Effect of M4N on glutathione metabolism and ROS (reactive oxygen species) production.
A: Effect of M4N on the contents of glutathione-related metabolites. This data is a selection from S1 Table. LNCaP, AsPC-1, and L428 cells were treated with M4N for 8 h and metabolite contents of the samples were measured by LC/GC mass spectroscopy by Metabolon. 'M/C' indicates the ratio of metabolite contents for the samples treated with M4N vs. the control. For paired comparisons between the control and M4N-treated samples, strongly shaded cells indicate statistically significant differences at p≤0.05 while lightly shaded cells indicate them at 0.05B: Effect of M4N on the expression of CHAC1. The LNCaP cells were treated with M4N (80 μM) for either 5 or 18 h and the expression of the protein was measured by the western blotting. β-Actin was used as the control. C: A schema for M4N-regulation of glutathione metabolism. The upward pointing arrows indicate that the contents of the metabolites associated with these arrows were significantly (p≤0.05) induced by M4N in at least two out of three cell lines (LNCaP, AsPC-1, and L428 cells) under the additional condition that the effect of M4N on the contents of these metabolites in the third cell line was an induction without statistical significant difference, no significant change, or not examined. Meanwhile the downward pointing arrows indicate that the contents of the metabolites associated with these arrows were significantly (p≤0.05) suppressed by M4N in at least two out of three cell lines (LNCaP, AsPC-1, and L428 cells) under the additional condition that the effect of M4N on the contents of these metabolites in the third cell line was a suppression without statistical significant difference, no significant change, or not examined. The exception to these rules was cystine (whose content was statistically significantly induced by M4N only in LNCaP cells. In other two cell lines the data are not available). The effect of M4N on ATP content in the whole cells is shown in Fig 6A. 'N.A.' indicates that the data on these particular metabolites are not available. The blue arrow indicates that CHAC1 expression was enhanced by M4N treatment (Fig 6B). The effect of M4N on ATP content in the whole cells is shown in Fig 6A. D: Effect of combination treatments on ROS production in LNCaP cancer cells. The cells were treated with M4N (80 μM), etoposide (30 μM), rapamycin (30 μM), or UCN-01 (2 μM). ROS content was measured at 5 h after treatment. C: control, E: etoposide, R: rapamycin, U: UCN-01, M: M4N, ME: M4N+etoposide, MR: M4N+rapamycin, MU: M4N+UCN-01.
