To investigate the mechanism by which SHP2 influences ferroptosis through the regulation of signaling pathways, thereby impacting hepatocellular carcinoma. Integrate bulk RNA-seq and single-cell transcriptomic data to identify differentially expressed genes and analyze cellular heterogeneity, pseudotemporal trajectories, and cell–cell communication networks. In vitro experiments involved treating HepG2, Huh7, and Hep3B cells with the SHP2 inhibitor (PHPS1), the CREB agonist (AE-18), the EZH2 inhibitor (IN-14), and the FOXO1 inhibitor (AS1842856). Western blot analysis was performed for P-SHP2, P-PI3K, nuclear P-CREB, nuclear EZH2, nuclear FOXO1, GPX4, Nur77, NCOA4, LC-3B, and SQSTM1; immunofluorescence was used to detect the localization and intensity of nuclear FOXO1 and nuclear CREB; biochemical colorimetric assays were used to measure Fe 2 ⁺, ROS, and oxidative stress markers; flow cytometry was used to detect apoptosis; CCK-8 assay for proliferation; Scratch and Transwell assays for migration and invasion. SHP2 resists ferroptosis in hepatocellular carcinoma by regulating the CREB/EZH2/FOXO1 signaling pathway, thereby modulating the protein expression of autophagy/NCOA4/GPX4. Integrated bulk RNA-seq and single-cell transcriptomic analyses further revealed that SHP2-mediated transcriptional reprogramming and altered cellular communication within the HCC microenvironment jointly promote resistance to ferroptosis.
Yu et al. (Wed,) studied this question.