AKR1C3 induces resistance of hepatocellular carcinoma cells to sorafenib via modulating the ROS/SIRT1/BRD4/NFIC signaling pathway

This study aimed to investigate how AKR1C3 mediates resistance to sorafenib in hepatocellular carcinoma (HCC) cells. Bioinformatics analysis was employed to screen genes specifically expressed in sorafenib resistance HCC. Single‑cell transcriptomic profiling further dissected HCC cellular heterogeneity, revealing a proliferative cancer cell population with high migration/proliferation scores, a dedifferentiated state, complex communication networks centered on hepatocytes, and increased M2 macrophage/neutrophil infiltration. Western blotting, flow cytometry, and immunohistochemistry revealed that, compared to parental Huh7 cells, sorafenib‑resistant Huh7 cells showed significantly elevated AKR1C3, Nrf2, p‑SIRT1, SHP2, p‑PI3K, CyclinA1, MMP9, and HIF1α, and reduced P22, P47, BRD4, NFIC, and PTEN. Direct ROS measurement using DCFH-DA flow cytometry further confirmed that ROS levels were significantly reduced in sorafenib resistance cells. Flow cytometry demonstrated the role of AKR1C3 in normal hepatocytes (LO 2 ). Compared to parental Huh7 cells, sorafenib resistance Huh7 cells exhibited significantly elevated expression of AKR1C3, Nrf2, p-SIRT1, SHP2, p-PI3K, CyclinA1, MMP9, and HIF1α proteins, while expression levels of P22, P47, BRD4, NFIC, and PTEN were significantly reduced. Immunofluorescence showed enhanced relative fluorescence intensity of AKR1C3 and SHP2. Furthermore, AKR1C3 promoted the proliferation, migration, invasion, and angiogenesis of Huh7 cells by mediating the ROS/SIRT1/BRD4/NFIC signaling pathway, thereby conferring sorafenib resistance in hepatocellular carcinoma cells.