Evaluating apoptosis in tumor cells infected with adenovirus expressing p53 tumor suppressor gene.
This chapter describes several methods for recognizing apoptosis in tumor cells following infection with a replication-deficient adenovirus expressing the tumor suppressor gene p53. We include cytotoxicity assays and assays of apoptosis, including DNA-nucleosomal DNA fragmentation (DNA laddering), TUNEL, DAPI staining, analysis of the sub-G1 (subdiploid) population, and degradation of poly(ADP-ribose) polymerase (as assayed by Western blot). Although this is not a comprehensive list of protocols to evaluate apoptosis, we believe that these will cover the majority of conditions of apoptosis that may arise. The chapter also describes the characteristics of each technique, including the advantages and disadvantages of each method.
Perron
Elizabeth
E
Laboratory of Gene Therapy, ENH-Research Institute and Department of Medicine, Evanston Hospital, Evanston, IL, USA.
Zafar
M Behzad
MB
Pister
Amanda
A
Wang
Zhen-Guo
ZG
Tian
Jun
J
Seth
Prem
P
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000502:physiology*; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D004279:DNA, Viral; D016158:Genes, p53; D006801:Humans; D020287:In Situ Nick-End Labeling; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D011065:Poly(ADP-ribose) Polymerases