土木香内酯对RPMI-8226细胞的增殖抑制作用及其相关机制研究  

Effect of Alantolactone on Proliferation of RPMI-8226 Cells and Its Possible Mechnism

作　　者：姚瑶[1] 孙月月 夏丹丹[1] 牛铭山[1] 赵恺[1] 李振宇[1] 曾令宇[1] 徐开林[1] 

机构地区：[1]徐州医学院附属医院血液科,江苏徐州221006

出　　处：《中国实验血液学杂志》2015年第5期1336-1340,共5页Journal of Experimental Hematology

基　　金：国家自然科学青年基金(81302034)

摘　　要：目的:探讨土木香内酯(alantolactone)对多发性骨髓瘤RPMI-8226细胞增殖、凋亡的影响,并分析其可能的作用机制。方法:使用浓度为1、2.5、5、7.5及10μmol/L的土木香内酯处理RPMI-8226细胞48 h,应用CCK-8检测细胞活力,并计算其半数抑制浓度(IC50)值;使用浓度为2.5,5及7.5μmol/L的土木香内酯处理RPMI-8226细胞48 h后,应用AnnexinV/PI双标记法分析细胞凋亡;Western blot方法检测呈切割的caspase-3及ERK信号通路的变化;裸鼠荷瘤并给予土木香内酯以进一步验证其对多发性骨髓瘤细胞的促凋亡作用。结果:土木香内酯可抑制RPMI-8226细胞活力,并呈现剂量依赖效应(r=-0.9784,P=0.0007);RPMI-8226细胞48 h的IC50为4.32±0.15μmol/L;流式细胞术分析凋亡结果显示,随着土木香内酯浓度的增高,细胞早期凋亡率显著增加(r=0.9601,P<0.05);Western blot结果显示,土木香内酯处理RPMI-8226细胞后,活化了caspase-3,降低ERK磷酸化水平;体内结果显示,给药组小鼠瘤体积明显小于对照组(P<0.05);免疫组织化学检测结果显示,与对照组相比,实验组瘤内细胞核增殖相关抗原Ki67及p-ERK的水平降低。结论:土木香内酯可有效地抑制多发性骨髓瘤RPMI-8226细胞增殖并促进其凋亡,其机制可能是通过抑制ERK信号通路的活性,抑制裸鼠皮下多发性骨髓瘤的生长。Objective: To investigate the effect of alantolactone on perliferation and apoptosis of multiple myeloma( MM) RPMI-8226 cells,and to explore its possible mechism in vitro and in vivo. Methods: The RPMI-8226 cells were treated with alantolactone( 1,2. 5,5,7. 5 and 10 μmol /L) for 48 h,cell viability was detected by CCK-8 assay and the value of IC50 was calculated; The RPMI-8226 cells were treated with alantolactone( 2. 5,5 and 7. 5 μmol /L) for 48 h,the apoptotic rate was detected by flow cytmetry with Annexin V /PI staining; the expression level of cleaved caspase-3 and phosphorylation of ERK were measured by Western blot; the nude mice was used to further confirm the proapoptotic effect of alantolactone on MM cells in vivo. Results: The alantolactone inhibited RPMI-8226 cell viability remarkably with a dose-dependent manner; the IC50 value of RPMI-8226 cells at 48 h was 4. 32 ± 0. 15 μmol /L; the apoptotic rate increased observably with a dose-dependent manner; the levels of cleaved-caspase-3 increased and the phosphorylation of ERK decreased significantly; as compared to control,the volum of tumor was much smaller,the expression levels of Ki67 and p-ERK decreased. Conclusion: The alantolactone can efficiently inhibit the proliferation and induce the apoptosis of multiple myeloma RPMI-8226 cells in vitro and in vivo through inhibiting the activation of ERK signal pathway.

关 键 词：土木香内酯 多发性骨髓瘤 RPMI-8226细胞 ERK通路 细胞凋亡 

分 类 号：R285[医药卫生—中药学]