Redox imbalance induced by docetaxel in the neuroblastoma SH-SY5Y cells: a study of docetaxel-induced neuronal damage

Figures

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  Figure 1.. Cell viability after incubation in the presence of different DTX concentrations (1, 5, 10, and 100 nM) for 6 h. MTT assay results show a progressive decline in live cells with increasing concentrations of DTX. The results indicated statistical differences (*P < 0.05; **P < 0.01 and, ***P < 0.001) between DTX-treated (at different doses) and control samples (ANOVA with Tukey’s post hoc test). The results were the means of three independent experiments performed in triplicate.

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  Figure 2.. Immunofluorescence staining in the neuroblastoma SH-SY5Y cells using a monoclonal anti-tubulin antibody. In panel a, untreated cells are displayed and the label shows a clear localisation in the cytoskeletal microtubules (green). In panel b, the fluorescent signal appears weak and limited in the reduced cytoplasm, losing its organisation in cells treated with DTX. The nuclei (DAPI blue-fluorescent DNA stain) appeared altered, panel b. Bar 20 μm.

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  Figure 3.. Annexin-V/Propidium Iodide assay in the neuroblastoma SH-SY5Y cells. Panel a) untreated cells; panels b–d) cells treated with DTX. Apoptotic (green, panel b), necrotic (red, panel c), and apoptotic/necrotic cells (green/red, panel d) are shown by Annexin-V/Propidium Iodide assay. Bar 20 μm.

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  Figure 4.. Transmission electron micrographs of the neuroblastoma SH-SY5Y cells. In panel a untreated cells are shown: the cells have regular nuclei (N) and organised cytoplasm. In panel b cells after DTX treatment are displayed: on the right an apoptotic cell is shown. Marginated chromatin is evident (arrow); the cytoplasm is devoid of organelles. Bar 12 μm.

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  Figure 5.. Total and free F2-IsoPs levels in DTX-treated and control cells. DTX treatment was 1.25 nM for 6 h. Data were means ± standard deviations of nine samples; **P < 0.01; ***P < 0.001 compared to control (C) samples (Student’s test). Legend: C, control (untreated) cells; DTX docetaxel, F2-IsoP, F2-isoprostanes

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  Figure 6.. Reduced and oxidised glutathione levels in DTX-treated (1.25 nM, 6 h) and control cells. Data were means ± standard deviations of nine samples. **P < 0.01; ***P < 0.001 compared to control values (Student’s test). Legend: C, control (untreated) cells; DTX docetaxel, GSH, reduced glutathione; GSSG, oxidised glutathione.

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  Figure 7.. Levels of ascorbic acid, and catalase activity in DTX-treated (1.25 nM, 6 h) and control cells. Data were means ± standard deviations of nine samples. *P < 0.05 compared to control values (Student’s test). Legend: C, control (untreated) cells; DTX docetaxel.