Fig. 1.

2-DG inhibits but lonidamine potentiates the oncolytic effect of M1 virus. a, b M1 treatment upregulated glycolysis and hypoxia pathways. The HCT 116 cell line was treated with M1 virus (MOI = 1 pfu/cell) for 24 h, and was then subjected to RNA-seq. GSEA of hallmark glycolysis and hypoxia gene sets is shown. Normalized enrichment score (NES), p, and false discovery rate (FDR) values are indicated. c Heatmap of gene expression in the glycolysis gene set. d The HCT 116 and HCT-8 cell lines were treated with vehicle, 2-DG (2.5 mM), lonidamine (50 μM), M1 virus (MOI = 1 pfu/cell), or M1 virus plus 2-DG/lonidamine for 48 h, and cell viability was detected by MTT. n = 3. Statistical analysis was performed by one-way ANOVA with Dunnett’s tests for pairwise comparisons. e The HCT 116 cell line was treated with M1 virus (MOI = 1 pfu/cell) or M1 virus plus 2-DG (2.5 mM)/lonidamine (50 μM) for 24 or 48 h. Phase contrast and fluorescence micrographs are shown. The results shown are one representative result from three similar replicate experiments. Scale bar, 100 μm. f The HCT 116 and HCT-8 cell lines were treated with vehicle, 2-DG, lonidamine, M1 virus (MOI = 1 pfu/cell), or M1 virus plus 2-DG/lonidamine for 24 h, and the infection rate of M1 virus (GFP percentage) was determined by flow cytometry. n = 3. Statistical analysis was performed by one-way ANOVA with Dunnett’s tests for pairwise comparisons. The error bars indicate the mean ± SD values from three independent experiments. ns nonsignificant; *p < 0.05, **p < 0.01, ***p < 0.001