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1、··中國肺癌雜志2010年1月第13卷第1期ChinJLungCancer,January2010,Vol.13,No.1·基礎(chǔ)研究·GRP78在肺腺癌細(xì)胞順鉑耐藥中的作用吳晶王琪王佳瑞張黎川趙龍【摘要】背景與目的糖調(diào)節(jié)蛋白78（GRP78）在肺癌細(xì)胞中表達(dá)增高并與其對化療藥物VP-16的耐藥性有關(guān)，而GRP78的表達(dá)與其對化療藥物順鉑的耐藥性是否相關(guān)，研究較少。本研究旨在通過檢測GRP78在人肺腺癌細(xì)胞SPCA-1中的表達(dá)及順鉑作用后的細(xì)胞生存率，研究GRP78在肺腺癌細(xì)胞順鉑耐藥中的作用。方法依
2、據(jù)是否應(yīng)用誘導(dǎo)劑A23187將細(xì)胞分為實(shí)驗(yàn)組和對照組，采用RT-PCR、Westernblot方法檢測各組細(xì)胞中GRP78在核酸和蛋白水平的表達(dá)，利用MTT法測定細(xì)胞在順鉑作用下的生存率，分析GRP78在SPCA-1細(xì)胞順鉑耐藥中的作用。結(jié)果A23187可以明顯誘導(dǎo)SPCA-1細(xì)胞中GRP78在核酸和蛋白水平的表達(dá)，且其表達(dá)水平與A23187具有一定的濃度依賴性，MTT檢測顯示：實(shí)驗(yàn)組細(xì)胞生存率明顯低于對照組，亦呈一定的A23187濃度依賴性。結(jié)論在人肺腺癌細(xì)胞SPCA-1中，誘導(dǎo)劑A23187可以誘導(dǎo)
3、GRP78核酸和蛋白水平的表達(dá)，其表達(dá)與肺癌細(xì)胞在順鉑作用下的生存率呈負(fù)相關(guān)，表明GRP78可提高SPCA-1細(xì)胞對順鉑的敏感性，與肺癌細(xì)胞化療耐藥有一定的相關(guān)性?！娟P(guān)鍵詞】葡萄糖調(diào)節(jié)蛋白78（GRP78）；肺腫瘤；順鉑；耐藥性【中圖分類號】R734.2DOI:10.3779/j.issn.1009-3419.2010.01.07TheRoleofGRP78ontheResistancetoCisplatininSPCA-1CellLine12222JingWU,QiWANG,JiaruiWANG,Li
4、chuanZHANG,LongZHAO12DepartmentofRespiratoryMedicine,theDalianNo.4PeopleHospital,Dalian116031,China;DepartmentofRespiratoryMedicine,theSecondHospitalofDalianMedicalUniversity,Dalian116031,ChinaCorrespondingauthor:QiWANG,E-mail:wqdmu@yahoo.com.cn【Abstract
5、】BackgroundandobjectiveTheexpressionlevelsofglucose-regulatedprotein78(GRP78)wereelevatedandcorrelatedwithresistancetochemotherapydrugVP-16inlungcancercells.However,littleisknownabouttherelationshipbetweenitsexpressionandresistancetocisplatininlungcancer
6、cells.TheaimofthisstudywastoinvestigatetheexpressionofGRP78undertheinductionofA23187anditssignificanceintheresistancetoanti-tumordrugscisplatininahumanlungcancerSPCA-1cellline.MethodsRT-PCRandWesternblotwereusedtoanalyzetheexpressionofGRP78atmRNAandprote
7、inlevelsinSPCA-1celllineinducedbyA23187atdifferentconcentrations(0,1,2,4,6μM).MTTwasusedtodeterminetheeffectofcisplationoncellsurvival.ResultsTheexpressionsofGRP78atbothmRNAandproteinlevelswereincreasedobviouslyinSPCA-1celllineinducedbyA23187,withamanner
8、ofdose-dependentofA23187toagreatdegree;MTTassayshowedthatthecellsurvivalrateoftheA23187-inducedgroupdecreasedsignificantlycomparetothecontrolgroup,alsowithaconcentration-dependentmannerofA23187.ConclusionTheexpressionofGRP
