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. 2012 Mar 1;106(7):1288–1296. doi: 10.1038/bjc.2012.66

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Copyright © 2012 Cancer Research UK

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Melatonin specifically induces cell death in a panel of Ewing's sarcoma cell lines. (A) TC-71, A673 (EWS-FLI1 type 1), SK-ES1 (EWS-FLI1 type 2) and A4573 (EWS-FLI1 type 3) cell number, determined by MTT assay decreased after 72 h of treatment with melatonin in a dose-dependent manner; ^*P<0.05 vs vehicle-treated group. There are also statistical differences in the effect of melatonin between the EWS-FLI1 type 1 cells and EWS-FLI1 type 2, 3 cells (^#P<0.05). (B) Decrease in the number of cells in Ewing's sarcoma (TC-71, SK-ES1 and A4573). A rise in cellular debris is observed under phase contrast microscopy after 72 h of melatonin treatment. Bars: 50 μm. (C), Increase of cell death in Ewing's cells was evaluated by the release of LDH into the extracellular medium, after 72 h of incubation with melatonin; ^*P<0.05 vs vehicle-treated cells; ^#P<0.05 vs EWS-FLI1 type 1 cells. (D) Increase in caspase-3 activity in a panel of Ewing's cell lines determined after 48 h of incubation with melatonin; ^*P<0.05 vs control groups. (E) Fas and Fas L mRNA expression was evaluated in the same cells by quantitative PCR. GAPDH was used as a housekeeping gene. Relative gene expression is represented as the n-fold increase compared with basal level (dotted lines represent vehicle-treated cells). ^*P⩽0.05 vs vehicle-treated cells.