摘要
目的探讨索拉菲尼联合细胞因子活化的杀伤细胞(cytokine-induced killer,CIK)对肝癌细胞的体外杀伤效应。方法取健康人外周血单个核细胞,经IFN-γ、IL-1、IL-2和抗CD3单抗诱导CIK细胞成熟,流式细胞仪检测培养第1、7、14天的CIK细胞表型。Cell Counting Kit-8试剂盒(CCK-8)检测索拉菲尼、CIK细胞及两者联合对肝癌细胞BEL-7402的体外杀伤效应,并用Annexin V-FITC细胞凋亡检测试剂盒检测肝癌细胞的凋亡率。结果CIK细胞经多种细胞因子诱导培养后,CD3+CD56+细胞比例明显上升,14d时CD3+CD56+细胞比例达(46.83±2.53)%,与第1天比差异有显著性(P<0.01),细胞量扩增了近1000倍。索拉菲尼和CIK细胞在适当浓度及效靶比联合应用时表现出杀伤活性增强(P<0.01)。索拉菲尼联合CIK细胞诱导的肝癌细胞BEL-7402凋亡率达(70.56±2.58)%,明显高于索拉菲尼组(45.38±1.76)%和CIK组(52.18±8.63)%(P<0.05)。结论索拉菲尼与细胞因子活化杀伤细胞疗法联合应用能明显增强对肝癌细胞的体外杀伤效应。
[ Objective ] To investigate the inhibitory effects of sorafenib combined with cytokine induced-killer （CIK） ceils on the expansion of hepatocellular carcinoma cell line BEL-7402 in vitro. [Methods] CIK ceils were generated in vitro by stimulation of peripheral blood mononuclear cells subsets with interferon-gamma （IFN-7）,IL- 1, IL-2 and anti-CD3 monoclonal antibody. Phenotype analysis of CIK cells on the 1, 7, 14 days was performed with flow cytometer （FCM）. The specific cytotoxicities of CIK ceils against ceil line BEL-7402 was determined by cell counting Kit-8. The Annexin V-FITC Apoptosis Derection Kit was used to detect apoptosis. [ Results ] Amount of CD3^＋CD56^＋ ceils were increased from （2.66±0.42）% to （46.83±2.53）% with elevated absolute amount over 1000 times after 2 week culture. Combined treatment of the cells resulted in significantly higher killing rate and apoptsis rate than that in the other groups （P 〈0.05）. [ Conclusion ] Sorafenib combined with CIK showed growth inhibition and induction of apoptosis in BEL-7402 ceils efficiently.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2009年第19期2885-2888,共4页
China Journal of Modern Medicine
