ALA-mediated photodynamic effect on apoptosis induction and secretion of macrophage migration inhibitory factor (MIF) and of monocyte chemotactic protein.

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ALA-mediated photodynamic effect on apoptosis induction and secretion of macrophage migration inhibitory factor (MIF) and of monocyte chemotactic protein (MCP-1) by colon cancer cells in normoxia and in hypoxia-like conditions in vitro Aleksandra Kawczyk-Krupka, MD, PhD, Andrzej M. Bugaj, Wojciech Latos, Katarzyna Wawrzyniec, Piotr Oleś, Anna Mertas, Zenon Czuba, Wojciech Król, Karolina Sieroń-Stołtny, Aleksander Sieroń Photodiagnosis and Photodynamic Therapy Volume 12, Issue 1, Pages 27-35 (March 2015) DOI: 10.1016/j.pdpdt.2014.12.013 Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 1 Cell survival of SW480 line for different concentrations of ALA: 0μM, 500μM, 1000μM and 1500μM and different light fluences: 0J/cm2, 10J/cm2, 30J/cm2 and 60J/cm2 in the absence of cobalt(II) chloride (CoCl2) (normoxia conditions) in environment determined by MTT assay as percentage of survival of control group. The values represent the mean±SD of four independent experiments (n=16). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 2 Cell survival of SW480 line for different concentrations of ALA: 0μM, 500μM, 1000μM and 1500μM and different light fluences: 0J/cm2, 10J/cm2, 30J/cm2 and 60J/cm2 in the presence of cobalt(II) chloride (CoCl2) (hypoxia-like conditions) in environment determined by MTT assay as percentage of survival of control group. The values represent the mean±SD of four independent experiments (n=16). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 3 Cell survival of SW620 line for different concentrations of ALA: 0μM, 500μM, 1000μM and 1500μM and different light fluences: 0J/cm2, 10J/cm2, 30J/cm2 and 60J/cm2 in the absence of cobalt(II) chloride (CoCl2) (normoxia conditions) in environment determined by MTT assay as percentage of survival of control group. The values represent the mean±SD of four independent experiments (n=16). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 4 Cell survival of SW620 line for different concentrations of ALA: 0μM, 500μM, 1000μM and 1500μM and different light fluences: 0J/cm2, 10J/cm2, 30J/cm2 and 60J/cm2 in the presence of cobalt(II) chloride (CoCl2) (hypoxia-like conditions) in environment determined by MTT assay as percentage of survival of control group. The values represent the mean±SD of four independent experiments (n=16). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 5 Apoptosis of SW480 line determined in fluorescence microscopy as percentage of apoptotic cells in group. (A) Normoxia conditions, control, (B) normoxia conditions, light 10J/cm2, (C) normoxia conditions, ALA 100μM, (D) normoxia conditions, ALA 100μM, light 10J/cm2, (E) hypoxia-like conditions, control, (F) hypoxia-like conditions, light 10J/cm2, (G) hypoxia-like conditions, ALA 100μM, (H) hypoxia-like conditions, ALA 100μM, light 10J/cm2. The values represent the mean±SD of three independent experiments performed in duplicate (n=6). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 6 Apoptosis of SW620 line determined in fluorescence microscopy as percentage of apoptotic cells in group. (A) Normoxia conditions, control, (B) normoxia conditions, light 10J/cm2, (C) normoxia conditions, ALA 100μM, (D) normoxia conditions, ALA 100μM, light 10J/cm2, (E) hypoxia-like conditions, control, (F) hypoxia-like conditions, light 10J/cm2, (G) hypoxia-like conditions, ALA 100μM, (H) hypoxia-like conditions, ALA 100μM, light 10J/cm2. The values represent the mean±SD of three independent experiments performed in duplicate (n=6). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 7 The concentration of MCP-1 in the supernatants from colon cancer cell culture of SW480 and SW620 line, (A) Normoxia conditions, control, (B) normoxia conditions, light 10J/cm2, (C) normoxia conditions, ALA 100μM, (D) normoxia conditions, ALA100μM, light 10J/cm2, (E) hypoxia-like conditions, control, (F) hypoxia-like conditions, light 10J/cm2, (G) hypoxia-like conditions, ALA 100μM, (H) hypoxia-like conditions, ALA 100μM, light 10J/cm2. The values represent the mean±SD of three independent experiments (n=12). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions

Figure 8 The concentration of MIF in the supernatants from colon cancer cell culture of SW480 and SW620 line. (A) Normoxia conditions, control, (B) normoxia conditions, light 10J/cm2, (C) normoxia conditions, ALA 100μM, (D) normoxia conditions, ALA 100μM, light 10J/cm2, (E) hypoxia-like conditions, control, (F) hypoxia-like conditions, light 10J/cm2, (G) hypoxia-like conditions, ALA 100μM, (H) hypoxia-like conditions, ALA 100μM, light 10J/cm2. The values represent the mean±SD of three independent experiments (n=12). Photodiagnosis and Photodynamic Therapy 2015 12, 27-35DOI: (10.1016/j.pdpdt.2014.12.013) Copyright © 2015 Elsevier B.V. Terms and Conditions