Research Paper Volume 12, Issue 11 pp 11116—11138

Deoxyelephantopin induces apoptosis via oxidative stress and enhances gemcitabine sensitivity in vitro and in vivo through targeting the NF-κB signaling pathway in pancreatic cancer

Figure 2. DET induced oxidative stress, disturbed MMP and unbalanced the REDOX system in pancreatic cancer cells. (A) The DET-induced morphological changes in BxPC-3 and CFPAC-1 cells were observed using microscope. (B) Effect of DET on oxidative stress in BxPC-3 and CFPAC-1 cells were evaluated using DCFH-DA probe and observed under fluorescent microscope. (C) Effect of DET on fluorescence intensity of DCF were measured using fluorescence microplate reader. *P < 0.05, **P < 0.01 versus CTL. ##P < 0.01 versus DET (50 μM) at 2 h or DET (60 μM) at 3 h. CTL, control. (D) Effect of DET on MMP in BxPC-3 and CFPAC-1 cells were tested using JC-1 probe and evaluated using fluorescence microplate reader. **P < 0.01 versus CTL. ##P < 0.01 versus DET (30 μM) or DET (40 μM). CTL, control. (E) Effect of DET on oxidative stress were further assessed using MitoSOX and observed under fluorescent microscope. (F) Effect of DET on fluorescence intensity of MitoSOX were detected using fluorescence microplate reader. **P < 0.01 versus CTL. #P < 0.05 versus DET single treatment groups. CTL, control. (GI) Effect of DET on intracellular GSH, GSSG and the ratio of GSH to GSSG in BxPC-3 and CFPAC-1 cells were assessed using GSSG/GSH quantification kit. *P < 0.05, **P < 0.01 versus CTL. CTL, control. (J) Effect of DET on intracellular TrxR activity was measured using thioredoxin reductase assay kit. **P < 0.01 versus CTL. CTL, control. Magnification, ×100 (A), × 200 (B, E). Scale bar, 200 μm (A), 100 μm (B, E). DCFH-DA, 2’, 7’-dichlorofluorescein-diacetate. GSH, reduced glutathione. GSSG, oxidative form of glutathione. TrxR, thioredoxin reductase.