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1、醫(yī)藥導(dǎo)報(bào)2014年9月第33卷第9期·1121··藥物研究·替莫唑胺耐藥人腦膠質(zhì)瘤細(xì)胞系*U251/TR建立及耐藥機(jī)制12345潘強(qiáng),楊學(xué)軍,高松,紀(jì)延偉,張文高(1.山東省萊蕪市人民醫(yī)院、泰山醫(yī)學(xué)院附屬萊蕪醫(yī)院神經(jīng)外科,萊蕪搖271100;2.天津醫(yī)科大學(xué)總醫(yī)院神經(jīng)外科,天津搖300052;3.天津醫(yī)科大學(xué)附屬腫瘤醫(yī)院胰腺腫瘤科,天津搖300060;4.山東省交通醫(yī)院神經(jīng)外科,濟(jì)南搖250031;5.河北省滄州市中心醫(yī)院神經(jīng)外科,滄州搖061001)摘搖要搖目的搖建立替莫唑胺耐藥人腦膠質(zhì)瘤細(xì)胞系,探討
2、其耐藥機(jī)制,以期為臨床優(yōu)化藥物化療方案提供參考。方法搖通過體外分步誘導(dǎo)方法使人U251膠質(zhì)瘤細(xì)胞對(duì)替莫唑胺產(chǎn)生耐藥,噻唑藍(lán)比色法檢測(cè)耐藥指數(shù)及細(xì)胞存活率;6采用Western鄄Blot、逆轉(zhuǎn)錄鄄聚合酶鏈免疫(RT鄄PCR)、免疫熒光、免疫組化法檢測(cè)O鄄甲基鳥嘌呤鄄DNA甲基轉(zhuǎn)移酶(MGMT)表達(dá)變化,分析細(xì)胞耐藥機(jī)制。結(jié)果搖歷經(jīng)8個(gè)月成功建立了對(duì)替莫唑胺耐藥的膠質(zhì)瘤細(xì)胞U251/TR,在替莫-1-1唑胺初始濃度為0.25滋g·mL,終濃度為16.00滋g·mL培養(yǎng)液中,U251/TR細(xì)胞半數(shù)抑制濃度(I
3、C)為未經(jīng)誘導(dǎo)的50U251膠質(zhì)瘤細(xì)胞的7倍(P=0.00),其耐藥指數(shù)約為7.00;U251/TR細(xì)胞MGMT表達(dá)水平較未經(jīng)誘導(dǎo)的U251膠質(zhì)瘤細(xì)胞明顯增加(P=0.00)。結(jié)論搖通過分步誘導(dǎo)方法于體外成功建立1株對(duì)替莫唑胺耐藥的U251/TR細(xì)胞系。MGMT表達(dá)水平升高是導(dǎo)致U251/TR細(xì)胞對(duì)替莫唑胺耐藥的主要機(jī)制。6關(guān)鍵詞搖替莫唑胺;神經(jīng)膠質(zhì)瘤;O鄄甲基鳥嘌呤鄄DNA甲基轉(zhuǎn)移酶;抗藥性中圖分類號(hào)搖R979.1;R969搖搖搖文獻(xiàn)標(biāo)識(shí)碼搖A搖搖搖文章編號(hào)搖1004-0781(2014)09-112
4、1-05EstablishmentofaTMZ鄄resistantHumanGliomaCellLineU251/TRandtheMechanismofDrug鄄resistance12345PANQiang,YANGXue鄄jun,GaoSong,JIYan鄄wei,ZHANGWen鄄gao(1.DepartmentofNeurosurgery,People'sHospitalofLaiwuCity,LaiwuHospitalAffilliatedtoTaishanMedicalCollege,Lai
5、wu271100,China;2.DepartmentofNeurosurgery,TianjinMedicalUniversityGeneralHospital,Tianjin300052,China;3.DepartmentofPancreaticNeoplasms,CancerHospital,TianjinMedicalUniversity,Tianjin300060,China;4.DepartmentofNeurosurgery,ShandongJiaotongHospital,Jinan2
6、50031,China;5.DepartmentofNeurosurgery,CangzhouCentralHospitalofHebeiProvince,Cangzhou061001,China)ABSTRACT搖Objective搖Toestablishadrug鄄resistancecelllineofhumangliomawithtemozolomide(TMZ),investigateitsresistancemechanisms,andprovideexperimentalevidencef
7、oroptimalTMZtherapy.搖Methods搖ATMZ鄄resistanthumangliomacellline,U251/TR,wasestablishedbystepwiseexposureofhumanparentalU251cellstoTMZ.ResistanceindexandcellviabilitywereaccessedbyMTTassay.Western鄄Blot,RT鄄PCR,immunohistochemistryandimmunofluorescencewereus
8、edtodetectMGMTexpressionfortheanalysisofresistancemechanism.搖Results搖ATMZ鄄resistanthumangliomacellline,U251/TR,was-1developedafter8monthsofstepwiseinductionwith0.25-16.00滋g·mLTMZ.ICinU251/TRcellswasapproximately750timeshig
