小白菊内酯对多发性骨髓瘤细胞的生长抑制及其凋亡诱导作用  

Proliferation inhibiting and apoptosis inducing effects of parthenolide on human multiple myeloma cells

作　　者：陈智超[1] 李秋柏[1] 邵菁[1] 吕建[1] 游泳[1] 仲照东[1] 邹萍[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022

出　　处：《中华医学杂志》2006年第28期1993-1996,共4页National Medical Journal of China

摘　　要：目的探讨小白菊内酯(parthenohde,VTL)对多发性骨髓瘤(multiple myeloma,MM)细胞的体外作用及其机制。方法培养人MM细胞系PRMI8266,在不同浓度的PTL作用不同时间后,以四甲基偶氮唑盐比色法(MTT法)及台盼蓝拒染检测细胞增殖及活性,用AO／EB染色荧光显微镜及Wright-Giemsa染色观察细胞形态学改变,应用膜联蛋白V(Annexin V)和碘化丙啶(PI)染色流式细胞仪检测细胞凋亡率,酶底物法测定细胞半胱氨酸蛋白水解酶3(Caspase-3)活性。结果(1)1- 10μmol／L的PTL对MM细胞生长曲线和活性有明显抑制作用,并呈时间和剂量依赖性,5μmol／L作用48 h,可达到接近50％的活性抑制;(2)5μmol／L PTL作用48 h后可见到细胞出现明显形态学改变,细胞形态变小、胞质减少、胞体胞核固缩、出现凋亡小体;(3)2μmol／L、5μmol／L、10μmol／L的PTL作用48 h后MM细胞的凋亡率分别为17．1％±2．6％、33．6％±3．8％、40．9％±3．1％,与对照组5．6％±1．2％相比,差异有显著统计学意义(均P<0．01);(4)PTL作用48 h后,MM细胞caspase-3活性明显增强,且呈浓度依赖性(P<0．01)。结论。PTL能明显抑制MM细胞体外生长及活性,其作用机制为增强caspase-3活性而诱导MM细胞凋亡;PTL作为新的MM细胞凋亡诱导剂,其作用靶点及体内效应值得深入研究。Objective To investigate the effect of parthenolide （ PTL ） on human multiple myeloma （MM） cells in vitro and its mechanism. Methods Human MM ceils of the line PRMI8266 were cultured and treated with PTL of the concentrations of 1,2.5, 5, 7.5, and 10 p, moL/L for 24, 48, or 72 hours. MM ceils treated with DMSO were used as control group. The optical density was measured so as to draw a growth curve. The cell viability was detected hy MTT and trypan-blue exclusion. The apeptosis was detected by flow cytometry. AO/EB staining and Wright-Giemsa staining were used to observe the morphological changes of the cells by fluorescence microscope and light microscope respectively. The caspase-3 activity was evaluated by BD ApoAlertTM Caspase Colorimetric Assay Kit. Results PTL significantly inhibited the proliferation and viability of the MM ceils time and dose-dependently （ all P 〈 0.01 ）, and significantly induced the ceil apeptosis after 48 h in a dose-dependent manner（P 〈0.01 ）. The early cell apeptosis rates for PTL of the concentrations of 2, 5 and 10 μmol/L were 17. 1% ± 2.6%, 33.6% ± 3.8%, and 40. 9% ± 3. 1% respectively, all significantly higher than that of the control group （5.6% ± 1.2%, all P 〈0.01 ）. The MM ceils treated with PTL of the concentration of 5 μmol/L for 48 h showed typical ceil apoptotic features. The caspase-3 activity of the MM cells was enhanced significantly by PTL in a time and dose- dependent manner （ all P 〈0.01 ）. Conclusion This first report of anti-proliferation and apeptosis induction effects of PTL on MM ceils shows that able to significantly inhibit the proliferation and induce the apeptosis of MM ceils and enhance the caspase-3 activity, PTL may be a potentially useful drug for treatment of MM.

关 键 词：小白菊内酯 多发性骨髓瘤 生长抑制 脱噬作用 

分 类 号：R733.3[医药卫生—肿瘤]