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Figure 3 | BMC Cancer

Figure 3

From: Docosahexaenoic acid-induced apoptosis is mediated by activation of mitogen-activated protein kinases in human cancer cells

Figure 3

DHA induces mitochondrial ROS overproduction and mitochondrial dysfunction. (A-B) PA-1 cells were incubated for 1 h with or without 5 mM NAC before exposure to 40 μM DHA for 4 h. Intracellular superoxide and mitochondrial superoxide levels were detected using DHE (A) or MitoSOX (B) probes under a fluorescence microscope (right) or by flow cytometry (left), as described in Meterial and Methods (scale bar, 50 μm). (C) MitoSOX-stained H1299 and SiHa cells were exposed to 60 μM (50 μM in case of D54MG cells) DHA with 1 h of 5 mM NAC pretreatment. After 4 h of DHA exposure, the fluorescence of MitoSOX-stained cells were observed by fluorescence microscopy (scale bar, 50 μm). (D) DHA reduces MMP. PA-1 cells were stained with 25 nM TMRE, exposed to 5 mM NAC for 1 h and then DHA was added into the media followed by a further 4 h incubation. MMP was assayed by flow cytometry analysis (left). Right, data are presented as the average mean intensity fluorescence (MFI). ***, P < 0.001. Each bar represents the mean of three determinations repeated in three separate experiments. (E) Decrease in OCR by DHA treatment. PA-1 cells were seeded in 24-well XF analysis plates, and treated with 40 μM of DHA with 1 h of 5 mM NAC pretreatment. The OCR was monitored for 2 h, and calculated relatively to the vehicle control and average of five wells is shown.

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