Resveratrol-Induced Downregulation of NAF-1 Enhances the Sensitivity of Pancreatic Cancer Cells to Gemcitabine via the ROS/Nrf2 Signaling Pathways

Figure 1

Resveratrol suppresses the level of NAF-1 and enhances the expression of Nrf2 by inducing the accumulation of ROS, which contributed to cell death in the Panc-1 and Mia paca-2 cells. (a–d) The pancreatic cancer cells were treated with increasing doses of RSV (0, 50, 100, 150, and 200 μM) for 24 h, and the accumulation of ROS was detected using 2, 7-dichlorofluorescin diacetate (DCFH-DA) probes. Representative flow cytometric pictures showing the mean fluorescence intensity (MFI) in each group were generated to create a composite image. (e–h) Indicated groups of pancreatic cancer cells were treated with RSV, H₂O₂, and NAC. Western blotting was performed to detect NAF-1, Nrf2, and the apoptosis-related proteins (Bax and Bcl-2). (i, j) Immunofluorescence staining of Nrf2 showed translocation of Nrf2 under the resveratrol and ROS stimulation in Panc-1 and Mia paca-2 cells (magnification, 400x; scale bar, 20 μm). (k–n) After the transfection with siNrf2 or siControl for 48 h, the Panc-1 and Mia paca-2 cells were treated with resveratrol for 24 h, and the protein levels of NAF-1, Nrf2, and the apoptosis-related proteins (Bax and Bcl-2) were detected by Western blotting. All the images represent the results of three independent experiments. Quantified histograms of the Western blots were shown alongside. Column: mean; bar: SD. compared with NAF-1 control group.

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