Bevacizumab Increases Endothelin-1 Production via Forkhead Box Protein O1 in Human Glomerular Microvascular Endothelial Cells In Vitro

Figure 3

The nuclear localization of FoxO1 was increased in human glomerular microvascular endothelial cells (hGECs) treated with bevacizumab. hGECs were plated in 24-well plates (1 × 10⁵ cells/well) for ELISA or 60 mm culture dishes (1 × 10⁶ cells/dish) for RT-PCR and western blot. hGECs that were 90% confluent were serum-starved for 24 h before the experiments were performed. hGECs were treated with 1 µM bevacizumab and 0.1 µM AS1842856 (FoxO1 inhibitor) for 8 h, and western blots were used to evaluate the changes in FoxO1 protein level in cytosolic (cyto) and nuclear (nu) fractions after bevacizumab treatment (a). The levels of ET-1 mRNA (b) and ET-1 protein (c) were measured using real-time RT-PCR and the ELISA after bevacizumab and AS1842856 treatment. Data shown are means ± SD (n = 3;  < 0.05). The experiments were repeated at least three times, with reproducible results.

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