中药土贝母诱导人乳腺癌细胞凋亡的实时成像  

Real-time imaging of induction of apoptosis of human breast cancer cell by traditional Chinese medicine Bolbostemma Paniculatun

作　　者：胡明昕[1] 安超[2] 李泉旺[2] Robert M.Hoffman 赵明 杨萌 傅延龄[5] 倪胜楼[5] 胡凯文[2] 

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学东方医院肿瘤科,北京100078 [3]美国安泰康生物技术有限公司,美国圣地亚哥92111 [4]美国加州大学外科系,美国圣地亚哥92103 [5]北京中医药大学国际学院,北京100029

出　　处：《中华中医药杂志》2012年第7期1810-1814,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国际合作项目课题(No.2009DFA31490)~~

摘　　要：目的:采用先进的荧光实时显像技术对中药土贝母植物块茎提取物抗乳腺癌作用进行评价。方法:MDA-MB-231人乳腺癌细胞被设计成为细胞质红色荧光蛋白(RFP)基因表达而细胞核组蛋白H2B绿色荧光蛋白(GFP)基因表达的双色荧光细胞,并分别在2D平面塑料平皿及3D立体明胶海绵块两种不同的培养环境中培养,经土贝母鲜品二氯甲烷提取物(DEFT)干预后,荧光共聚焦显微镜下观察细胞形态及核碎片形成的情况;DNA电泳方法进一步检测细胞凋亡。结果:土贝母鲜品二氯甲烷提取物作用24h后荧光显微镜下即观察到MDA-MB-231细胞出现凋亡的形态学改变;DNA电泳在药物干预48h后检测到凋亡的DNA碎片梯带,72h后更加明显;3D明胶海绵培养的MDA-MB-231细胞经实验药物作用后其形态学改变的效果不如2D环境下明显。结论:土贝母鲜品二氯甲烷提取成分具有抗乳腺癌治疗的潜能。Objective： In the present study,the efficacy on human breast cancer cell line of TCM Bolbostemma paniculatum,extracted from tubeus of the plant Bolbostemma paniculatum,was evaluated using advanced real-time fluorescence imaging system.Methods： The MDA-MB-231 human breast cancer cell line was engineered to express red fluorescent protein（RFP） in the cytoplasm and histone H2B-GFP in the nucleus which allows real time imaging of nuclear-cytoplasmic dynamics.The MDA-MB-231（RFP-GFP） cells were cultured either in 2-Dimensions（2D）on plastic or in 3-Dimensions（3D） on gelfoam.The cells were treated with dichloromethane extract of fresh tubeimu（DEFT）.Apoptosis was further monitored by DNA fragmentation determined by gel electrophoresis.Results： Bolbostemma paniculatum induced apoptosis of MDA-MB-231 cells was observed by fluorescence microscopy as early as 24 hours of treatment in vitro.By 48 hours treatement,DNA fragmentation could be observed,and the frequency of apoptosis increased after 72 hours treatment.The apoptosis morphology change of MDA-MB-231 cells in 3D culture condition which were treated by DEFT was to a lesser extent than in 2D culture.Conclution： The results of the present study indicate the potential of Bolbostemma paniculatum for breast cancer therapy.

关 键 词：土贝母鲜品二氯甲烷提取物 双色荧光蛋白 双色人乳腺癌细胞MDA-MB-231 实时荧光显微成像 明胶海绵3D培养 DNA电泳 凋亡 

分 类 号：R273[医药卫生—中西医结合] R737.9[医药卫生—中医肿瘤科]