A Proteomic Dataset of HepG2 Cellular Responses to Cycloheximide-Induced Translational Arrest
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Abstract
Cycloheximide (CHX), a potent translational inhibitor, is widely used in cellular assays requiring translational blockade. Due to limited characterization of CHX-exposed cellular models, this study investigated the molecular and cellular effects of CHX in human hepatoma HepG2 cells. Cells were treated with various CHX concentrations for early (1 and 3 hours) and prolonged (18 and 24 hours) periods and evaluated for cellular responses. MTT assays showed no significant viability changes during early exposure, whereas prolonged treatment with 25 and 50 uM CHX reduced viability by approximately 30%. Sustained CHX exposure induced Sphase cell cycle arrest with minimal apoptosis (approximately 10%). LC-MS/MS analysis identified 19,482 differentially expressed proteins (DEPs), while PLS-DA revealed clear separation between untreated and CHX-treated HepG2 proteomes across time points. Volcano plot analysis identified the five most altered proteins at each time point, representing diverse cellular compartments and biological functions. STITCH network analysis further identified three interactome clusters associated with CHX responses: calcium-mediated survival signaling, xenobiotic detoxification, and ubiquitin-proteasome degradation. This characterized CHX-exposed HepG2 model provides a useful methodological reference for bioassays requiring translational stress.
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