Figure 9.

Differential effect of H₂O₂ with respect to CLytA-DAAO and D-Ala treatment. (A) Plasmatic membrane rupture induced by H₂O₂ in Hs766T, IMIM-PC-2, and RWP-1 cell lines from pancreatic carcinoma, SW-480 and SW-620 cell lines from colorectal carcinoma and HGUE-GB-37 and HGUE-GB-39 cell lines from glioblastoma. Cells were treated with 600 μM H₂O₂ for 24 h and cell viability was determined using Muse cell analyzer. Graph represents cell death percentage (mean ± SD) after subtracting cell death in the control untreated (n ≥ 3). (B) Variations in cell cycle distribution after H₂O₂ treatment for 48 and 72 h. Hs766T and IMIM-PC-2 pancreatic carcinoma cell lines were treated with 600 μM H₂O₂ for 48 and 72 h and cell cycle distribution was determined by flow cytometry. Graph shows the cells percentage ± SD in each phase of cell cycle after subtracting the cells percentage in the control untreated (n ≥ 3). (C) Intracellular ROS increase after H₂O₂ treatment in IMIM-PC-2 pancreatic carcinoma cell line, SW-480 colorectal carcinoma cell line and HGUE-GB-37 glioblastoma cell line. Cells were treated with 600 μM H₂O₂ for 20–120 min. Free radical production was determined through DCFH₂-DA probe and each treatment time had a control untreated that only contained the probe. Graph shows the fold change (FC) ± SD of fluorescent intensity with respect to the control (n ≥ 6). * Indicates a p-value < 0.05, ** p-value < 0.01 and *** p-value < 0.001.