Figure 4
From: Anti-angiogenic potential of an ethanol extract of Annona atemoya seeds in vitro and in vivo
EEAA decreases VEGF expression in tumor cells under hypoxic conditions. (A) Effect of EEAA on tumor cell growth. HCI-H460 cells were treated with EEAA (10 μ g/mL), and cell proliferation (left panel) and viability were determined after 48 h. (B) Reduced production of VEGF due to EEAA. NCI-H460 cells were cultured under normoxic (20% O2) or hypoxic (1% O2) conditions in the absence (M) and the presence of vehicle (DMSO, V) or 10 μ g/mL EEAA (EA). After 8 h of incubation, an immunoassay was performed to quantify VEGF secreted from NCI-H460 cells. (C) Down-regulation of VEGF at the transcriptional level by EEAA. NCI-H460 cells were cultured under normoxic (20% O2) or hypoxic (1% O2) conditions in the presence of increasing concentrations of EEAA (1–100 μ g/mL). After 24 h, intracellular VEGF mRNA levels were quantified using RT-PCR. The relative mRNA contents were determined based on comparison with vehicle (DMSO). The data are presented as the mean or relative mean ± S.D. of at least three independent experiments. *P < 0.05 and ***P < 0.001 compared with the vehicle groups.