摘要
目的:探讨小檗胺(berbamine,BBM)体外诱导人多发性骨髓瘤RPMI 8226细胞的凋亡及其机制。方法:采用MTT法测定不同浓度BBM作用后细胞增殖抑制率并求得IC50;DNA凝胶电泳、流式细胞术分析细胞凋亡;RT-PCR检测BBM作用前后细胞p53、p21和GADD45 mRNA的表达;Western blot检测BBM作用前后细胞p53、JNK、p-JNK及c-Jun蛋白表达。结果:BBM抑制RPMI 8226细胞增殖呈剂量依赖性(P<0.05),48 h的IC50值为3.83μg/ml;8μg/ml BBM作用RPMI 8226细胞24 h后DNA凝胶电泳可见典型梯形条带,流式细胞术检测细胞凋亡率由1.07%升高至24.84%;BBM作用后细胞p53、p21及GADD45γmRNA表达上调,同时伴有胞核内p53蛋白上调及p-JNK、c-Jun蛋白活化。结论:BBM能抑制RPMI 8226细胞的增殖,其作用可能通过活化GADD45/JNK信号通路诱导细胞凋亡。
Objective： To investigate the effect of berbamine （BBM） on multiple myeloma （MM） cell line RPMI 8226 and its mechanism. Methods： MTT bioassay was used to examine the effect of berbamine on cell growth and IC50 was calculated. Apoptosis was observed by flow cytometry （FCM） and DNA gelose electrophoresis, p53,p21,GADD45 mRNA were measured by RT-PCR. The alterations in p53,JNK,p-JNK and c-Jun proteins were detected by Western blot method. Results： The growth of RPMI 8226 cells was suppressed in a dose-dependent manner after treatment with BBM（P〈0. 05）,and its IC50 value was 3.83 μg/ml at 48 h. Both DNA ladder and FCM results showed that BBM induced apoptosis of RPMI 8226 cells with concomitant increase of activated p53,p21 and GADD457 mRNA. Aftertreatment with BBM at 8 μg/ml for 24 h,the percentage of apoptotic cells increased from 1.07% to 24.84%. p-JNK and c-Jun proteins were activated. Conclusion： BBM can inhibit the growth of RPMI 8226 cells,which is associated with activation of,GADD45/JNK signaling pathway and induction of cell apoptosis.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2009年第5期439-444,共6页
Journal of Zhejiang University(Medical Sciences)
