胞后的共培养体系中,U266细胞增殖抑制及细胞凋亡明显高于单一细胞APE1敲低组及APE1siRNA未处理组(P<0.01);U266细胞中IL-6及IL-8
3、mRNA表达水平亦降低(P<0.01)。结论阻抑APE1在骨髓基质细胞和/或骨髓瘤细胞株U266的表达,可明显抑制共培养体系中U266细胞的增殖活性,并促进其凋亡。[关键词]多发性骨髓瘤;细胞凋亡;APE1基因;RNA干扰[中图法分类号][文献标志码]AEffetsofRNAinterferenceinhibitingAPE1expressioninbonemarrowstromalcellsandU266onproliferationandapoptosisofco-culturedU266XieJiayin,DuJia,LiMengxia,
4、QingYi,GuanWei,ZengLinli,WangDong(CancerCenter,InstituteofSurgeryResearch,DapingHospital,ThirdMilitaryMedicalUniversity,Chongqing,400042,China)[Abstract]对照中文修改ObjectiveTostudytheeffectofRNAinterferenceinhibitingapurinic/apyrimidinicendonuclease(APE1)expressioninbonemarrowstr
5、omalcells(BMSCs)andU266onproliferationandapoptosisofco-culturedU266.MethodsAPE1siRNAexpressionvectorweretransfectedtoBMSCsandU266cells,theexpressionlevelofAPE1wasdetectedbyWesternblot;Co-culturesystemofBMSCsandU266cellswasestablishedbyTranswellinsertculturedishafterthetransf
6、ectionoftheadenoviralvectorAd5-APE1siRNA.TheproliferationofU266cellsweredeterminedbyMTTmethod,thecellapoptosisratebyflowcytometrywithAnnexinV-PE/7-AADdoublestaining,theexpressionlevelsofIL-6/IL-8mRNAbyRT-PCR.ResultsTheexpressionofAPE1proteindecreasedsignificantlyinBMSCsandU2
7、66cellsbyRNAinterferingcomparedwithcontrolgroup(P<0.01).Theco-culturesystemofBMSCsandU266cellsbyRNAinterfering,theeffectofinhibitiononproliferationandapoptosisofU266cellswereincreasedsignificantly(P<0.01),theexpressionlevelsofIL-6/IL-8mRNAweredecreased(P<0.01).ConclusionThei
8、nhibitionofAPE1expressioninBMSCsand/orU266cellsinhIbitstheproliferationandp