国产新型光敏剂CDHS801光动力学杀伤膀胱癌机理的实验研究  

Effect of new photosensitizer CDHS801-mediated photodynamic therapy on bladder cancer: an experimental study

作　　者：郑景存[1] 赵鸿[1] 陈波[1] 吴忠[1] 姜昊文[1] 丁强[1] 张元芳[1] 

机构地区：[1]复旦大学华山医院泌尿外科复旦大学华山医院泌尿外科研究所,上海200040

出　　处：《中华医学杂志》2005年第25期1762-1765,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30170938);高等学校博士学科点专项科研基金资助项目(20010246054)

摘　　要：目的研究光敏剂CDHS801光动力学治疗膀胱癌机理。方法将膀胱癌T24细胞与CDHS801共同孵育,采用激光共聚焦显微镜为成像工具,应用线粒体荧光探针MitoTrackerREDCMXRose和MitoTrackerGreenFM进行CDHS801的亚细胞定位分析。MTT法检测光动力学治疗后存活率,激光共聚焦显微镜丫啶橙/溴化乙锭染色观察T24染色情况,电镜检测细胞超微结构的变化以及流式细胞仪判断细胞凋亡情况。结果激光共聚焦显微镜下,CDHS801和线粒体荧光探针发出的荧光都位于细胞浆内,在核周呈点状散在分布。PDT后实验组细胞存活率明显降低,细胞膜完整性丧失被染成黄色,流式细胞仪PI染色在subG1期出现了凋亡峰,透射电镜下细胞染色质于核膜下浓聚边集形成新月体,细胞发生凋亡,对照组则没有这些表现。结论CDHS801经膀胱癌T24细胞摄取后,分布在细胞浆内,主要在线粒体内,激光照射后膀胱癌细胞发生了凋亡。Objective To investigate the effects of dynamic photodynamic therapy (PDT) on bladder cancer. Methods Human bladder cancer cells of the line T24 were co-cultured with CDHS801, a photosensitizer, and MitoTracker RED CMXRose and MitoTracker GREEN FM, mitochondria specific fluorescence probe dyes. Laser scanning confocal fluorescence microimaging system was applied to collect the fluorescence of the photosensitizer and the probes. T24 cells were cultured and divided into 4 groups: Group 1 as blank control group, Group 2 undergoing laser irradiation, Group 3, added with CDHS801 for 6h, and Group 4 (PDT group), added with CDHS801 and undergoing laser irradiation. The survival of the cells was examined by MTT colorimetric assay. The morphological changes and apoptosis of the photo-activated T24 cells were investigated by transmission electron microscopy, confocal laser scan microscopy, and flow cytometry. Results The fluorescence of the photosensitizer and that of the probes were detected in the cytoplasm and in the peri-nuclear region, mainly in the mitochondria, of the T24 cells. 2. The inhibitory rates of PDT on T24 cells were 0%, 7.3%, 10.8%, and 71.4% in the control group, Group 1, Group 2, and Group 3 respectively. T24 cell photo-activated with CDHS801 showed cell size shrinkage, condensed chromatin and formation of apoptotic body. Flow cytometry showed that apoptosis was seen in 55.31% of the photo-activated cells and peaked in the sub-G1 phase. However, no such changes were seen in the control group. Conclusion CDHS801-based PDT can kill bladder cancer T24 cells. CDHS801 is localized in the cytoplasm and peri-nuclear region, mainly mitochondria, of tumor cell. CDHS801 based PDT maybe eliminate the T24 cell by the induction of apoptosis.

关 键 词：CDHS801 新型光敏剂 实验研究 机理 激光共聚焦显微镜 膀胱癌T24细胞 流式细胞仪 杀伤 国产 光动力学治疗 MTT法检测 细胞超微结构 荧光探针 Green 细胞存活率 细胞染色质 癌细胞发生 成像工具 定位分析 染色观察 溴化乙锭 

分 类 号：R737.14[医药卫生—肿瘤] R739.7[医药卫生—临床医学]