CDHS801光动力诱导膀胱癌细胞凋亡的机制探讨

Mechanism of bladder cancer T24 cell apoptosis induced by CDHS801 photodynamic therapy

目的探讨光敏剂CDHS801光动力诱导膀胱癌细胞凋亡的机制。方法将膀胱癌T24细胞分成A、B、C、D组。A组为空白对照组;B组与CDHS801避光孵育;C组仅予激光照射;D组与CDHS801共同孵育后接受激光照射。倒置相差显微镜和电镜下观察各组细胞凋亡情况,激光共聚焦显微镜检测JC-1荧光探针标记的线粒体膜电位,Western blot法检测细胞质中的细胞色素C。结果 D组细胞变小变圆,染色质浓缩,染色质凝集边集、有新月体形成;其余三组细胞正常。细胞JC-1探针检测显示D组发出绿色荧光,其余三组细胞发出橙红色荧光。Westernblot结果显示仅D组细胞质中检出了细胞色素C。结论 CDHS801光动力诱导膀胱癌T24细胞凋亡的机制是治疗后线粒体膜电位下降,线粒体通透性增加,细胞色素C由线粒体移位到胞质中,启动了线粒体途径的细胞快速凋亡。

Objective To investigate the mechanism of in bladder cancer T24 cell apoptesis induced by CDHS801 photodynamic therapy. Methods TPA cell were cultured and divided into A, B, C, D groups. Group A was control group. Cells in group B were cultured with CDHS801. Cells in group C were illuminated with laser. Cells in group B were cultured with 5 mg/ml CDHS801 for 6 h, and then illuminated with 20 mW/cm^2 650 nm laser for 5 minutes . The apoptesis was evaluated by inverted phase contrast microscope and transmission eleotron microscope, the mitochondfia △ψm change was investigated by the specific fluorescence probe JC-1 with confocal fluorescence microimaging system. The changes of cytestolic cytochrome C was determined by Western blotting. Results In group D, the size of cells shrinkaged, chromatin condensed and apoptotic body emerged. And they were normal in the other groups. The fluorescence of the probe JCI-1 in group D was green, the other control groups were orange. Cytochrome C were detected in the cytoplasma of the T24 cell in group D only. Conclusions The mechanism of bladder cancer T24 cell apoptesis induced by CDHS801 photedynamie therapy is that CDHS801-based PDT can induce apoptosis of bladder 124 ceils through decreasing the mitoehondrial transmembrahe potential and inducing the release of eyt oehrome C.