Journal: Journal of Hepatocellular Carcinoma

Article Title: Identification of a Prognostic Gene Signature Based on Lenvatinib Resistance in Hepatocellular Carcinoma with Functional Validation of the Key Gene CPB2

doi: 10.2147/JHC.S579244

Figure Lengend Snippet: Construction of lenvatinib resistant cell lines and screening of genes related to drug resistance. ( a and b ) Lenvatinib-resistant cell lines Huh7-LR and PLC/PRF/5-LR were subjected to IC50 determination using the CCK8 assay; ( c – e ) Evaluation of drug resistance was conducted through cell plate cloning assay, comparing clone proliferation between parental cells and resistant cells, with results quantified in a histogram, (n = 3, error bars ± SD); ( f – h ) Apoptosis levels in resistant and parental cells were assessed via flow cytometry, with results presented in a histogram and quantified, (n = 3, error bars ± SD); ( I and j ) Volcano plots displayed differentially expressed genes in Huh7-LR and PLC/PRF/5-LR compared to their respective parent cell lines, as identified through RNA sequencing; ( k and l ) KEGG analysis revealed predominant involvement of differentially expressed genes in biological processes for Huh7-LR ( k ) and PLC/PRF/5-LR ( l ); ( m ) Volcano plots displayed differentially expressed genes in HCC tissues compared to normal tissues based on the TCGA-LIHC dataset; ( n ) A Venn diagram illustrated the overlap between differentially expressed genes in drug-resistant cell lines and HCC tissues.* p <0.05,*** p <0.001.

Article Snippet: Two types of HCC cell lines, Huh7 and PLC/PRF/5, were treated with lenvatinib (MedChemExpress, E7080) for 48 h at concentrations ranging from 0 to 80 μM to determine the half-maximal drug inhibitory concentration (IC50).

Techniques: CCK-8 Assay, Cloning, Flow Cytometry, RNA Sequencing

Journal: Journal of Hepatocellular Carcinoma

Article Title: Identification of a Prognostic Gene Signature Based on Lenvatinib Resistance in Hepatocellular Carcinoma with Functional Validation of the Key Gene CPB2

doi: 10.2147/JHC.S579244

Figure Lengend Snippet: Low CPB2 expression in HCC is associated with poor prognosis and promotes tumor progression. ( a – c ) CPB2 expression was markedly reduced in HCC compared to normal liver tissue in unpaired ( a ) and paired ( b ) HCC samples from the TCGA-LIHC dataset, as well as in clinical samples post liver cancer surgery ( c ); ( D and E ) Kaplan-Meier survival analysis investigating the association between CPB2 expression levels and overall survival ( d ), progression-free survival ( e ), and disease-specific survival ( f ) of HCC patients; ( g ) An overexpression plasmid of CPB2 was transfected into Huh7 and PLC/PRF/5 liver cancer cells, and Western blot analysis confirmed the efficacy of overexpression; ( h and i ) CCK8 assay assessing the impact of CPB2 overexpression on HCC cell proliferation (n = 3, error bars ± SD); ( j ) EdU staining evaluating the effect of CPB2 on DNA replication; ( k and l ) The migration ( k ) and invasion ( l ) abilities of HCC cells following CPB2 overexpression were assessed using Transwell assays. ** p <0.01,*** p <0.001.

Article Snippet: Two types of HCC cell lines, Huh7 and PLC/PRF/5, were treated with lenvatinib (MedChemExpress, E7080) for 48 h at concentrations ranging from 0 to 80 μM to determine the half-maximal drug inhibitory concentration (IC50).

Techniques: Expressing, Over Expression, Plasmid Preparation, Transfection, Western Blot, CCK-8 Assay, Staining, Migration

Journal: Journal of Hepatocellular Carcinoma

Article Title: Identification of a Prognostic Gene Signature Based on Lenvatinib Resistance in Hepatocellular Carcinoma with Functional Validation of the Key Gene CPB2

doi: 10.2147/JHC.S579244

Figure Lengend Snippet: CPB2 overexpression in HCC cells increases sensibility to lenvatinib. ( a ) Western blot analysis revealed low expression of CPB2 in drug-resistant cells; ( b )Huh7-LR and PLC/PRF/5-LR cell lines were transfected with an overexpression plasmid (OE-CPB2), and the efficiency of overexpression was assessed using Western blot analysis. ( c and d ) The resistance to lenvatinib in Huh7-LR and PLC/PRF/5-LR cells transfected with the CPB2 overexpression plasmid was analyzed using CCK8 assays ( e – g ) Cell cloning experiments showed that transfection of the CPB2 plasmid decreased the formation of HCC cell clones following treatment with DMSO or lenvatinib( e ), with quantitative results presented in histograms (n = 3, error bars ± SD)( f and g ); ( h – j ) Flow cytometry analysis indicated that transfection of the CPB2 plasmid increased the apoptosis ratio of HCC cells treated with DMSO and lenvatinib( h ), with quantitative results displayed in a histogram (n = 3, error bars ± SD) ( I and j ). * p <0.05, ** p <0.01,*** p <0.001.

Article Snippet: Two types of HCC cell lines, Huh7 and PLC/PRF/5, were treated with lenvatinib (MedChemExpress, E7080) for 48 h at concentrations ranging from 0 to 80 μM to determine the half-maximal drug inhibitory concentration (IC50).

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, Cloning, Clone Assay, Flow Cytometry

Journal: Journal of Hepatocellular Carcinoma

Article Title: Identification of a Prognostic Gene Signature Based on Lenvatinib Resistance in Hepatocellular Carcinoma with Functional Validation of the Key Gene CPB2

doi: 10.2147/JHC.S579244

Figure Lengend Snippet: CPB2 enhances the susceptibility of drug-resistant cells to lenvatinib through the suppression of the MAPK signaling pathway. ( a ) Gene Ontology (GO) enrichment analysis elucidated the primary biological functions associated with CPB2; ( b ) Gene Set Enrichment Analysis (GSEA) identified pathways significantly enriched in CPB2 low-expression group; ( c ) Western blot analysis confirmed the impact of CPB2 overexpression on MAPK pathway proteins in HCC cells; ( d and e ) Subcutaneous tumor formation assays in nude mice demonstrated that DDX1-overexpressing Huh7-LR cells regained sensitivity to lenvatinib; ( f and g ) Tumor growth curve were assessed in the OE-CPB2 and Vector groups following treatment with DMSO or lenvatinib; ( h and i ) Tumor weight were assessed in the OE-CPB2 and Vector groups following treatment with DMSO or lenvatinib. * p <0.05, ** p <0.01,*** p <0.001.

Article Snippet: Two types of HCC cell lines, Huh7 and PLC/PRF/5, were treated with lenvatinib (MedChemExpress, E7080) for 48 h at concentrations ranging from 0 to 80 μM to determine the half-maximal drug inhibitory concentration (IC50).

Techniques: Expressing, Western Blot, Over Expression, Plasmid Preparation