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Abstract
The mechanism involved in the prosurvival effect of interleukin-3 on the human acute myeloid leukaemia cell line M07e is investigated. A decrease in intracellular reactive oxygen species (ROS) content, glucose transport activity and cell survival was observed in the presence of inhibitors of plasma membrane ROS sources, such as diphenylene iodonium and apocynin, and by small interference RNA for Nox2. Moreover, IL-3 incubation stimulated the synthesis of Nox2 cytosolic sub-unit p47phox and glucose transporter Glut1. Thus, the inhibition of ROS generation by Nox inhibitors stimulated apoptosis showing that ROS production, induced by IL-3 via Nox2, protects leukaemic cells from cell death. Also incubation with receptor tyrosine kinase inhibitors, such as anti-leukaemic drugs blocking the stem cell factor receptor (c-kit), showed similar effects, hinting that IL-3 transmodulates c-kit phosphorylation. These mechanisms may play an important role in acute myeloid leukaemia treatment, representing a novel therapeutic target.
| Abbreviations | ||
| 2-APB | = | aminoehtyl diphenyl borinate |
| Ac-DEVD-AMC | = | acetyl-Asp-Glu-Val-Asp-7-amino-4-methyl coumarin |
| Ac-IETD-AMC | = | acetyl-Ile-Glu-Thr-Asp-7-amino-4-methyl coumarin |
| Ac-LEHD-AMC | = | acetyl-Leu-Glu-His-Asp-7-amino-4-methyl coumarin |
| AML | = | acute myelogenous leukaemia |
| CaMKII | = | Ca2 +/Calmodulin-Dependent Protein Kinase II |
| DCF | = | 2′,7′-dichlorofluorescein |
| DCFH-DA | = | 2′,7′-dichlorodihydrofluorescin diacetate |
| DOG | = | 2-deoxy-D-glucose |
| DPI | = | diphenyleneiodonium chloride |
| FCS | = | foetal calf serum |
| Glut1 | = | glucose transporter 1 |
| HE | = | hydroethidine |
| IL-3 | = | interleukin-3 |
| IMDM | = | Iscove's modified Dulbecco's medium |
| InsP3 | = | inositol 1,4,5-triphosphate |
| JNK | = | Jun N-terminal kinase |
| MAPK | = | Mitogen-Activated Protein Kinase |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
| Nox | = | NAD(P)H oxidase |
| PBS | = | phosphate buffered saline |
| PI3K | = | phosphatidylinositol 3-kinase |
| PKC | = | protein kinase C |
| PLC | = | phospholipase C |
| PMSF | = | phenylmethylsulphonyl fluoride |
| ROS | = | reactive oxygen species |
| RTK | = | receptor tyrosine kinase |
| SCF | = | stem cell factor |
| TK | = | tyrosine kinase |
| TLCK | = | N-tosyl-lysine chloromethyl ketone |
| TPCK | = | N-tosyl-phenylalanine chloromethyl ketone |
| VEGFR2 | = | vascular endothelial growth factor receptor 2 |
| Abbreviations | ||
| 2-APB | = | aminoehtyl diphenyl borinate |
| Ac-DEVD-AMC | = | acetyl-Asp-Glu-Val-Asp-7-amino-4-methyl coumarin |
| Ac-IETD-AMC | = | acetyl-Ile-Glu-Thr-Asp-7-amino-4-methyl coumarin |
| Ac-LEHD-AMC | = | acetyl-Leu-Glu-His-Asp-7-amino-4-methyl coumarin |
| AML | = | acute myelogenous leukaemia |
| CaMKII | = | Ca2 +/Calmodulin-Dependent Protein Kinase II |
| DCF | = | 2′,7′-dichlorofluorescein |
| DCFH-DA | = | 2′,7′-dichlorodihydrofluorescin diacetate |
| DOG | = | 2-deoxy-D-glucose |
| DPI | = | diphenyleneiodonium chloride |
| FCS | = | foetal calf serum |
| Glut1 | = | glucose transporter 1 |
| HE | = | hydroethidine |
| IL-3 | = | interleukin-3 |
| IMDM | = | Iscove's modified Dulbecco's medium |
| InsP3 | = | inositol 1,4,5-triphosphate |
| JNK | = | Jun N-terminal kinase |
| MAPK | = | Mitogen-Activated Protein Kinase |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
| Nox | = | NAD(P)H oxidase |
| PBS | = | phosphate buffered saline |
| PI3K | = | phosphatidylinositol 3-kinase |
| PKC | = | protein kinase C |
| PLC | = | phospholipase C |
| PMSF | = | phenylmethylsulphonyl fluoride |
| ROS | = | reactive oxygen species |
| RTK | = | receptor tyrosine kinase |
| SCF | = | stem cell factor |
| TK | = | tyrosine kinase |
| TLCK | = | N-tosyl-lysine chloromethyl ketone |
| TPCK | = | N-tosyl-phenylalanine chloromethyl ketone |
| VEGFR2 | = | vascular endothelial growth factor receptor 2 |