A549肺癌细胞球抗凋亡能力及耐药蛋白表达分析  

Anti-apoptosis competence and drug-resistance protein expression of tumor spheres from lung cancer cell line A549

作　　者：孙芬芬[1] 贺斌峰[2] 余时沧[3] 黄学德[1] 叶小群[1] 

机构地区：[1]南昌大学第二附属医院呼吸内科,南昌330006 [2]第三军医大学新桥医院全军呼吸病研究所,重庆400037 [3]第三军医大学西南医院病理科,重庆400037

出　　处：《肿瘤》2011年第2期111-116,共6页Tumor

基　　金：江西省科技厅面上基金资助项目(编号:2009BSB11107)

摘　　要：目的:探讨肺腺癌A549细胞来源的肿瘤细胞球对化疗药物的抗凋亡能力及耐药蛋白谷胱苷肽-S-转移酶-π(glutathione-S-transferase-π,GST-π)的表达情况。方法:利用无血清培养法获得A549细胞球,将A549细胞球细胞和原代A549细胞株经顺铂处理后,采用Annexin Ⅴ-FITC+碘化丙啶(propidium iodie,PI)双染和JC-1染色的免疫荧光法检测2组细胞的凋亡情况。通过Western印迹法及荧光免疫组织化学法测定2组细胞及2组细胞成瘤组织中GST-π蛋白的表达水平。结果:经免疫荧光检测发现,A549细胞的凋亡率明显高于A549细胞球细胞。而A549细胞球细胞及其成瘤组织中GST-π蛋白的表达水平较A549细胞及其成瘤组织的高。结论:采用无血清培养法培养获得A549细胞球细胞的耐药性比A549细胞株高,这可能与细胞球细胞中耐药相关蛋白的高表达有关。Objective：To investigate the anti-apoptosis competence and the expression of glutathione-S-transferase-π（GST-π） of tumor spheres from lung cancer cell line A549.Methods：The tumor spheres were achieved from lung cancer cell line A549 cultured in serum-free medium.The apoptosis rates of both tumor spheres and primary A549 cells after treatment with cisplatin were detected by immunofluorescence assay with AnnexinⅤ-FITC＋PI and JC-1 stainings,and the expressions of GST-π protein were also examined in the tumor spheres and the primary A549 cells as well as their grafted tumors in nude mice by Western blotting and fluorescence immunohistochemistry,respectively.Results：Immunofluorescence assay showed that the apoptosis rate was significantly higher in tumor spheres than that in primary A549 cells.Compared with the primary A549 cells and their corresponding grafted tumors,the expression levels of GST-π protein were significantly higher in the tumor spheres and their corresponding grafted tumors.Conclusion：Tumor spheres from lung cancer cell line A549 have a strong capability of drug-resistance,which is probably associated with the up-regulated expressions of drug-resistance proteins in these tumor spheres.

关 键 词：肺肿瘤 球形体 细胞 谷胱苷肽-S-转移酶π 细胞凋亡 多药耐药相关蛋白质类 顺铂 

分 类 号：R734.2[医药卫生—肿瘤]