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1、人卵巢癌细胞系A2780中肿瘤干细胞的分离及鉴定谢伟1，龙海霞1，张恒1，向橦1，朱波1，谢荣凯2（400037重庆，第三军医大学新桥医院：全军肿瘤研究所1.；妇产科）[摘要]目的：从人卵巢癌细胞系A2780中分离并鉴定卵巢癌干细胞。方法：将A2780细胞置于含EFG、bFGF、BSA和人胰岛素的无血清培养基中培养，采用连续传代诱导培养肿瘤干细胞，对每代分离出来的肿瘤干细胞进行流式检测，并对第四代肿瘤干细胞进行免疫荧光检测和逆向分化实验。结果：从A2780细胞中成功分离出肿瘤干细胞，在上述无血清培养基中呈悬浮生长，具有很强的自我更新能力，流式检测显示随
2、着传代诱导，CD133阳性率依次升高，免疫荧光进一步证实了CD133是卵巢癌干细胞的表面标记物，诱导分化实验说明其可逆向分化成为A2780肿瘤细胞。结论：体外培养的卵巢癌细胞株A2780中存在卵巢癌干细胞，并能将其分离培养和诱导分化。[关键词]卵巢癌；卵巢癌干细胞；分离；鉴定[中图分类号]R737.31IsolationandIdentificationofOvarianCancerstemcellsfromOvarianCancercelllineA2780XieWei1,LongHaixia1,ZhangHeng1,XiangTong1,ZhuBo
3、1,XieRongkai2(CancerResearchCenter1,DepartmentofGynaecologyandObstetrics2,XinqiaoHospital,ThirdMilitaryMedicalUniversity,Chongqing400037,China)[Abstract]Objective:ToisolateandidentifyovariancancerstemcellsfromOvariancancercelllineA2780invitro.Methods:TheA2780wereculturedinserum-
4、freemediumwithEGF,bFGF,insulinandBSA,bycontinuousinducedtumorstemcellsineverypassage,andflowcytometrywereappliedtoidentifythecancerstemcellineveypassage,moreovertheimmunofluorescenceandreversedifferentiationexperimentswereconductedinthefourthpassageofcancerstemcells.Results:Theo
5、variancancerstemcellshavebeensuccessfullyisolatedfromovariancancercelllineA2780.Theyformedtypicalneurospheresinserum-freemedium,andhadstrongself-renewcapacity.TheresultsofflowcytometryillustratedthatCD133positiverateincreasedwiththepassageofcancerstemcell.Theimmunofluorescencefu
6、rtherconfirmedthatCD133+cellsarethesurfacemarkerofovariancancerstemcells,andtheycandifferentiateintoovariancancercellsinreversedifferentiationexperiments.Conclusion:TheovariancancerstemcellsexistinovariancancercelllineA2780,andtheycanbeisolated,culturedanddifferentiated.[keyword
7、s]ovariancancer;ovariancancerstemcell;isolation;identificationSupportedbytheNationalScienceFoundationofChina(30901592).Correspondingauthor:XieRongkai，E-mail:xrkai@163.com肿瘤干细胞是肿瘤组织中具有自我更新能力并能产生异质性肿瘤细胞的细胞，具有连续增殖和向不同表型分化的能力。在肿瘤组织中，肿瘤干细胞能够分化为新的肿瘤细胞，其不对称的分裂导致了肿瘤组织的异质性[1]。它对化疗药物及放射治疗
8、有很强的抵抗性，已成为肿瘤治疗的瓶颈。越来越多的研究表明，肿瘤干细胞可能作为肿瘤临床治疗的一个
