9-顺式维甲酸协同γ-干扰素诱导神经母细胞瘤分化、凋亡及生长抑制的实验研究  

9-cis retinoic acid and gamma-interferon act in synergy in inducing differentiation,apoptosis and growth inhibition in human neuroblastoma cells in vitro

作　　者：孙福涛[1] 张文同[1] 王旭平[2] 

机构地区：[1]山东大学齐鲁医院小儿外科,济南250012 [2]教育部和卫生部心血管重构与功能研究重点实验室

出　　处：《中华小儿外科杂志》2007年第5期258-261,共4页Chinese Journal of Pediatric Surgery

摘　　要：目的观察9-顺式维甲酸(9-cis retinoic acid,9-cis RA)联合γ-干扰素(gamma-interfer- on,γ-IFN)在体外诱导神经母细胞瘤(neuroblastoma,NB)SK-N-SH细胞分化、凋亡过程中细胞形态、周期的变化,探讨其协同作用的效果及其机制。方法将细胞分为A(对照组)、B(9-cis RA用药组)、C(γ-IFN用药组)和D(9-cis RA和γ-IFN联合用药组)4组,在处理后适当的时间分别观察细胞形态学变化,进行分化神经节细胞计数,用Hoechst-PI荧光染色法观察细胞的凋亡和死亡,四甲基偶氮唑盐(MTT)法测定NB细胞抑制率,流式细胞仪(flow cytometry,FCM)检测细胞周期分布及其凋亡和死亡。结果联合用药组细胞形态学分化明显,神经节细胞分化率显著高于其他各组(P<0.01);MTT法测定显示联合用药能显著提高NB细胞抑制率并明显优于单独用药;Hoechst-PI荧光染色及FCM钙依赖性磷脂结合蛋白V和碘化丙啶(annexin V/PD染色法检测一致显示联合用药对于诱导NB细胞早期凋亡和坏死具有显著的协同作用;FCM细胞周期动力学分析发现联合用药组G0-G1期细胞比率增加,S期细胞比率减少。结论9-cis RA联合γ-IFN在体外对NB细胞的分化、凋亡和生长抑制能产生明显的协同作用,为其联合应用于临床提供理论依据和指导。Objective To investigate the effects of the combination of 9-cis retinoic acid （9 cis RA） and gamma-interferon （γ-IFN） therapy in human neuroblastoma （NB） SK-N-SH cell line in vitro. Methods The cells were divided into four groups and treated with 9-cis RA, γ-IFN alone or in combination. At the desired time following treatment, neuronal cell differentiation was counted by e valuating morphological change; cell apoptosis and death were assessed with Hoechst-PI fluorescent staining; the growth inhibition rate was measured through MTT assay, and the cell cycle and apoptosis was assayed with a Flow cytometry （FCM）. Results In the combined treatment group, the differentiation of cell morphology was striking. The rate of differentiation was higher than the other groups （P〈0. 01）. MTT assay indicated that anti proliferative effects were more evident when the two agents were used in combination when compared to the effects of the drugs administered alone. Hoechst-PI and Annexin V/PI staining confirmed the significant synergistic effects of the combination on inducing cell apoptosis and death. We found increased G0-G1 stage cells and decreased S stage cells in the combined treatment group. Conclusions Taken together, these data confirm the synergistic effects of the combination of 9-cis RA and 7-IFN on inducing differentiation, apoptosis and growth inhibition of human NB cells. It also provides a rational basis and guidance for establishing a combination therapeutic approach for clinical use in the treatment of NB.

关 键 词：神经母细胞瘤 维甲酸 Γ干扰素 细胞分化 细胞凋亡 

分 类 号：R739.41[医药卫生—肿瘤]