Objective:To search for potential synergistic drugs that can enhance the inhibitory effect of lenvatinib on the proliferation of hepatocellular carcinoma cells by compound screening and explore the underlying molecular mechanisms.Methods:The impact of lenvatinib on the proliferation of Huh7 cells was detected using CCK-8 assay and long-term cell proliferation assays.The potential synergistic drugs for lenvatinib in Huh7 cells was screened using an FDA-approved compound library.Subsequently,the inhibitory effect of lenvatinib combined with zinc pyrithione on the proliferation of Huh7 cells was assessed using CCK-8 assay and long-term cell proliferation assay.Furthermore,RNA-sequencing was utilized to investigate the changes in gene expression profiles following the combined action of zinc pyrithione and lenvatinib,exploring the potential molecular mechanisms.The impact of combination therapy on signaling pathways was investigated through Gene Set Enrichment Analysis(GSEA)and Gene Ontology Analysis(GO).Results:Lenvatinib exerts dose-dependent inhibition on the proliferation of Huh7 in vitro(IC50 value=0.190 μmol/L).Screening of compound libraries identified zinc pyrithione as a compound that synergistically promotes the inhibition of Huh7 cell proliferation by lenvatinib.This result was further validated through CCK-8 assay(P<0.05)and long-term cell proliferation experiments.Compared to treatment with lenvatinib alone,the combination treatment of zinc pyrithione and lenvatinib upregulated signaling pathways related to oxidative stress response,apoptosis,and cellular responses to copper ions in Huh7 cells,with Gene Set Enrichment Analysis(GSEA)showing significant enrichment of the oxidative phosphorylation pathway.Conclusion:Lenvatinib can inhibit the proliferation of Huh7 cells in vitro,while Zinc Pyrithione can synergize with lenvatinib to exert a more significant inhibitory effect on the Huh7 cells.The molecular mechanism of this synergistic effect may involve generation of a large amount of reactive oxygen species(ROS)to induce apoptosis of liver cancer cells through oxidative stress response,as well as promoting the death of hepatocellular carcinoma cells by disrupting copper homeostasis.

Objective:To search for potential synergistic drugs that can enhance the inhibitory effect of lenvatinib on the proliferation of hepatocellular carcinoma cells by compound screening and explore the underlying molecular mechanisms.Methods:The impact of lenvatinib on the proliferation of Huh7 cells was detected using CCK-8 assay and long-term cell proliferation assays.The potential synergistic drugs for lenvatinib in Huh7 cells was screened using an FDA-approved compound library.Subsequently,the inhibitory effect of lenvatinib combined with zinc pyrithione on the proliferation of Huh7 cells was assessed using CCK-8 assay and long-term cell proliferation assay.Furthermore,RNA-sequencing was utilized to investigate the changes in gene expression profiles following the combined action of zinc pyrithione and lenvatinib,exploring the potential molecular mechanisms.The impact of combination therapy on signaling pathways was investigated through Gene Set Enrichment Analysis(GSEA)and Gene Ontology Analysis(GO).Results:Lenvatinib exerts dose-dependent inhibition on the proliferation of Huh7 in vitro(IC50 value=0.190 μmol/L).Screening of compound libraries identified zinc pyrithione as a compound that synergistically promotes the inhibition of Huh7 cell proliferation by lenvatinib.This result was further validated through CCK-8 assay(P<0.05)and long-term cell proliferation experiments.Compared to treatment with lenvatinib alone,the combination treatment of zinc pyrithione and lenvatinib upregulated signaling pathways related to oxidative stress response,apoptosis,and cellular responses to copper ions in Huh7 cells,with Gene Set Enrichment Analysis(GSEA)showing significant enrichment of the oxidative phosphorylation pathway.Conclusion:Lenvatinib can inhibit the proliferation of Huh7 cells in vitro,while Zinc Pyrithione can synergize with lenvatinib to exert a more significant inhibitory effect on the Huh7 cells.The molecular mechanism of this synergistic effect may involve generation of a large amount of reactive oxygen species(ROS)to induce apoptosis of liver cancer cells through oxidative stress response,as well as promoting the death of hepatocellular carcinoma cells by disrupting copper homeostasis.