Fas／FasL信号通路在千金子甾醇诱导HL-60细胞凋亡中的作用与机制研究  

Roles and mechanisms of Fas/FasL in the apoptosis of HL-60 cells induced by euphorbiasteroid

作　　者：郭菲[1] 李霞[2] 张超[3] 任霞[2] 史美艳[2] 姜国胜[2] 

机构地区：[1]山东省医学科学院基础医学研究所济南大学山东省医学科学院医学与生命科学学院 山东中医药大学附属医院检验科,济南250062 [2]山东省医学科学院基础医学研究所山东省医药卫生肿瘤免疫与中药免疫重点实验室山东省罕少见病重点实验室卫生部生物技术药物重点实验室国家中医药管理局免疫药理(三级)实验室山东省免疫生物靶向药物工程技术中心 [3]山东中医药大学药学院

出　　处：《国际肿瘤学杂志》2014年第9期679-684,共6页Journal of International Oncology

基　　金：基金项目：国家“十二五”科技支撑计划项目（2013BA107802）;国家自然科学基金（30771103）;山东省中药现代化重大科技计划项目（2008GG2NS02023）;山东省科技攻关政策扶持项目（2011GGC03083）;山东省科技发展计划项目（ZR2011HL045）;山东省中医药科技发展计划项目（2011-234）;山东中医药大学“名科工程”青年骨干培养计划（ZYDXY1328）

摘　　要：目的：在明确千金子甾醇诱导HL-60白血病细胞发生凋亡的基础上，从Fas/FasL信号通路的角度探讨其诱导凋亡的分子机制。方法 HL-60细胞培养体系中分别以千金子甾醇终浓度2．5、10、40μg/ml作用24 h后CCK-8法检测千金子甾醇对HL-60细胞增殖的抑制作用，光学及荧光显微镜下观察细胞形态学变化，Annexin Ⅴ/PI 流式细胞术检测细胞凋亡，反转录-聚合酶链反应法检测Fas/FasL、caspase-8和caspase-3 mRNA转录水平，比色法检测caspase-8和caspase-3的相对活性。结果千金子甾醇可明显抑制HL-60细胞的增殖，增殖抑制率分别为（34．9±3．7）％、（54．6±5．2）％、（61．3±4．3）％，细胞呈典型凋亡形态学改变，细胞早期凋亡率分别为（23．4±3．1）％、（35．7±4．3）％、（53．2±3．9）％。Fas、FasL、caspase-8和caspase-3 mRNA转录水平明显上调（P＜0．01），caspase-8和caspase-3活性显著增高（P＜0．01）。结论千金子甾醇可以诱导HL-60细胞发生剂量依赖性细胞凋亡，其机制与上调Fas/FasL凋亡信号通路有关。Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were （34.9 ±3.7）%,（54.6 ±5.2）% and （61.3 ±4.3）%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were （23.4 ±3.1）%,（35.7 ±4.3）% and （53.2 ±3.9）% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner （P＆lt;0.01 ）.After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced （P＆lt;0.01 ）.Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.

关 键 词：HL-60细胞 细胞凋亡 千金子甾醇 

分 类 号：R285[医药卫生—中药学]