title: Multiplex analysis of anti-apoptotic BCL2 family and caspase 3 activation by microbead arrays.
abstract: We have developed a multiplex assay to measure the expression of anti-apoptotic proteins and caspase 3 activation using the Luminex platform. In this report, we show three applications for this assay. First, we used this assay to identify biomarkers for BCL2 inhibitors to obtain a quantitative measure of expression of anti-apoptotic proteins (BCL2, BCLxL, and MCL1) in a panel of cell lines and correlated their response to BCL2/BCLxL inhibitor, ABT-263 (navitoclax). Second, we used this assay to monitor the change of MCL1 protein expression and induction of active caspase 3 after treatment with cyclin-dependent kinase inhibitor flavopiridol. Finally, we used this assay to screen for small molecules that decrease MCL1 protein and identified new combinations with ABT-263. This method provides a quick and convenient way to measure basal expression of the anti-apoptotic proteins and monitor expression change upon drug treatment. It is also applicable for high-throughput screening for compounds that decrease the expression of these anti-apoptotic proteins.
authors.affiliation: Research and Development, AbbVie , North Chicago, Illinois.
authors.forename: Lloyd T
authors.initials: LT
authors.lastname: Lam
authors.forename: Lisa
authors.initials: L
authors.lastname: Roberts-Rapp
mesh_terms: D017209:Apoptosis; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism* / Q000473:pathology*; D053148:Caspase 3 / Q000378:metabolism*; D045744:Cell Line, Tumor; D004789:Enzyme Activation; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008863:Microspheres; D025941:Protein Interaction Mapping / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D053148:Caspase 3