摘要
目的:研究天花粉多糖(polysaccharide of snakegourd root)对人乳腺癌MCF-7细胞生长的抑制作用、诱导凋亡的作用以及Caspase-3、Caspase-8活化对凋亡的影响。方法:采用噻唑蓝(MTT)法观察不同浓度天花粉多糖对人乳腺癌MCF-7细胞的生长抑制作用;DAPI染色后荧光显微镜进行细胞凋亡核形态学观察;通过DNA琼脂糖凝胶电泳进行DNA片段化分析,FITC-Annexin V/PI荧光标记流式细胞术检测细胞凋亡率,分光光度比色法测定天花粉多糖诱导人乳腺癌MCF-7细胞凋亡过程中Caspase-3、Caspase-8活性变化。结果:MTT结果显示,天花粉多糖对人乳腺癌MCF-7细胞有生长抑制作用。10.0μmol/L以上浓度的天花粉多糖作用2 d,DAPI染色可见核浓缩及边缘现象,DNA电泳出现特征性的凋亡条带。1.0、5.0、10.0和20.0μmol/L天花粉多糖处理的MCF-7细胞2 d,细胞凋亡率分别为(5.2±1.3)%、(13.1±4.7)%、(27.6±6.8)%和(43.8±9.8)%;10.0μmol/L天花粉多糖处理MCF-7细胞1、2、3 d,Caspase-3活性在2 d达最高(2.32±0.12)U/μg,而Caspase-8活性则在1d达最高(1.92±0.11)U/μg,两者与对照组相比均有统计学显著性差异(P<0.05)。结论:天花粉多糖对人乳腺癌MCF-7细胞有生长抑制和诱导凋亡作用,其诱导凋亡的机制可能与Caspase-3、Caspase-8活化有关。
Objective： To investigate the effects of snakegourd root polysaccharide on apoptosis of human breast cancer cells（MCF-7 cells）.Methods： Colorimetric MTT assay was used to measure the inhibition of snakegourd root polysaccharide on MCF-7 cells.The morphological changes of MCF-7 cells were observed by fluorescence microscope after DAPI staining and transmission electron microscope.The apoptosis of MCF-7 cells was examined by DNA agarose gel electrophoresis analysis of DNA fragmentation amd flow cytometry.The activity of Caspase-3 and Caspase-8 was detected by colorimetric assay.Results： Polysaccharide of snakegourd root significantly inhibited MCF-7 cells in a dose-and time-dependent manner.The nuclear condensation and marginalization were observed by DAPI staining and transmission electron microscope.The characteristic ladder of apoptosis in DNA electrophoresis was detected in MCF-7 cells treated with 10.0 μmol/L polysaccharide of snakegourd root at d 2.The activities of Caspase-3 and Caspase-8 were increased in a time-dependent manner.The rates of apoptosis in MCF-7 cells were（5.2±1.3）%,（13.1±4.7）%,（27.6±6.8）% and（43.8±9.8）% treated with 1.0,5.0,10.0 and 20.0 μmol/L snakegourd root polysaccharide at d 2,respectively.The maximal activities of intracellular Caspase-3 and Caspase-8 were（2.32±0.12）U/μg and（1.92±0.11）U/μg at d 2 and d 1,respectively when MCF-7 cells were treated with 10.0 μmol/L.Conclusion： The polysaccharide of snakegourd root can induce the apoptosis of MCF-7 cells,which is associated with the activation of intracellular Caspase-3 and Caspase-8.
出处
《浙江大学学报（医学版）》
CAS
CSCD
北大核心
2012年第5期527-534,共8页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省中医药科技计划项目(2008CA108)
