Lenardo Michael J, McPhee Christina K, Yu Li
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Methods Enzymol. 2009;453:17-31. doi: 10.1016/S0076-6879(08)04002-0.
In this chapter we discuss methods to study autophagic cell death. A large body of evidence demonstrates that autophagy is a cell survival mechanism in response to starvation. The role of autophagy in cell death, however, has long been controversial. Recently, molecular approaches have provided direct evidence that autophagy contributes to cell death in certain contexts. We begin this chapter by outlining methods to quantify cell death, for example by assaying for cell viability. Next, we discuss methods to measure processes involved in cell death, such as caspase activation and autophagy. Finally, we discuss methods to genetically or chemically perturb autophagy to test whether autophagy is required for cell death. Together, these approaches provide a guide to investigate the relationship between autophagy and cell death.
在本章中,我们将讨论研究自噬性细胞死亡的方法。大量证据表明,自噬是细胞对饥饿作出反应的一种生存机制。然而,自噬在细胞死亡中的作用长期以来一直存在争议。最近,分子生物学方法提供了直接证据,表明在某些情况下自噬会导致细胞死亡。本章开篇将概述量化细胞死亡的方法,例如通过检测细胞活力。接下来,我们将讨论测量细胞死亡相关过程的方法,如半胱天冬酶激活和自噬。最后,我们将讨论通过基因或化学手段干扰自噬,以测试细胞死亡是否需要自噬。这些方法共同为研究自噬与细胞死亡之间的关系提供了指导。