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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 study the mechanism of photosensitizer CDHS801 photodynamic therapy for bladder cancer. Methods T24 cells of bladder cancer were co-incubated with CDHS801. The laser scanning confocal microscope (LSCM) was used as the imaging tool. The subcellular localization of CDHS801 was analyzed by MitoTracker REDCMXRose and MitoTracker Green FM. The survival rate after photodynamic therapy was measured by MTT method. T24 staining was observed by laser scanning confocal microscopy with acridine orange / ethidium bromide staining. The ultrastructural changes of the cells were observed by electron microscopy and apoptosis was evaluated by flow cytometry. Results Under laser scanning confocal microscopy, the fluorescence of CDHS801 and mitochondrial fluorescence probe were located in the cytoplasm, scattered in spots around the nucleus. After PDT, the survival rate of the experimental group was significantly decreased, the loss of cell membrane integrity was stained yellow, flow cytometry PI staining in the subG1 phase apoptosis peak, under the transmission electron microscope chromatin in the nuclear membrane under the concentration of the formation of a new set On the other hand, cell apoptosis occurred in the control group but not in the control group. Conclusion CDHS801 is expressed in the cytoplasm of bladder cancer T24 cells mainly in the mitochondria, and apoptosis of bladder cancer cells occurs after laser irradiation.