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1、窯764窯中華神經(jīng)醫(yī)學(xué)雜志2011年8月第10卷第8期ChinJNeuromed,August2011,Vol.10,No.8窯基礎(chǔ)研究窯人腦膠質(zhì)瘤多藥耐藥細(xì)胞株的構(gòu)建及生物學(xué)特性鑒定白義鳳廖紅展劉天助郭洪波揖摘要銥?zāi)康捏w外構(gòu)建膠質(zhì)瘤耐甲磺酸伊瑪替尼多藥耐藥細(xì)胞株袁并研究其生物學(xué)特性遙方法采用逐漸增加培養(yǎng)基中伊瑪替尼藥物濃度的方法誘導(dǎo)構(gòu)建耐甲磺酸伊瑪替尼的人腦膠質(zhì)瘤U251細(xì)胞株(命名為U251AR)曰CCK8法檢測(cè)U251AR堯U251對(duì)多種化療藥物的IC50和耐藥指數(shù)曰QRT-PCR法檢測(cè)耐藥相關(guān)基因ABCC1堯ABC
2、B1堯ABCB4堯ABCG2mRNA的表達(dá)袁流式細(xì)胞術(shù)檢測(cè)ABCG2蛋白的表達(dá)遙結(jié)果體外培養(yǎng)12個(gè)月后建立了穩(wěn)定耐藥的細(xì)胞株U251AR袁與親代細(xì)胞相比異形性不顯著遙U251和U251AR對(duì)化療藥物的IC50相比較差異均有統(tǒng)計(jì)學(xué)意義(P<0.05)袁U251AR對(duì)甲磺酸伊瑪替尼堯阿霉素堯順鉑的耐藥指數(shù)分別為20.41堯5.06堯10.28袁表現(xiàn)出多藥耐藥特征遙QRT-PCR檢測(cè)結(jié)果顯示耐藥細(xì)胞株U251AR中ABCC1堯ABCB1堯ABCB4堯ABCG2mRNA的表達(dá)明顯高于親代U251細(xì)胞袁流式細(xì)胞術(shù)檢測(cè)結(jié)果顯示U251
3、AR中ABCG2蛋白的表達(dá)強(qiáng)于親代U251細(xì)胞袁差異均有統(tǒng)計(jì)學(xué)意義(P<0.05)遙結(jié)論成功建立多藥耐藥的膠質(zhì)瘤細(xì)胞株U251AR袁其多藥耐藥與ABCC1堯ABCB1堯ABCB4堯ABCG2mRNA及ABCG2蛋白的表達(dá)上調(diào)相關(guān)遙揖關(guān)鍵詞銥神經(jīng)膠質(zhì)瘤曰甲磺酸伊瑪替尼曰多藥耐藥性曰細(xì)胞株揖中圖分類號(hào)銥R730.264揖文獻(xiàn)標(biāo)志碼銥A揖文章編號(hào)銥1671-8925(2011)08-764-04Establishmentofhumanglioblastomamultiformmultidrugresistantcelllinei
4、nvitroandidentificationofitsbiologicalcharacteristicsBAIYi-feng*,LIAOHong-zhan,LIUTian-zhu,GUOHong-bo.*DepartmentofOncology,ZhujiangHospital,SouthernMedicalUniversity,Guangzhou510282,ChinaCorrespondingauthor:GUOHong-bo,Email:hongboguo158@126.com揖Abstract銥Objective
5、Toestablishtheimatinib(STI-571)-resistantsublineinvitroandinvestigateitsbiologicalcharacteristics.MethodsHumanglioblastomamultiformdrug-resistantcellline(namedU251AR)wasestablishedinvitrobysuccessivelyincreasingtheconcentrationofimatinibinacellculturemedium.The50%
6、inhibitorydose(IC50)valuesandtheresistanceindexes([IC50U251/STI-571]/[IC50U251])forotherchemotherapeuticagentswereevaluatedusingcellcountingkit-8assays.ExpressionsofacquiredmultidrugresistanceP-glycoprotein(MDR1,ABCB1;MDR3,ABCB4),breastcancerresistanceprotein(BCRP
7、,ABCG2)andmultidrugresistance-associatedprotein1(MRP1,ABCC1)weredetectedbyQRT-PCR.FlowcytometrywasemployedtodetecttheproteinexpressionofABCG2.ResultsTheU251ARwasdevelopedafterculturefor12monthsandsimilarmorphologiesofU251andU251/STI-571cellsweredetermined.Theresis
8、tancecoefficientofU251ARcellstoimatinibwas20.41timesmorethanthatoftheparentcells,andU251ARcellsshowedcross-resistancetomanyanti-tumoragents(P<0.05).Ther
