九节龙-Ⅲ经BAD凋亡途径诱导胶质瘤U251细胞凋亡  

Ardipusilloside-Ⅲ induced apoptosis in glioblastoma U251 cells through BAD apoptotic pathway

作　　者：林洪[1] 章翔[1] 程光[1] 汤海峰[2] 章薇[1] 董文鹏[1] 甄海宁[1] 曹卫东[1] 

机构地区：[1]第四军医大学西京脑科医院神经外科,陕西西安710032 [2]第四军医大学西京医院药剂科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2008年第5期431-435,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30370512;30873402)

摘　　要：目的研究九节龙-Ⅲ诱导胶质瘤U251细胞凋亡及其内在机制。方法四甲基偶氨唑蓝(MTT)分析药物对U251细胞及胶质细胞增殖的影响;膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)染色、Hoechst33342染色和透射电镜检测细胞凋亡;Western blotting分析Bcl-xL/Bcl-2相关死亡启动子(BAD)蛋白表达和磷酸化水平。结果九节龙-Ⅲ以剂量和时间依赖方式抑制U251细胞活性(P<0.05,IC50=8.2μg/ml),诱导染色质浓聚边集、凋亡小体形成等凋亡改变,且浓度低于40μg/ml时不抑制胶质细胞活性(P>0.05)。Western blotting发现U251凋亡细胞内BAD表达明显增加、去磷酸化并被裂解。结论九节龙-Ⅲ经BAD去磷酸化和裂解的凋亡途径诱导了胶质瘤U251细胞的显著凋亡。Objective To investigate the induced apoptosis in the glioblastoma U251 cells by ardipusilloside-Ⅲ in vitro and the underlying mechanisms. Methods 3-（4, 5-dimethylthiazol-2-yl）-2, 5- diphenyhetrazolium bromide assay was used to detect the effect of ardipusilloside-m on the proliferation of U251 cells and the primary cultured gliocytes. Annexin Ⅴ-fluorescein isothiocyanate/propidium iodide staining method, Hoechst 33342 stain and the transmission electron microscope were used to observe the apeptosis of U251 cells. Western blot analysis was used to detect the levels of total Bcl-xL/Bcl-2-associated death promoter homolog （BAD） and the phospho-BAD. Results Ardipusilloside-Ⅲ suppressed the proliferation of U251 cells in a doseand time-dependent manner （P 〈 0. 05, IC50=8.2μ/ml ） and induced typical apoptotic changes such as chromatin condensation and apoptotic body formation, however, ardipusilloside-m at concentration below 40μg/ ml did not suppress the proliferation of the normal gliocytes （P 〉0. 05）. Western blot analysis demonstrated the increased expression, evident dephosphorylation and cleavage of BAD which generated the truncated small BAD in U251 cells. Conclusion Ardipusilloside-Ⅲ can induce significant apoptosis through BAD dephosphorylation and cleavage in human glioblastoma U251 cells.

关 键 词：九节龙-Ⅲ 恶性胶质瘤 凋亡 BAD 

分 类 号：R730.2[医药卫生—肿瘤] R739.4[医药卫生—临床医学]