光敏剂HMME对人骨肉瘤细胞U-2OS的光动力作用及其作用机制的初步研究  

Photodynamic Effect of HMME on Cell Line U-2OS and its Mechanism Analysis

作　　者：王卓莹[1,2] 曾辉[1,2] 孙梦熊[1,2] 蔡郑东[1,2] 廖宇昕[1,2] 左冬青[1,2] 付东[1,2] 华莹奇[1,2] 

机构地区：[1]上海市第十人民医院骨科 [2]同济大学骨肿瘤研究所,上海200072

出　　处：《现代生物医学进展》2013年第14期2622-2624,2673,共4页Progress in Modern Biomedicine

基　　金：上海市科学技术委员会基金项目(11JC1410101)

摘　　要：目的:探讨光敏剂(HMME)介导的光动力疗法对人骨肉瘤细胞U-2OS的杀伤效应及机制研究。方法:使用不同浓度(0、10、20、30、40μg.ml-1)的光敏剂,采用不同光照能量(0、3、6、9 J.cm-2)照射人骨肉瘤细胞,与空白对照组、药物对照组(无光照但加光敏剂)和光照对照组(不加光敏剂但加光照)进行比较,MTT法检测细胞的存活率,选择半数有效量药物浓度和光照能量,作为实验组。以空白对照组为对照,采用Hoechst33342染色法,观察细胞凋亡情况。用western blot方法检测细胞凋亡蛋白caspase-7、caspase-9和PARP-1。结果:MTT结果显示,空白对照组、药物对照组和光照对照组对细胞存活率在96.7%和100%之间,药物的半数有效量为40.1μg.ml-(16 J.cm-2)和25.0μg.ml-(19 J.cm-2)。Hoechst33342染色法观察到实验组细胞明显凋亡。westen blotting检测结果,实验组与对照组相比,caspase-7、caspase-9和PARP-1表达明显增高。结论:HMME-PDT对人骨肉瘤细胞U-2OS有显著的杀伤效应,且呈光敏剂浓度和光照强度依赖性,其杀伤效应与caspase途径相关。Objective： To evaluate the effect of HMME-mediated photodynamic therapy on human osteosarcoma cell line U-2OS,and to investigate the therapy-related apoptosis mechanism of U-2OS cell in vitro.Methods： Different laser doses（0,3,6,9 J·cm-2） were applied to irradiate the human U-2OS cell after the osteosarcoma cell had been treated with different concentrations of HMME（0,10,20,30,40 μg·ml-1） for 4 hours.MTT was employed to detect the cell survival to yield the EC50 dosage in the study group,control group,drug control group and laser control group.The EC50 corresponding HMME dosage and laser dose were identified as the standard for the study group.The apoptotic morphology of the study group was observed under optical microscopy after processed with Hoechst33342 nuclear staining method,compared with a blank control group.Western blot was applied to analyzed the expression of caspase-7,caspase-9 and PARP-1 protein.Results： U-2OS cell survival rate detected MTT ranged from 96.7 % to 100 %,EC50 for HMME were 40.1 μg·ml-1（6 J·cm-2）and 25.0 μg·ml-1（9 J·cm-2）.Western blot of caspase-7,caspase-9 and PARP-1 protein indicated a significant difference between study group and the control group.Conclusion： Human osteosarcoma cell line U-2OS cell can be treated with HMME-mediated photodynamic therapy by inducing the activation of caspase pathway in vitro.

关 键 词：光动力疗法 光敏剂HMME 骨肉瘤 细胞凋亡 

分 类 号：R738[医药卫生—肿瘤] R730.5[医药卫生—临床医学]