Apoptosis and Metabolic Reprogramming by <scp>RHART</scp> in Hepatocellular Carcinoma Through the <scp>CPSF6</scp> / <scp>MCT4</scp> /c‐Myc Signaling In Vitro and In Vivo

Though Rhus verniciflua and Artemisia capillaris have been traditionally prescribed for liver ailments, the molecular antitumor mechanism of their ethanol extract (RHART) remains undefined in hepatocellular carcinoma (HCC). Thus, this study aimed to elucidate the apoptotic mechanisms of RHART, focusing on its regulation of oncogenic drivers-cleavage and polyadenylation specificity factor 6 (CPSF6), monocarboxylate transporter 4 (MCT4), and c-Myc-and glycolysis-related pathways. RHART quality was standardized by HPLC fingerprinting. Cytotoxicity, apoptosis, and protein expression were analyzed using MTT, cell cycle, and Western blot assays in RHART-treated Hep3B and Huh7 cells. Functional rescue assays were conducted by overexpressing CPSF6 or c-Myc, while CPSF6-MCT4 interaction was confirmed by immunoprecipitation. The in vivo efficacy of RHART was validated using Hep3B xenografts in BALB/c nude mice followed by immunohistochemistry. RHART treatment induced pronounced cytotoxicity and apoptosis, as indicated by an elevated sub-G1 population and cleavage of PARP and caspase-3. RHART markedly downregulated CPSF6, MCT4, c-Myc, and glycolytic enzymes (HK2, PKM2, LDHA), thereby reducing glucose utilization and lactate output. CPSF6 directly interacted with MCT4, and overexpression of CPSF6 or c-Myc counteracted the capacity of RHART to induce apoptosis and metabolic reprogramming. In vivo, RHART significantly suppressed tumor growth and expression of CPSF6, MCT4, PCNA, and PKM2, while enhancing cleaved caspase-3. These findings provide scientific evidence that RHART induces apoptotic and metabolic reprogramming effects in hepatocellular carcinoma by targeting the CPSF6/MCT4/c-Myc signaling axis, underscoring its potential as a novel natural therapeutic candidate for liver cancer.