卵巢癌细胞对顺铂耐药机制的研究.doc

卵巢癌细胞对顺铂耐药机制的研究
癌症1999年第6期第19卷基础研究
作者：陈葳，李旭，杨玉琮，程小丽，牛映斗
单位：西安医科大学第一附属医院临床分子生物学中心(西安，710061)
关键词：卵巢癌细胞株；顺铂；药物耐受性
【摘要】目的：探明卵巢癌对顺铂产生耐药的机理。方法：采用顺铂体外诱导法建立卵巢癌耐药细胞株HO-8910/2。MTT法测定其耐药倍数和交叉耐药性，原子吸收法测定细胞内Pt浓度，分光光度法测定GSH、GST，流式细胞仪分析细胞周期，检测bcl-2和P-GP的表达。结果：HO-8910/2耐顺铂是亲代细胞HO-8910的6.6倍，与5-FU，ADR有交叉耐药性。对照亲代细胞，耐药细胞中GSH含量与GST活性明显增高，细胞G2/M比例增加，细胞内铂(Pt)摄入和蓄积减少，bcl-2表达阳性细胞增多，P-GP无变化。结论：细胞内药物浓度下降、bcl-2的表达增加、GSH与GST对铂化合物解毒作用的增强可能是HO-8910/2耐药机制形成的主要原因。
中图分类号：R737.31文献标识码：A文章编号：1000－467X(1999)06－0674－03
Experimentalstudyonthemechanismofcisplatinresistanceinhuman
ovariancarcinomacells
CHENWei，LIXu，YANGYu-cong，etal.
CenterforClinicalMolecularBiology，theFirstAffiliatedHospitalofXi’anMedicalUniversity.Xi’an710061，P.R.China
【Abstrct】Objective：Toexplorethemechanismofcisplatinresistanceinhumanovariancancer.Methods：Acisplatinresistanthumanovariancancercellline，HO-8910/2，wasdevelopedinvitroafterprolongeddrugexposureofthecisplatin-sensitiveparentalHO-8910cellline.Drugcytotoxicitiesweredeterminedusingthemicrotetrazoliumassay.TheconcentrationsofcellularPtweredeterminedbyatomicabsorptionandthelevelsofcellularGSHandGSTbyweredetectedspectrophotometry.Cellscycleandexpressionofbcl-2andP-gpwereanalysedbyflowcytometry.Results：HO-8910/2celllineexhibitedadegreeofresistanceofapproximately6.6timescomparedwiththeparentalcelllinefollowing48hexposuretothedrugsandwascross-resisanceto5-FUandVCR.A44.1％reductionwasobservedinplatinumaccumulationintheselectedcellsafterculturingwithCDDPfortwohours.IncreasedlevelsofcellularglutathioneandglutathioneS-transferasewerefoundinthecisplatin-resistantcellswiththeresultsofGSH(μmol/ml)17.63±2.00vs15.63±1.70(P＜0.05)andGST(IU/ml)9.67±2.05vs5.33±1.70(P＜0.01)，respectively.HO-8910/2cellsrevealedahighpercentofG2/Mandanoverexpressedbcl-2protein(75.9％vs34.4％)，buttherewasnodifferenceinP-gpexpressionbetweenthetwocelllines.Conclusins：Itisdemonstratedthatdecreaseddrugaccumulation，reducedsusceptibilitytocisplatin-inducedapoptosisandincreasedcontentofintracellularglutathioneandactivityofglut