光动力学疗法诱导犬乳腺肿瘤细胞凋亡及细胞Caspase-3和Caspase-8活性的变化  

Influence of hotodynamic therapy on apoptosis of canine breast tumor cells and activity of Caspase-3 and Caspase-8 in the process of apoptosis

作　　者：金鹏[1] 马兴乾[1] 任晓丽[1] 张蓉蓉[1] 李冉[1] 董慧[1] 刘云[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2009年第9期693-697,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：本文研究了光动力学疗法(PDT)促进犬乳腺肿瘤细胞凋亡的变化。通过不同剂量的光敏剂(HMME)和不同剂量的氦氖激光照射,用MTT法测定细胞的抑制率,确定抑制细胞生长的光敏感剂和激光照射的最佳剂量。采用最佳剂量组合作用细胞,对照射后不同时间点的细胞通过荧光显微镜(AO/EB染色)及透射电镜观察细胞的形态学变化;采用DNA LADDER法检测细胞凋亡;采用分光光度法测量照射后不同时间点的细胞内Caspase-3,Caspase-8的变化。研究表明光动力学的最佳剂量组为光敏感剂为20μg/mL,激光功率为2.8J/cm2。光动力学疗法作用细胞后,在荧光显微镜下观察出典型的细胞形态,并且随着光照时间的加长凋亡率逐渐增加,在透射电镜下细胞呈典型的凋亡形态,并且随着光照时间的延长凋亡的程度逐渐增加;细胞内Caspase-3,Caspase-8活性随照射后时间的增加其活性逐渐增强。由此可见光动力学疗法可有效的诱导犬乳腺肿瘤细胞凋亡,Caspase-3参与了细胞的凋亡过程,在细胞凋亡过程中起到一定的作用。To investigate the influence of photodynamic therapy （PDT） on the apoptosis of canine breast tumor cells, and examine the activity of Caspase-3 and Caspase-8 in the process of apoptosis. Using different Hematoporphyrinmonomethyl ether （HMME） concentrations and helium-neon laser doses to treat canine breast tumor cells of determine the best conditions of exposure. The Exposed cells were observed under fluorescence microscope, and apoptotic morphology examined under transmission electron microscope; Cell apoptosis was detected by DNA LADDER and the active state of Caspase-3 and Caspase-8 detected by spectrophotometric. Results mindicated that the best condition of the HMME-PDT was 20 txg/mL HMME with a laser dose of 2.8 J/cm^2. After HMME-PDT exposure, the cells were in the typical apoptosis state and characteristic apoptotic morphology under fluorescence microscope and transmission electron microscope. The apoptosis rate increased gradually over longer exposure; The activity of Caspase-3 and Caspase-8 also increased with longer time of treatment HMME-PDT can effectively induce the apoptosis of vitro on canine breast tumor cells; Caspase-3 and Caspase-8 participate and play a certain role in the process of apoptosis

关 键 词：光动力学疗法 光敏剂 凋亡 

分 类 号：S852.33[农业科学—基础兽医学]