PsD007诱导光动力疗法对人喉癌细胞的杀伤作用  

Killing Effect of Photodynamic Therapy Induced by PsD007 on Human Epidermoid Carcinoma in Vitro

作　　者：陈洪丽[1] 阴慧娟[2] 徐增瑞[3] 王明辉[3] 郑立卿[2] 李迎新[1,2] 

机构地区：[1]天津医科大学生物医学工程学院,天津300070 [2]中国医学科学院北京协和医学院生物医学工程研究所,天津300192 [3]天津市第四中心医院耳鼻喉科,天津300140

出　　处：《中国激光》2013年第5期133-139,共7页Chinese Journal of Lasers

基　　金：国家自然科学基金(31271434)资助课题

摘　　要：研究癌光啉(PsD007)诱导的光动力疗法(PDT)对人喉癌细胞Hep-2的杀伤作用,初步探索其杀伤机制。用噻唑蓝(MTT)法测定在不同光敏剂浓度、光照剂量以及抗氧化剂作用下的PDT杀伤效果;用共聚焦显微镜观察光敏剂的亚细胞定位;用Annexin V/PI双染法检测凋亡率;用DCFH-DA探针检测活性氧(ROS)产量。MTT结果显示PDT对Hep-2的杀伤效果可高达85.4%,光敏剂浓度和能量密度都与细胞死亡率呈正相关;三种抗氧化剂均能不同程度拮抗PDT对细胞的杀伤效应,以叠氮钠效果最好;光敏剂主要定位在线粒体;PDT 2h后ROS产量是对照组的2.71倍;PDT 4h后凋亡率可达49.5%(能量密度为1.2J/cm2)和70.2%(能量密度为4.8J/cm2)。PsD007-PDT对Hep-2细胞有明显的杀伤作用,并可诱导细胞凋亡,线粒体可能是其起始靶点。We investigate the killing effect of photodynamic therapy （PDT） induced by photocarcinorin （PsD007） on human epidermoid carcinoma Hep-2 in vitro, and explore the preliminary killing mechanism. Methyl thiazolyl tetrazolium （MTT） colorimetric assay is applied to measure the relative survival rate of PDT with different photosensitizer concentrations, light doses and antioxidants. Confocal microscope is used to observe the subcellular localization of PsD007. Annexin V/PI double staining is used to detect the apoptosis rate. DCFH-DA probe is used the to detect reactive oxygen species （ROS） production. The fatality rate of PDT on Hep-2 can be as high as 85.4%, and is positively correlated with photosensitizer concentration and energy density. Three kinds of antioxidants can all antagonize the damage effect induced by PDT in different degree, while sodium azide has the best effect. The fluorescence images of PsD007 subcellular localization demonstrate that PsD007 diffuses into the mitochondria. Two hours after PDT, the ROS production is as 2.71 times as control group. The apoptosis rate is up to 49.5 % （energy density is 1. 2 J/cm2 ） and 70. 2% （energy density is 4. 8 J/cm2 ） four hours after PDT. PsD007-PDT couldsignificantly kill Hep-2 cells and induce apoptosis that mitochondria may be the initial target.

关 键 词：医用光学 光动力疗法 癌光啉 人喉癌细胞 活性氧 

分 类 号：R318.51[医药卫生—生物医学工程]