Study on the Bioinformatics and in Vitro Experimental Mechanism of Guben Xiaoji Decoction Regulating Copper Death in Hepatocellular Carcinoma

HOU Zongwei; LI Junxi; YANG Renyi; LIU Jingze; ZENG Puhua

doi:10.3639/j.issn.2095-3097.2025.03.028

Abstract：Objective To identify the targets and related signaling pathways of GuBen Xiaoji Decoction(GBXJD)in regulating cuproptosis in hepatocellular carcinoma(HCC)through bioinformatics analysis and validate its mechanisms via in vitro experiments.Methods Active components and targets of GBXJD were collected from The traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP)and Herb databases.Differentially expressed genes(DEGs)in HCC and cuproptosis-related genes were retrieved from the GEO and TCGA databases.Core targets were analyzed using R soft-ware.Protein-protein interaction(PPI)networks and Gene Ontology(GO)/Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed using the STRING and DAVID databases.Key genes were screened by Lasso regression.Drug-containing serum of GBXJD was prepared,and HepG2/Lo2 cells were cultured.Cells were treated with 20%blank serum or 20%drug-containing serum for 48 hours.Gene expression was validated by quantitative real-time polymerase chain reaction(qRT-PCR).Results A total of 219 active components and 794 targets of GBXJD were identified.HCC-related DEGs(827)and cuproptosis-associated genes(44 123)were intersected,yielding 64 core DEGs.PPI networks revealed 56 key targets,including MYC,EGF,EGFR,SREBF1,and LDHA.GO/KEGG analyses identified 607 biological processes and 20 pathways.Lasso regression selected six core genes(LDHA,AKR1B10,etc.),among which five(e.g.,LDHA,AKR1B10)showed favor-able prognosis in low-risk groups(P<0.05).Conclusion GBXJD may regulate cuproptosis through targets such as LDHA and AKR1B10 to induce it in HCC HepG2 cells.Further validation of the specific mechanisms is required.