Hyperthermia inhibits hypoxia-induced epithelial-mesenchymal transition in HepG2 hepatocellular carcinoma cells

Figure 1 Morphologic changes after hyperthermia or hypoxia treatment in HepG2 cells. A: Cells were cutured in 21% O₂ for 72 h; B: Cells were cultured in 1.0% O₂ for 72 h; C: Cells were treated with hyperthermia at 43 °C for 0.5 h, followed by treatment with normoxia; D: Cells were treated with hyperthermia at 43 °C for 0.5 h, followed by treatment with hypoxia.

Figure 2 Immunofluorescence assay of E-cadherin in HepG2 cells. A: Untreated cells; B: Hypoxia-treated cells; C: Hyperthermia-treated cells; D: Hyperthermia + hypoxia-treated cells.

Figure 3 Western blot analysis of E-cadherin and vimentin expression in HepG2 cells. Lanes 1, 2, 3 and 4 represent untreated cells, hypoxia-treated cells, hyperthermia-treated cells and hyperthermia + hypoxia-treated cells, respectively. The bar graphs show the relative expression levels of indicated proteins after normalization to β-actin. ^aP < 0.05 vs untreated cells; ^cP < 0.05 vs hypoxia-treated cells.

Figure 4 Western blot and reverse transcription-polymerase chain reaction analysis of Snail expression in HepG2 cells. A: Western blot: Lanes 1, 2, 3 and 4 represent untreated cells, hypoxia-treated cells, hyperthermia-treated cells and hyperthermia + hypoxia-treated cells, respectively. The bar graphs show the relative expression levels of Snail after normalization to β-actin; B: Reverse transcription-polymerase chain reaction: The representation of the lanes was the same as in (A); the bar graphs show the relative expression levels of Snail normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). ^aP < 0.05 vs untreated cells; ^cP < 0.05 vs hypoxia-treated cells.

Figure 5 Migration assay of HepG2 cells treated with hyperthermia or hypoxia. Quantification of the cell migration results from three experiments: each bar (hypoxia-, hyperthermia-, hyperthermia + hypoxia-treated cells) represents the relative levels observed in untreated cells (control). ^aP < 0.05 vs untreated cells; ^cP < 0.05 vs hypoxia-treated cells.