干细胞标志CD90与人肺腺癌A549细胞顺铂耐药之间的关系  

The relationship between tumor stem cells marker CD90 and resistance to cisplatin in human lung adenocarcinoma A549 cells

作　　者：况里杉 周向东 

机构地区：[1]第三军医大学第一附属医院呼吸内科,重庆400038

出　　处：《肿瘤》2013年第9期770-775,共6页Tumor

基　　金：国家重点基础研究发展计划(973计划)(编号:2010CB529402)

摘　　要：目的:观察干细胞标志CD90与人肺腺癌A549细胞顺铂(cisplatin,CDDP)耐药性的关系,从耐药角度探讨CD90是否是肺癌干细胞的标志。方法:应用成球实验、CCK-8(cell counting kit-8)和平板克隆形成实验检测A549和A549/CDDP细胞的自我更新能力,FCM法检测A549和A549/CDDP贴壁细胞及细胞球中CD90^+细胞的比例,细胞毒性实验检测CD90^+A549和CD90^-A549细胞对CDDP的耐药性。建立CD90^+A549和CD90^-A549细胞裸鼠移植瘤模型,检测体内CD90^+A549细胞对CDDP的耐药性。结果:与A549细胞比较,A549/CDDP细胞的成球能力、细胞增殖及克隆形成能力更强(P<0.01)。A549/CDDP贴壁细胞和细胞球中CD90^+细胞比例分别高于A549贴壁细胞和细胞球(P<0.05)。CDDP作用CD90^+A549细胞和CD90^-A549细胞的IC_(50)值分别为8.50和1.50μmol/L。经CDDP干预后,CD90^+A549细胞裸鼠移植瘤体积大于CD90^-A549细胞(P<0.05)。结论:A549/CDDP细胞具有较强的自我更新能力和较高比例的CD90^+亚群细胞,CD90^+A549细胞表现出较高的CDDP耐药性,提示CD90可能是肺癌的干细胞标志物。Objective： To observe the relationship between stem cells marker CD90 and CDDP （cisplatin） resistance in lung adenocarcinoma A549 cells, and to explore whether CD90 is a marker of lung cancer stem cells. Methods： Self-renewal abilities of A549 and A549/CDDP cells were detected by sphere-forming assay, CCK-8 （cell counting kit-8） and plate colony-formation assay, respectively. The proportions of CD90^＋ in A549 cells, A549/CDDP adherent cells and the spheres cells were measured by FCM （flow cytometry）. The CDDP resistance in CD90^＋A549 cells and CD90^-A549 cells was detected by cytotoxicity experiment. The CDDP resistance in CD90%＋A549 cells in vivo was detected by using transplanted tumor in nude mice. Results： As compared with A549 cells, the abilities of sphere formation, proliferation and colony-formation efficiency of A549/CDDP cells were increased （P〈0.01）. The proportions of CD90^＋ in A549/CDDP adherent cells and the sphere cells were higher than those in the corresponding A549 adherent cells and the sphere cells （P〈0.05）. IC50 （half inhibitory concentration） values of CDDP in CD90^＋A549 and CD90^-A549 cells were 8.50 and 1.50 μmol/L, respectively. The tumor xenograft volume of CD90^＋A549 cells was larger than that of CD90^-A549 cells after treatment with CDDP in nude mice （P〈0.05）. Conclusion： A549/CDDP cells show high self-renewal ability and CD90^＋ proportion. CD90^＋A549 cells have high ability of CDDP resistance. CD90 may be a marker of lung cancer stem cells.

关 键 词：肺肿瘤 肿瘤干细胞 顺铂 抗药性 肿瘤 CD90 A549细胞 

分 类 号：R734.2[医药卫生—肿瘤]