CAJ | 학술논문

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。
목적：재명학천금자치순유도HL-60백혈병세포발생조망적기출상，종Fas/FasL신호통로적각도탐토기유도조망적분자궤제。방법 HL-60세포배양체계중분별이천금자치순종농도2．5、10、40μg/ml작용24 h후CCK-8법검측천금자치순대HL-60세포증식적억제작용，광학급형광현미경하관찰세포형태학변화，Annexin Ⅴ/PI 류식세포술검측세포조망，반전록-취합매련반응법검측Fas/FasL、caspase-8화caspase-3 mRNA전록수평，비색법검측caspase-8화caspase-3적상대활성。결과천금자치순가명현억제HL-60세포적증식，증식억제솔분별위（34．9±3．7）％、（54．6±5．2）％、（61．3±4．3）％，세포정전형조망형태학개변，세포조기조망솔분별위（23．4±3．1）％、（35．7±4．3）％、（53．2±3．9）％。Fas、FasL、caspase-8화caspase-3 mRNA전록수평명현상조（P＜0．01），caspase-8화caspase-3활성현저증고（P＜0．01）。결론천금자치순가이유도HL-60세포발생제량의뢰성세포조망，기궤제여상조Fas/FasL조망신호통로유관。
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<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.