人肺腺癌A549细胞系干细胞样细胞的培养检测及其抗顺铂能力的研究  

ISOLATlON OF SPHERE CELLS IN HUMAN LUNG ADENOCARCINOMAS CELL LINE A549 AND ANALYSIS THE CISPLATIN RESISTANCE

作　　者：马晓雯[1] 刘彤[1] 孙力超[1] 刘军[1] 杨治华[1] 冉宇靓[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤研究所分子肿瘤学国家重点实验室哺乳动物细胞高效表达国家工程实验室抗体工程药物与肿瘤标志物中关村开放实验室,北京100021

出　　处：《广西医科大学学报》2011年第2期187-191,共5页Journal of Guangxi Medical University

基　　金：国家重点基础研究发展计划(973)项目(No.2009CB521804);国家科技重大专项课题(No.2009ZX09401-005和No.2009ZX09103-713)

摘　　要：目的:从人肺腺癌A549细胞系中培养获得含干细胞样细胞的球细胞(Sphere细胞),检测相关标志物表达及其对顺铂的耐药性。方法:采用含特定生长因子的无血清培养基诱导A549细胞的Sphere细胞形成,用亲脂荧光染料标记技术和免疫荧光法检测Sphere细胞中干细胞的存在,用流式细胞术检测干性标志物CD133,OCT4,CD56,CD117在Sphere细胞中的表达,用CCK8法检测顺铂处理后A549细胞及Sphere细胞的耐药性,并计算IC50。结果:A549细胞的Sphere细胞中含不对称分裂的干性标志物阳性细胞,且多种干性标志物表达比例升高,对顺铂耐药性也显著提高(P<0.05)。结论:人肺腺癌细胞系A549细胞在特定培养条件下可形成富集了干细胞样细胞的Sphere细胞,并对顺铂的抵抗能力增强。Objective：To induce the Sphere cells formation from human lung adenocareinoma cell line A549,and test the resistant of cisplatin.Methods：The Sphere cells were induced by specific growth factors with serum-free media,and tested the existence of stem cells with lipophilic fluorescent dyes.The stem cell related markers CD133,OCT4,CD56,CD117 were measured by FACS and Sphere cells-immunofluoreseent in the Sphere cells,and used CCK8 assay A549 Sphere cells after cisplatin treatment growth,and calculate the IC50.Results：A549 Sphere cells expressed the stem cell markers increased resistance to cisplatin is higher than the parent cells in significantly increased（P0.05）.Conclusion：The human lung adenocarcinoma cell line A549 in specific serum-free culture conditions can rich the stem cell-like cells,and its resistance to cisplatin was significantly increased.

关 键 词：人肺腺癌细胞 A549细胞 球细胞 顺铂耐药性 

分 类 号：R734.2[医药卫生—肿瘤]