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. 2020 Mar 11;12(3):655. doi: 10.3390/cancers12030655

Figure 4.

Figure 4

Glycolytic metabolism regulated cancer stemness of HBx-expressing MHCC-97H cells and LCSCs. MHCC-97H cells without or with HBx-expressing were transiently transfected with pcDNA3.1 or pcDNA3.1-HBx (1 μg/mL) for 8 h, followed by treatment with 2-DG (4 μmol/L) or STF-31 (8 μmol/L) or not for another 24 h. (A,B) The expression levels of cancer stemness-related proteins (A) and the mRNA levels of cancer stemness-related genes (B) in HBx-expressing MHCC-97H cells treated with 2-DG or not. (C,D) The expression levels of cancer stemness-related proteins (C) and the mRNA levels of cancer stemness-related genes (D) in HBx-expressing MHCC-97H cells treated with STF-31 or not. The target gene transcription was normalized to ACTB. (E,F) LCSCs were enriched by sphere-formation assay in MHCC-97H cells. The expression levels of cancer stemness-related proteins in HBx-expressing sphere-formed LCSCs treated with STF-31 (E) or 2-DG (F) or not. The gray value of band was assessed by image-pro plus 6.0. The relative expression level was shown. (G) Percentage of the sorted SP cells (R1 gate) in HBx-expressing MHCC-97H treated with 2-DG (Upper) or STF-31 (Lower) or not were detected by FCM (Left). Quantitative results were shown in bar graph (Right). SP: side population. R1 gate represented SP cells. * P < 0.05 as compared with pc3.1+Vehicle group, # P < 0.05 as compared with pc3.1-HBx+Vehicle group. pc3.1: pcDNA3.1 transfection without HBx-expressing. pc3.1-HBx: pcDNA3.1-HBx transfection with HBx-expressing.