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1、廣東醫(yī)學(xué)2014年5月第35卷第9期GuangdongMedicalJournalMay.2014,Vo1.35,No.9·1317·冰片對(duì)Ecal09/DDP細(xì)胞耐順鉑的逆轉(zhuǎn)作用米戴翠萍,張徐寧,薛彩萍,季寧東淮陰衛(wèi)生高等職業(yè)技術(shù)學(xué)?;窗彩邢滥[瘤重點(diǎn)實(shí)驗(yàn)室(江蘇淮安223300)【摘要】目的觀察冰片對(duì)食管癌細(xì)胞Ecal09/DDP耐順鉑(DDP)的逆轉(zhuǎn)作用,并探討其機(jī)制。方法MTT法檢測(cè)冰片單用及與DDP聯(lián)用對(duì)Ecal09/DDP細(xì)胞增殖的影響,觀察冰片對(duì)Ecal09/DDP細(xì)胞耐DDP的逆轉(zhuǎn)作用,計(jì)算耐藥逆轉(zhuǎn)指數(shù);Westernblot法檢測(cè)Ecal09/DDP細(xì)胞P一糖蛋白(
2、P—gP)的表達(dá)水平。結(jié)果0.195、0.390、0.780、1.560g/mL冰片對(duì)Ecal09/DDP細(xì)胞無(wú)明顯毒性作用(P>0.05),但與lmL的DDP聯(lián)用能濃度依賴性增加DDP對(duì)Ecal09/DDP細(xì)胞的毒性(P<0.05),降低DDP對(duì)Ecal09/DDP細(xì)胞的半數(shù)抑制濃度(Ic)(P<0.05),耐藥逆轉(zhuǎn)指數(shù)高達(dá)5.31,并能下調(diào)Ecal09/DDP細(xì)胞P—gP的表達(dá)水平(P<0.05)。結(jié)論冰片能逆轉(zhuǎn)Ecal09/DDP細(xì)胞對(duì)DDP的耐藥,其作用與下調(diào)Ecal09/DDP細(xì)胞P—gp的表達(dá)有關(guān)?!娟P(guān)鍵詞】冰片;順鉑;Ecal09/DDP;耐藥逆轉(zhuǎn);P一糖蛋白Therev
3、ersalefectofborneoloncisplatinresistanceinEcal09/DDPcells.DAICui—ping,ZHANGXu—ning,XUECai—ping,JlⅣ一.KeyLaboratoryofTumorinDigestiveSystemofHuaian,HuaiyinAdvancedVocationalandTechnicalHealthSchool,Huaian223300,ChinaCorrespondingauthor:JlM一如.E—mail:hywxjndl@163.con【Abstract】ObjectiveTostudythereve
4、rsaleffectandmechanismofborneoloncisplatin(DDP)resistanceinhumanesophagealcarcinomacellEcal09/DDP.MethodsThecytotoxiceffectofborneolorDDPonEcal09/DDPcellswasmeasuredbyMTrandthedrug—resistantreversalindex(RI)wascalculated.TheexpressionofP—glycoprotein(P—gP)wasdetectedbyWesternblot.ResultsMTFassay
5、showedthatborneolattheconcentrationnomorethan1.560g/mLhadnoobviouscytotoxicitytoEcal09/DDPcells(P>0.05),butitcouldsignificantlyenhancethecytotoxicityofDDP(1mL)toEcal09/DDPcellsinaconcentration—dependentmanner(P<0.05),reduceIC50ofDDPonEcal09/DDPcells(P<0.05),anddown—regulatetheexpressionofP—gpofE
6、cal09/DDPcells(P<0.05).RIofDDPwas5.31.ConclusionBorneolpossessesareversaleffectonDDPresistanceinEcal09/DDPcells.Themechanismmaybeassociatedwiththedown—regulationofP—glycoprotein.【Keywords】borneol;cisplatin;Ecal09/DDP;reverseofdrugresistance;P—glycoprotein冰片為龍腦香科植物龍腦香樹(shù)脂的加工品或菊科白定量試劑盒(南京凱基生物科技發(fā)展有限公
7、司),植物艾納香葉提取的結(jié)晶,或以樟腦、松節(jié)油等為原P一糖蛋白(P—glycoprotein,P—gP)單克隆抗體(美國(guó)料,經(jīng)化學(xué)方法合成的精制品?。冰片的藥理作用廣Abcam公司),兔甘油醛一3一磷酸脫氫酶(GAPDH)多泛,但對(duì)腫瘤細(xì)胞作用的研究很少。2010年5月至克隆抗體、辣根過(guò)氧化物酶標(biāo)記的二抗(美國(guó)Santa2013年4月,本研究以耐順鉑(cisplatin,DDP)的食管Cruz公司),其他試劑均為國(guó)產(chǎn)分析純。癌細(xì)胞Ecal09/