Integration of transcriptomics and proteomics uncovers novel targets underlying the protective effects of Nrf2 knockout in HEI-OC1 cells

Abstract

Abstract Cisplatin, a utilized anticancer drug in clinical practice, induces sensorineural hearing loss (SNHL) in patients. However, the precise mechanism underlying cisplatin-associated ototoxicity remains unknown. HEI-OC1 cells are immortalized cells derived from the organs of Corti mice and nuclear factor erythroid 2-related factor 2 （Nrf2） knockout (KO) significantly enhances cisplatin resistance in these cells. The exploration of transcriptomic and proteomic data from Nrf2 KO has significant implications for the identification of novel targets to enhance HEI-OC1 cisplatin resistance in Nrf2 KO and for understanding the biological characteristics associated with SNHL. The RNA-seq analysis revealed a significant enrichment of differentially expressed genes (DEGs) in the Nrf2 KO model within key signaling pathways, including the PI3K-Akt, MAPK, as well as Glutathione metabolism signaling pathways. Notably, expression levels of 17 specific genes were confirmed by RT-qPCR (Real-time Quantitative-PCR). The marker-based quantitative proteomics analysis revealed significant enrichment of differentially expressed proteins (DEPs) in key signaling pathways, including the p53, Glutathione metabolism, Ferroptosis, Platinum drug resistance signaling pathways. Furthermore, six proteins were validated by Western blotting (WB). Correlation analysis between transcriptomics and proteomics data demonstrated, focusing primarily on p53, Glutathione metabolism, and Ferroptosis signaling pathways. Moreover, 1 gene was randomly selected for RT-qPCR and WB. The biomarkers identified in this study may be key to understanding the biological mechanism by which Nrf2 KO strongly increases HEI-OC1 cisplatin resistance, and by targeting the PI3K-Akt, MAPK, Glutathione metabolism signaling pathways provide new ideas for the prevention and treatment of cisplatin-induced SNHL.