Assay Description
Genes such as TNF, BAD, CASP1 (ICE), BAX, BID, TP53, BIK, PYCARD (TMS1/ASC), FADD, BAD, BNIP1, CASP1 BNIP3L, TP53BP2, NOD1, TNFRSF10A, FASLG and TNFRSF25 (DR3) are examples of genes responsible for the induction of apoptosis. A number of genes (BCL10, BAG3, BCL2A1 (Bfl-1/A1), NOL3, BCL2L10, BFAR, BIRC6, BNIP1, BRAF, CD40LG (TNFSF5), IGF1R and TP73) have also been reported to be involved in the regulation of apoptosis as well. As apoptosis is a complex process, which is regulated by a large number of genes, a number of in vitro methods have been used to evaluate the effects of drugs on the expression of apoptosis pathways or regulator genes or proteins. For gene and protein expression studies real-time polymerase chain reaction (PCR), PCR arrays, microarrays, immunoprecipitation, western blotting and immunochemistry are the widely used in vitro techniques.
Assay Type Details
Apoptosis (programmed cell death) plays a vital role in embryonic development, homeostasis, functioning of immune system and wound repair. The ability to evade induction of apoptosis has been used by cancer cells to survive against host defense mechanisms. The molecular mechanisms involved in cancer cell apoptosis have been well documented and it involves certain biochemical events such as DNA fragmentation, chromatin condensation, cell organelle degradation and protein cleavage, etc. The extrinsic and intrinsic (mitochondrial) pathways are the two major pathways involved in apoptosis. With the available techniques and assays, a number of apoptosis inducing agents (natural compounds, synthetic compounds, nano-formulations, peptides and enzymes) in many cancer cells have been identified. Selection of an assay for apoptosis detection is based on factors such as apoptotic pathway, nature of drug, cell type being used and the method of analysis.
