摘要
目的研究多种细胞因子诱导的杀伤细胞(cytokine-induced killer cells,CIK)对A549肺癌细胞株诱导凋亡作用,并探讨其作用机制。方法应用末端脱氧核苷酸转移酶﹙TdT﹚介导的脱氧核苷酸缺口末端标志(TdT-mediated dUTP nick end labeling,TUNEL)法检测CIK细胞诱导凋亡的情况;通过免疫组织化学染色法检测A549细胞中凋亡相关基因p53、Fas、caspase-3,caspase-8,caspase-9、survivin蛋白,细胞增殖相关蛋白Ki-67的阳性表达率。结果 (1)TUNEL法检测结果:效靶细胞混合培养后,起初随时间延长凋亡细胞逐渐增多,5-14小时凋亡率明显上升,14-24小时凋亡率下降;(2)免疫组织化学染色结果表明,CIK实验组p53、Ki-67、survivin蛋白随作用时间延长而均下降,Fas、caspase-3、caspase-8、caspase-9蛋白表达上调,与对照组比较差异均有统计学意义(P<0.01)。结论 (1)CIK细胞对A549肺癌细胞具有抗增殖及诱导凋亡作用;(2)CIK细胞对A549肺癌细胞的抗增殖作用机制可能与下调Ki-67蛋白的表达,使癌细胞处于静止期有关;(3)CIK细胞对A549肺癌细胞的诱导凋亡作用机制可能与下调p53、survivin蛋白的表达,上调Fas、caspase-3、caspase-8、caspase-9蛋白的表达有关,经由细胞凋亡的死亡受体和线粒体通路完成凋亡的启动和执行;(4)CIK细胞是一种新型高效具有较强杀伤体外肺癌细胞的免疫活性细胞,有可能用于临床上晚期肺癌的过继性免疫治疗。
Objective To investigate the effect of cytokine-induced killer cells（CIKs） on human lung cancer A549 cells and its mechanism. Methods Apoptosis of A549 cells was detected by TUNEL method.The expressions of p53,Fas,caspase-3,caspase-8,caspase-9,survivin and Ki-67 proteins were determined by immunocytochemistry（ICC）. Results TUNEL showed that the apoptosis rate of A549 cells increased in 5-14 h and declined in 14-24 h after CIK treatment,which was significantly different from that of the control group（P0.01）.Immunocytochemistry showed that expressions of p53,Ki-67,and survivin proteins in A549 cells declined and Fas,caspase-3,caspase-8 and caspase-9 proteins increased with the time of CIK treatment,which was significantly different from that of the control group（P0.01）. Conclusion CIK can induce apoptosis of lung cancer A549 cells,which may be associated with the down-regulation of Ki-67,p53 and survivin proteins and up-regulation of Fas,caspase-3,caspase-8,and caspase-9 proteins.
出处
《实用肿瘤杂志》
CAS
北大核心
2011年第4期356-359,共4页
Journal of Practical Oncology
