Gene expression profiles for genetically engineered human hepatoma hi-Hep cells with or without heat treatment, cultured as monolayer or spheroids using serum-containing or serum-free functional media

We developed a heat-inducible hepatic cell line (hi-Hep) in which BRCA1-associated protein-1 (BAP1) was introduced under the control of a TRE/PCMVmin promoter, in addition to the existing eight liver-enriched transcription factor (LETF) genes (HNF-1α, HNF-1β, HNF-3β, HNF-4α, HNF-6, C/EBP-α, C/EBP-β and C/EBP-γ). The synthetic heat-inducible promoter system utilizes a tetracycline-responsive transactivator (tTA) with a transcriptional positive-feedback loop and EGFP reporter, enabling stringent induction of transgenes upon transient heat treatment (43 °C, 30 min). Following induction, hi-Hep cells exhibit enhanced hepatic functions including ammonia detoxification, albumin secretion, and cytochrome P450 activity compared to parental HepG2 and HepG2/8F_HS cells. Transcriptome profiling was performed using Agilent SurePrint G3 Human GE 8×60K v3 arrays to compare hi-Hep cells with or without heat treatment, in monolayer and spheroid culture formats, under both serum-containing and optimized serum-free conditions, alongside parental HepG2/8F_HS controls.