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Abstract
In M07e cells, a human megakaryocytic leukaemia line, reactive oxygen species (ROS) are generated in response to cytokines acting as intracellular messengers to modulate glucose transport. The aim of this work was to study the signal cascade involved in the acute glucose transport activation in cells exposed to growth factors, such as granulocyte macrophage-colony stimulation factor (GM-CSF) and thrombopoietin (TPO), to better understand some aspects of the aberrant proliferation in leukaemia. Results confirm ROS involvement in modulation of glucose transport in this cell line. Furthermore, GM-CSF and TPO produced changes in Glut1 phosphorylation and specific inhibitors employed to identify protein kinases involved in Glut activation by these cytokines proved that Akt, PLCγ, Syk and the Src family take part in signal transduction leading to Glut1 activation.
| Abbreviations | ||
| DOG | = | 2-deoxy-D-glucose |
| BHA | = | butylated hydroxyanisole |
| BAPTA-AM | = | 1,2-bis (2-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid tetrakis (acetoxymethylester) |
| [Ca2 + ]c | = | cytosolic Ca2 + concentration |
| ER | = | endoplasmic reticulum |
| GM-CSF | = | granulocyte macrophage-colony stimulation factor |
| IL-3 | = | interleukin-3 |
| IMDM | = | Iscove's modified Dulbecco's medium |
| PBS | = | phosphate buffered saline |
| PI3K | = | phosphatidylinositol 3-kinase |
| PIC | = | piceatannol |
| PKB | = | protein kinase B |
| PLC | = | phospholipase C |
| PMSF | = | phenylmethylsulphonyl fluoride |
| ROS | = | reactive oxygen species |
| SCF | = | stem cell factor |
| SOD | = | superoxide dismutase |
| TK | = | tyrosine kinase |
| TLCK | = | N-tosyl-L-lysine chloromethyl ketone |
| TPCK | = | N-tosyl-L-phenylalanine chloromethyl ketone |
| TPO | = | thrombopoietin |
| SERCA | = | sarcoplasmic/endoplasmic reticulum calcium ATPases |
| TG | = | thapsigargin |
| Abbreviations | ||
| DOG | = | 2-deoxy-D-glucose |
| BHA | = | butylated hydroxyanisole |
| BAPTA-AM | = | 1,2-bis (2-aminophenoxy)ethane-N,N,N’,N’-tetraacetic acid tetrakis (acetoxymethylester) |
| [Ca2 + ]c | = | cytosolic Ca2 + concentration |
| ER | = | endoplasmic reticulum |
| GM-CSF | = | granulocyte macrophage-colony stimulation factor |
| IL-3 | = | interleukin-3 |
| IMDM | = | Iscove's modified Dulbecco's medium |
| PBS | = | phosphate buffered saline |
| PI3K | = | phosphatidylinositol 3-kinase |
| PIC | = | piceatannol |
| PKB | = | protein kinase B |
| PLC | = | phospholipase C |
| PMSF | = | phenylmethylsulphonyl fluoride |
| ROS | = | reactive oxygen species |
| SCF | = | stem cell factor |
| SOD | = | superoxide dismutase |
| TK | = | tyrosine kinase |
| TLCK | = | N-tosyl-L-lysine chloromethyl ketone |
| TPCK | = | N-tosyl-L-phenylalanine chloromethyl ketone |
| TPO | = | thrombopoietin |
| SERCA | = | sarcoplasmic/endoplasmic reticulum calcium ATPases |
| TG | = | thapsigargin |