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P1012
Predictive Cytotoxicity Measurements by Monitoring Heat Shock Proteins & the Cell Cycle in High-content Screening Cell-Based Assays
Presenter Suk Hong, Thermo Fisher Scientific, USA
Additional Authors: Richik N. Ghosh
Heat shock proteins (HSPs) are vital to all organisms, where they play crucial roles in cellular homeostasis and cell survival in response to a variety of cell stresses. HSPs have multiple cellular functions in signal transduction, cell survival and cell death, and protein dynamics. There are five major HSP families small HSPs, Hsp60 or chaperonins, Hsp70, Hsp90, and Hsp110. We developed a series of Thermo Scientific Cellomics® HCS Reagent Kits for quantitative, cell-based, high-content screening (HCS) of different HSPs to evaluate cellular stress responses. These kits and assays are designed for fluorescence labeling, imaging and detection of different HSPs in the cell (1) Hsp27 and phospho-Hsp27, (2) Hsp60 and Hsp90beta, (3) Hsp70 and Hsp90alpha, and (4) FKBP52 (Hsp59). To demonstrate the performance of these reagent kits, we used two representative cancer cell types, A549 and U2OS, treated them with different known compounds, and then imaged and analyzed the cellular responses with the HCS assays. The kits enabled us to precisely determine and evaluate the cell stress caused by toxic compounds, and the data were validated with independent cytotoxicity measurement (e.g., BrdU cell proliferation, p38 MAPK and cell loss). Cell stress responses by HSPs always precede animal toxicity, which is determined by LD50. The data provide not only excellent cytotoxicity indication but also unexpected cellular response against several toxic drugs. In summary, the use of automated, cell-based, high-content assays to monitor key HSPs is a suitable and effective strategy for cellular stress analysis as it enables the evaluation and toxicity profiling of different compounds.