title: Cellular Apoptosis Assay of Breast Cancer.
abstract: Apoptosis is an energy-dependent enzymatic cell suicide process. It almost always involves the activation of caspases. In this chapter, we systemically introduce methodologies to assay caspases dependent biochemical and morphological changes in vitro breast cancer cell lines and in vivo breast cancer tissues. In addition, mitochondrial involvement is crucial to distinguish two different apoptotic pathways. Methodology to assay dissipation of mitochondrial transmembrane potential, an early event of mitochondrial involvement, is also included. Of note, since apoptotic features may not appear to the same extent depending on the context of cell types and the death-inducing insults, a common practice is to use more than one method to assess apoptosis, qualitatively and quantitatively.
authors.affiliation: Department of Chemical Biology, Rutgers University, 164 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Department of Chemical Biology, Rutgers University, 164 Frelinghuysen Road, Piscataway, NJ, 08854, USA. weixing.zong@pharmacy.rutgers.edu.
authors.forename: Wei-Xing
authors.initials: WX
authors.lastname: Zong
mesh_terms: D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D001943:Breast Neoplasms / Q000473:pathology*; D020169:Caspases / Q000378:metabolism; D053938:DNA Fragmentation; D004789:Enzyme Activation; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D059748:Proteolysis; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D017304:Annexin A5; D020169:Caspases
keywords: Annexin V/PI staining; Apoptosis assay; Breast cancer; Caspase activation; DNA laddering assay; In situ TUNEL staining for apoptotic cells; JC-1 staining