Exploring the mechanism of Naringenin in the treatment of hepatocellular carcinoma based on mRNA sequencing and experimental validation

Background Hepatocellular carcinoma (HCC) ranks among the top three causes of cancer-related mortality globally and is associated with a relatively low five-year overall survival rate. Naringenin has demonstrated significant inhibitory effects on various neoplasms; however, the mechanisms of action and potential molecular targets of naringenin in the context of HCC remain to be elucidated. Methods Cellular proliferation in cancer cells was quantified using the Cell Counting Kit-8 (CCK-8) assay. Wound healing and transwell tests were employed to evaluate the migratory and invasive capabilities of the cells, respectively. Apoptosis was evaluated using Hoechst staining to visualize nuclear changes and flow cytometry to quantify apoptotic populations. Following mRNA sequencing, we integrated the TCGA database with known naringenin-related targets to identify overlapping genes, which were subsequently subjected to clinical significance analysis. The expression of these genes was confirmed at the protein and mRNA levels using Western blot (WB) and quantitative PCR (qPCR), respectively. In vivo experiments were conducted using an MHCC-97H xenograft model in nude mice, with histopathological examination of tumor sections performed using hematoxylin and eosin (H&E) staining. Results In vitro, naringenin demonstrated a potent inhibitory effect on the proliferation, invasion, and migration of MHCC-97H and Huh7 cells while exhibiting a pronounced pro-apoptotic impact on both cell lines. mRNA sequencing results revealed significant differential gene expression. Utilizing Venn diagrams, we identified key genes, including IGFBP3, PGF, CA9, AKR1C3, KLK1, and CHRNA7. We implicated signaling pathways such as the "Wnt signaling pathway" and "MAPK signaling pathway" as potentially critical in naringenin's anti-HCC activity. The clinical significance analysis revealed that CA9 and AKR1C3 were identified as autonomous prognostic variables for hepatocellular carcinoma (HCC), a conclusion supported by molecular docking investigations. The therapeutic promise of naringenin was further supported by its considerable reduction in tumor weight and volume shown in animal trials. Conclusion This study shows that naringenin may regulate signaling pathways by targeting a series of genes: IGFBP3, PGF, CA9, AKR1C3, KLK1, and CHRNA7, resulting in the inhibition of tumor cell proliferation and metastasis, alongside the promotion of apoptosis.