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Abstract
Cell death resulting from cadmium (Cd) intoxication has been confirmed to induce both necrosis and apoptosis. The ratio between both types of cell death is dose- and cell-type-dependent. This study used the human keratinocytes HaCaT expressing a mutated p53 and the rat glial cells C6 expressing a wild p53 as models to characterize Cd-induced apoptosis, using sub-lethal and lethal doses. At these concentrations, features of apoptosis were observed 24 h after C6 cell treatment: apoptotic DNA fragmentation and caspase-9 activation, whereas Cd did not induce caspase-3. In HaCaT, Cd did not induce apoptotic DNA fragmentation or caspase-9 and -3 activation. The results also showed that the inhibition of p53 led to a resistance of the C6 cells to 20 µm Cd, decreased the apoptosis and increased the metallothioneins in these cells. p53 restoration increased the sensitivity of HaCaT cells to Cd but did not affect the MT expression. The results suggest that Cd induced apoptosis in C6 cells but a non-apoptotic cellular death in HaCaT cells.
| Abbreviations | ||
| CAPS | = | 3-cyclohexylamino-1-propanesulphonic acid |
| ROS | = | reactive oxygen species |
| Cd | = | cadmium |
| PBS | = | phosphate-buffered saline |
| ABTS | = | 2,2’-azino-di-[3-ethylbenzthiazoline-sulphonate(6)] |
| TBS-T | = | tris-buffered saline containing tween 20 |
| LEHD-pNA | = | N-acetyl-Leu-Glu-His-Asp-pNA (p-nitroanilide) |
| DMSO | = | dimethylsulphoxide |
| AU | = | apoptotic units |
| ELISA | = | enzyme-linked immunosorbent assay |
| EDTA | = | ethylenediaminetetraacetic acid |
| FBS | = | foetal bovine serum |
| BCA | = | bicinchoninic acid |
| SDS-PAGE | = | sodium dodecyl sulphate-polyacrylamide gel electrophoresis |
| BSA | = | bovin serum albumin |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
| FIENA | = | fluorometric immunosorbent enzyme assay |
| AFC | = | 7-amido-4-trifluoromethyl-coumarin |
| DEVD-AFC | = | Acetyl-Asp-Lime-Valley-Asp-7-amido-4-trifluoromethyl-coumarin |
| DTT | = | dithiothreitol |
| DMEM | = | dulbecco modified eagle's medium |
| RPMI | = | roswell park memorial institute medium |
| Abbreviations | ||
| CAPS | = | 3-cyclohexylamino-1-propanesulphonic acid |
| ROS | = | reactive oxygen species |
| Cd | = | cadmium |
| PBS | = | phosphate-buffered saline |
| ABTS | = | 2,2’-azino-di-[3-ethylbenzthiazoline-sulphonate(6)] |
| TBS-T | = | tris-buffered saline containing tween 20 |
| LEHD-pNA | = | N-acetyl-Leu-Glu-His-Asp-pNA (p-nitroanilide) |
| DMSO | = | dimethylsulphoxide |
| AU | = | apoptotic units |
| ELISA | = | enzyme-linked immunosorbent assay |
| EDTA | = | ethylenediaminetetraacetic acid |
| FBS | = | foetal bovine serum |
| BCA | = | bicinchoninic acid |
| SDS-PAGE | = | sodium dodecyl sulphate-polyacrylamide gel electrophoresis |
| BSA | = | bovin serum albumin |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide |
| FIENA | = | fluorometric immunosorbent enzyme assay |
| AFC | = | 7-amido-4-trifluoromethyl-coumarin |
| DEVD-AFC | = | Acetyl-Asp-Lime-Valley-Asp-7-amido-4-trifluoromethyl-coumarin |
| DTT | = | dithiothreitol |
| DMEM | = | dulbecco modified eagle's medium |
| RPMI | = | roswell park memorial institute medium |