全合成Jaspine B诱导人乳腺癌MDA-MB-231细胞凋亡及DNA损伤  

Apoptosis and DNA damage in MDA-MB-231 cells induced by total synthetic Jaspine B

作　　者：武建国[1] 董晓敏[1] 孟书聪[1] 丁宁[1] 霰海萍[1] 肖军军[1] 杜宇国[2] 

机构地区：[1]北京大学医学部细胞生物学系,北京100083 [2]中国科学院生态环境研究中心环境化学与生态毒理学国家重点实验室,北京100085

出　　处：《中国药理学通报》2008年第7期865-870,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30330690);教育部教育振兴行动计划专项资助项目(No985-2-016-24)

摘　　要：目的研究全合成Jaspine B对人乳腺癌MDA-MB-231细胞的增殖抑制及凋亡诱导作用,并探讨其对细胞DNA的损伤。方法酸性磷酸酶法(APA)检测细胞增殖,荧光显微镜、透射电镜(TEM)观察细胞形态结构,DNA琼脂糖凝胶电泳检测DNA片段化,流式细胞技术检测凋亡率及线粒体膜电位(ΔΨm),Western blot检测凋亡相关蛋白表达,单细胞凝胶电泳(SCGE)检测DNA损伤。结果Jaspine B抑制细胞增殖,24、48、72h IC50分别为2.61、1.07、0.77μmol·L-1,其作用在一定范围内呈浓度和时间依赖性;细胞发生凋亡形态改变,生化特征上出现DNA梯状改变;1.50μmol·L-1及3.00μmol·L-1Jaspine B作用24h,早期凋亡细胞率分别为7.60%及15.48%,相应浓度作用48h,早期凋亡细胞率分别增至21.48%及23.60%;Jaspine B可引起ΔΨm明显下降、Cyt C水平增加,促使Caspase-3酶原剪切活化;Jaspine B作用24h,细胞出现明显DNA损伤。结论全合成Jaspine B对人乳腺癌MDA-MB-231细胞抑制增殖、诱导凋亡并诱发DNA损伤,凋亡经由依赖胱天蛋白酶的内在途径发生。Aim To investigate the proliferation inhibiting, apoptosis inducing and DNA damaging effects on human breast cancer cell line MDA-MB-231 by total synthetic Jaspine B. Methods After MDA-MB-231 cells were incubated with Jaspine B, cell viability was evaluated by acid phosphatase assay （APA） ; fluorescent microscope and transmission electron microscope （TEM） were used to observe morphological changes; DNA agarose gel electrophoresis was used to detect the DNA fragmentation;flow cytometry was used to determine the apoptotic rate and mitochondrial membrane potential（△ψm） ;apoptosis related protein expression was analyzed by Western blot;single cell gel electrophoresis （SCGE） was employed to detect the DNA damage effect. Results Jaspine B inhibited MDA-MB- 231 cells proliferation significantly, somewhat in a dose-and-time dependent manner; IC50 at 24 h,48 h and 72 h were 2.61,1.07 and 0. 77 μmol·L^-1 respectively. Morphological changes and biochemical characteristic, DNA laddering of apoptosis were visual- ized. 1.50 μmol·L^-l and 3.00 μmol·L^-1 Jaspine B treated MDA-MB-231 cells for 24 h, early apoptosis rate were 7.60% and 15.48%, and when treatment time doubled to 48 h, early apoptosis rate increased to 21.48% and 23.60% respectively. Jaspine B caused △ψm disruption, enhanced Cyt C expression level and activated Caspase-3 cleaving to active forms. Exposed to Jaspine B for 24 h, DNA damage had been detected in MDA-MB-231 cells obviously. Conclusions Total synthetic Jaspine B inhibited proliferation and induced apoptosis and DNA damage in MDA-MB-231 cells;apoptosis was achieved through the Caspase-dependent intrinsic pathway.

关 键 词：Jaspine B MDA-MB-231细胞 凋亡 内在途径 DNA损伤 

分 类 号：R282.77[医药卫生—中药学] R286.0[医药卫生—中医学]