Exploration of the inhibitory mechanism of triptolide on hepatocellular carcinoma

WANG Yiming; ZHAO Lili; LIU Wenjing; SUN Xiangjun

doi:10.13431/j.cnki.immunol.j.20260013

Abstract：Objective To investigate the antitumor mechanism of triptolide(TPL)in liver hepatocellular carcinoma(LIHC).Methods Potential targets of TPL were screened through network pharmacology and transcriptomic analysis based on TCMSP,SuperPred,TCGA-LIHC and AnimalTFDB databases,and molecular docking was used to verify its binding ability to EZH2.RNA sequencing data from TCGA-LIHC and TPL-treated Huh7 cells were integrated for differential expression,correlation,prognostic and GSEA analyses.ChIP-Atlas and ATAC-seq data were used to evaluate the epigenetic regulatory characteristics of CSRNP1.Results EZH2 was identified as a direct binding target of TPL(binding energy=-7.186 8 kcal/mol),and there was a significant negative correlation between EZH2 and CSRNP1 in LIHC patients(r=-0.37,P＜0.001).After treatment of Huh7 cells with 50 nmol/L TPL for 48 h,the expressions of EZH2 were significantly downregulated(P≤0.01),while the expression of CSRNP1 was significantly upregulated(P＜0.01).GSEA showed that EZH2 activated E2F target gene pathway,G2M checkpoint pathway and MYC target gene pathway(all P＜0.01),and high expression of CSRNP1 inhibited MYC signaling(NES=-1.47,P=0.009).Epigenetic analysis confirmed that EZH2 silenced CSRNP1 transcription through H3K27me3 modification:in LIHC cells,the promoter region of CSRNP1 was enriched with H3K27me3 modification signal peaks and lacked ATAC-seq signals,suggesting transcriptional repression.In normal liver tissues,there was no H3K27me3 modification in this region,and the ATAC-seq signal was significant with high chromatin accessibility,which was conducive to normal gene transcription.Conclusion TPL relieves the epigenetic repression of CSRNP1 by inhibiting EZH2,suppresses the MYC pathway and alleviates the immune microenvironment.These findings reveal that the EZH2/CSRNP1/MYC axis is an important mechanism underlying the anti-hepatocarcinoma effect of TPL,providing a potential target for hepatocarcinoma treatment.