新型光敏剂叶绿酸f对人膀胱癌T24细胞的光动力杀伤作用

将自主研发的新型光敏剂叶绿酸f与T24细胞共同孵育后,以650nm激光照射,通过MTT法检测光动力处理后T24细胞的生长抑制率,流式细胞术检测凋亡情况,同时利用激光共聚焦显微镜488nm和405nm双波长激发,应用线粒体探针,进行叶绿酸f的亚细胞定位。结果显示叶绿酸f为浓度2.5,5,10μg/mL对T24细胞生长抑制率在光能量密度4J/cm2下为17.68%,49.35%和84.42%,在1J/cm2时为4.34%,37.42%和78.38%;流式细胞术显示T24细胞光动力处理后,凋亡率为(45.23±1.2)%,显著高于对照组;激光共聚焦显微镜显示504nm激发叶绿酸f发出的红光与488nm激发线粒体探针产生的绿光分布基本一致。实验证明叶绿酸f对T24细胞杀伤效果明显,主要是通过作用于线粒体,诱导细胞凋亡来完成的。

Photodynamic Effect of Novel Photosensitizer Chlorophyllin f on Bladder Cancer T24 Cells

T24 cells are incubated with a novel photosensitizer chlorophyllin f, and then irradiated by 650 nm laser light. MTT assay is used to detect the growth inhibition rate of T24 cells. The apoptosis of T24 cells is observed by flow cytometry. The Mito Tracker Green probe is used to label mitochondrion, and the confocal laser scanning microscopy excited by 488 nm and 405 nm dual channels is applied to reveal intracellular localization. With 4 J/cm2 of optical energy density, the growth inhibition rates of 2.5, 5 and 10 μg/mL chlorophyllin f are 17.68%, 49.35% and 84.42%, while with 1 J/cm2 optical energy density, the growth inhibition rates are 4.34%, 37.42% and 78.38% respectively. The apoptosis rate of treatment group is (45.23±1.2)%, significantly higher than control groups. The confocal laser scanning microscopy reveals that, the red fluorescence emitted by chlorophyllin f and green fluorescence emitted by Mito Tracker Green probe distributes almost the same. Therefore, the photodynamic effect of chlorophyllin f on T24 cells is significant. Chlorophyllin f mainly locates and functions at mitochondria, and induces apoptosis, which may be one of the mechanisms against T24 cells.