Dewaele Michael, Verfaillie Tom, Martinet Wim, Agostinis Patrizia
Department of Molecular and Cell Biology, Catholic University of Leuven, Leuven, Belgium.
Methods Mol Biol. 2010;635:7-33. doi: 10.1007/978-1-60761-697-9_2.
Photodynamic therapy (PDT) is an anticancer modality utilizing the generation of singlet oxygen and other reactive oxygen species through visible light irradiation of a photosensitive dye accumulated in the cancerous tissue. Upon exposure of cancer cells to the photodynamic stress, multiple signaling cascades are concomitantly activated and depending on the subcellular location of the generated ROS and the intensity of the oxidative damage, they dictate whether cells will cope with the stress and survive or succumb and die. Different methodologies have been developed to allow the discrimination of cell death subroutines at the morphological, ultrastructural, and biochemical levels and to scrutinize signaling cascades in response to PDT. Here we describe a selection of useful techniques to characterize apoptosis and autophagy and to monitor the activation status of the MAPK- and Akt-mTOR pathways after PDT.
光动力疗法(PDT)是一种抗癌治疗方式,它通过对积聚在癌组织中的光敏染料进行可见光照射,产生单线态氧和其他活性氧物质。当癌细胞暴露于光动力应激时,多个信号级联反应会同时被激活,并且根据所产生的活性氧的亚细胞定位以及氧化损伤的强度,这些信号级联反应决定细胞是应对应激并存活,还是屈服并死亡。已经开发出不同的方法,以在形态学、超微结构和生化水平上区分细胞死亡子程序,并仔细研究响应PDT的信号级联反应。在这里,我们描述了一系列有用的技术,用于表征细胞凋亡和自噬,并监测PDT后丝裂原活化蛋白激酶(MAPK)和蛋白激酶B(Akt)-哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的激活状态。