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1、依布硒诱导多发性骨髓瘤细胞凋亡及其机制的研究张亮,杜佳,周立为,程燚,王东,谢家印(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]目的探讨依布硒对多发性骨髓瘤细胞增殖、凋亡的影响及机制。方法应用CCK8观察不同浓度依布硒作用于人多发性骨髓瘤细胞株U266细胞8、24、48、72h后细胞活力及增殖情况,同时应用Hoechst染色检测细胞凋亡,JC-1检测细胞线粒体膜电位,ROS探针观察依布硒对细胞内活性氧的影响,以及Westernblot检测Bcl-2、Bax、细胞色素C等凋亡相关蛋白及Caspase9激活情况。结果依布硒能够显著抑制多发性骨髓瘤细胞增殖,并
2、诱导细胞凋亡,40μmol/L依布硒使凋亡率达(80.8±3.9)%;同时依布硒能升高细胞内ROS水平,降低线粒体膜电位;此外依布硒可诱导细胞色素C从线粒体向胞浆释放但并不激活Caspase9。结论依布硒可诱导多发性骨髓瘤细胞发生非Caspase依赖的细胞凋亡,其机制可能与ROS水平升高有关。[关键词]依布硒;多发性骨髓瘤;凋亡;活性氧[中图法分类号][文献标志码]AEbseleninducesapoptosisinhumanmultiplemyelomacelllinesanditsmechanismZhangLiang,DuJia,ZhouLiwei,ChengYi,Wang
3、Dong,XieJiayin(Cancercenter,DapingHospitalandInstituteofSurgeryResearch,ThirdMilitaryMedicalUniversity,Chongqing,4000042,China)[Abstract]ObjectiveToinvestigatetheeffectofEbselenoncellproliferationandapoptosisinmultiplemyeloma(MM)cellsanditsprobablymechanism.MethodsCellviabilityofhumanMMcellli
4、neU266wasexaminedbyCCK8assaykit.Theproductionofreactiveoxygenspecies(ROS)wasmeasuredbySingletROSProbe.JC-1wasusedtoassaythemitochondrialmembranepotential(△Ψm).TheapoptosisassaywasdonethroughHoechststaining.TheproteinlevelsofBcl-2,Bax,caspase9andcytochromecweredetectedbyWesternblot.ResultsEbse
5、lenwasfoundtosignificantlydecreasecellviabilityandincreaseapoptosisaccompaniedbyenhancingproductionofROSinU266cells.Furtherstudiesrevealedcytochromecreleasefromthemitochondriatocytosolclinesaccompaniedbymitochondrialmembranepotential△Ψmdecrease,whilecaspase9wasnotobservedtobeactivated.Conclus
6、ionEbselenmayenhancetheproductionofendogenousROS,andinduceMMcellapoptosisviamitochondria-mediatedapoptoticpathway.[Keywords]EbselenMultipleMyelomaApoptosisROSEbseleninduceapoptosisinhumanmultiplemyelomacelllinesanditsmechanismZhangLiang,DuJia,ZhouLiwei,ChengYi,WangDong,XieJiayin(Cancercenter,
7、DapingHospitalandInstituteofSurgeryResearch,ThirdMilitaryMedicalUniversity,Chongqing,4000042,China)[Abstract]Objective:ToinvestigatetheeffectofEbselenoncellproliferationandapoptosisinMultipleMyeloma(MM)cellsanditsprobablymechanism.Methods:The