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目的研究新型光敏剂2-丁胺-2-去甲氧基竹红菌乙素(BAHB)的光动力作用诱导人胰腺癌细胞系(Capan-1)凋亡的机制。方法利用荧光显微镜、细胞凋亡DNA梯带、激光共聚焦扫描显微镜、Western blot分析等技术研究BAHB诱导Capan-1凋亡与线粒体损伤的关系。结果低浓度BAHB主要定位于线粒体,高浓度时定位于溶酶体。DNA琼脂糖电泳分析显示出特征性细胞凋亡的DNA梯带。激光共聚焦扫描显微镜观察到线粒体跨膜电位(ΔΨm)降低,定量分析结果表明,随着光照时间的缩短,细胞的相对荧光强度增强。用Western blot法可以检测到线粒体中细胞色素C的释放。结论低浓度BAHB的光动力作用诱导人胰腺癌细胞凋亡的机制与线粒体损伤有关。
Aim To study the mechanism of apoptosis of Capan-1 cells induced by photodynamic therapy of 2-butylamine-2-demethoxyhomocholine (BAHB). Methods The relationship between apoptosis of Capan-1 induced by BAHB and mitochondrial damage was studied by fluorescence microscopy, DNA ladder, confocal laser scanning microscopy and Western blot. Results Low concentration of BAHB mainly located in the mitochondria, high concentration of lysosomes. DNA ladder analysis of DNA ladder showing characteristic apoptosis. The decrease of mitochondrial transmembrane potential (ΔΨm) was observed by confocal laser scanning microscopy. Quantitative analysis showed that the relative fluorescence intensity of cells increased with the illumination time shortened. Cytochrome C release from mitochondria was detected by Western blot. Conclusion The mechanism of photodynamic therapy of low concentration BAHB inducing apoptosis of human pancreatic cancer cells is related to mitochondrial damage.