Transcriptomics identifies mild hyperthermia-conditioned tumor supernatant as an inducer of HepG2 transition

Inducing the transition of tumor cells into normal hepatocyte-like cells represents a promising therapeutic strategy for hepatocellular carcinoma (HCC). However, safe and effective inducers are currently lacking, and the underlying mechanisms remain poorly understood. This study aimed to investigate whether tumor supernatant (Tsn) under mild hyperthermia conditions could induce the transition of HepG2 cells into normal hepatocyte-like cells and to elucidate the potential mechanisms involved. HepG2 cells were treated at 42.5 °C for 60 min, and the post-treatment tumor supernatant (42.5Tsn) was collected. Transcriptome sequencing was performed to identify differentially expressed genes (DEGs) between the 42.5Tsn and 37Tsn treatment groups. Compared with the 37Tsn group, 446 DEGs were identified in the 42.5Tsn group. These DEGs were enriched in pathways related to DNA damage repair, cell cycle arrest, and nuclear receptor signaling. Among the 33 core genes, tumor suppressor genes (BRCA1, PALB2, SLX4) were significantly upregulated, while nuclear transcription factors that maintain stemness, such as HEY1, were significantly downregulated. Tsn induced by mild hyperthermia demonstrates the potential to promote the transition of HepG2 cells toward normal hepatocyte-like cells while suppressing their metastatic potential. However, due to the complex composition of Tsn and the transcriptome-only level of this analysis, the specific active factors, causal regulatory mechanisms, and in vivo efficacy require further validation through component analysis, functional assays, and animal models.

The online version contains supplementary material available at 10.1007/s10616-026-01011-w.