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《Gankyrin基因沉默對(duì)人卵巢癌SKOV3/CDDP細(xì)胞順鉑耐藥性的逆轉(zhuǎn)作用和機(jī)制-論文.pdf》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫。
1、中國(guó)癌癥雜志2014年第24卷第1期CHINAONCOLOGY2O14Vo1.24No.135Gankyrin基因沉默對(duì)人卵巢癌SKOV3/CDDP~胞順鉑耐藥性的逆轉(zhuǎn)作用和機(jī)制王倩程墊天津市水閣醫(yī)院婦產(chǎn)科,天津300120[摘要]背景與目的:卵巢癌是常見的婦科腫瘤,抗腫瘤藥耐藥性的產(chǎn)生是卵巢癌治療失敗的主要原因之一,Gankyrin基因被認(rèn)為與腫瘤耐藥性密切相關(guān),本文探討了Gankyrin基因沉默對(duì)卵巢癌耐順鉑細(xì)胞系SKOV3/DDPJI[~鉑耐藥性的逆轉(zhuǎn)作用及機(jī)制。方法:應(yīng)用real—timePCR技術(shù)考察Gankyrin在
2、SKOV3~NSKOV3/DDPfm胞中的表達(dá),應(yīng)用MTS法檢~6ankyrin對(duì)SKOV3/DDP細(xì)胞順鉑耐受性的影響,應(yīng)用流式細(xì)胞術(shù)檢測(cè)腫瘤細(xì)胞凋亡和細(xì)胞內(nèi)羅丹明一123(Rhodamine—I23,Rh一123)含量的變化,Westernblot和rea卜timePCR技術(shù)檢測(cè)腫瘤細(xì)胞耐藥相關(guān)蛋白MDR1、Caspase3/8、SurvivinUBc卜2蛋白表達(dá),Westernb1ot法檢~lJp53、NF—KB和PTEN蛋白表達(dá)UAKT磷酸化水平。結(jié)果:Gankyrin在SK0V3/DDP細(xì)胞中表達(dá)升高,沉默Gankyr
3、in基因后可增加SKOV3/DDP細(xì)胞對(duì)順鉑的敏感性?;虺聊昂竽退幠孓D(zhuǎn)倍數(shù)(resistantfactor,RF)為i.81和2.45,腫瘤細(xì)胞中Rh一123含量提高了1.73和2.42倍,細(xì)胞凋亡率是對(duì)照組的2.23倍和4.23倍,耐藥相關(guān)蛋白MDR1、Survivin和Bc卜2蛋白水平顯著下降,MDR1mRNA表達(dá)是對(duì)照組的62.8%和21.6%,SurvivinmRNA表達(dá)是對(duì)照組的24.5%和i0.3%,Bc卜2mRNA表達(dá)是對(duì)照組的47.5%和18.4%,Caspase一3/8、P53和PTEN表達(dá)水平上升,AKT
4、磷酸化和NF—KB水平下降。結(jié)論:沉默Gankyrin基因可逆轉(zhuǎn)SKOV3/DDP對(duì)順鉑的耐藥性,可能與抑制藥物外排,促進(jìn)細(xì)胞凋亡有關(guān),PTEN/AKT/NF—KB/p53信號(hào)通路可能是其中心環(huán)節(jié)。[關(guān)鍵詞]Gankyrin基因;卵巢癌;順鉑耐藥DOI:10.3969/j.issn.1007—3969.2014.01.006中圖分類號(hào):R737_31文獻(xiàn)標(biāo)志碼:A文章編號(hào):1007—3639(2014)01.0035.06TheeffectsandmechanismsofGankyrinsilencingonreversingt
5、hecisplatinresistanceofhumanovariancancerSKOV3/DDPcelllineWANGQian,CHENGKun(DepartmentofGynaecologyandObstetrics,TianjinShuiGeHospital,Tianjin300120,China)Correspondenceto:WANGQianE—mail:wangqiansgh@126.corn[Abstract]Backgroundandpurpose:Ovariancanceristhecommongynec
6、ologicalcancer,andthedrugresistanceofanti—tumordrugwasoneofmajorreasonsfortherapyfailure,somestudiesconsideredthatthereisaclosedrelationshipbetweenGan@rinanddrugresistance.Inthisstudy,weinvestigatedtheefectsandmechanismsofGankyrinsilencingonreversingthecisplatinresis
7、tanceofovariancancerdrug—resistantSKOV3/DDPcellline.Methods:TheexpressionofOankyrininSKOV3andSKOV3/DDPcellswasmeasuredbyrea1.timePCRassay,MTSassaywasemployedtodeterminetheeffectofGankyrinonSKOV3/DDPsensitivitytocisplatin,apoptosisrateandintracellularconcentrationofrh
8、odamine.123rRh一123)weredeterminedbyflowcytometry,theexpressionofmulti.drugsresistantproteinMDR1Caspase一3/8.SurvivinandBcl一2weredete