title: Role of intracellular free Ca(II) and Zn(II) in dexamethasone-induced apoptosis and dexamethasone resistance in human leukemic CEM cell lines.
abstract: The levels of intracellular free Ca(II) and Zn(II) during dexamethasone (dex)-induced apoptosis in CEM cell lines were determined by 19F nuclear magnetic resonance (NMR), using the fluorinated intracellular chelator 1,2-bis-(2- amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid (5-FBAPTA). The effects of these divalent metal ions on growth rate and DNA degradation were evaluated. Measurements were done on one dex-sensitive (CEM-C7) and three different dex-resistant variants (CEM-C1, CEM-4R4, and CEM-ICR27). Dex caused a continuous increase in the Ca(II) level in dex-sensitive CEM-C7 cells, while in CEM-C1 cells dex caused an initial increase in the Ca(II) level which in approximately 36 h was restored to its normal value. The intracellular Ca(II) level in CEM-4R4 cells was not significantly affected by dex, while that of CEM-ICR27 cells decreased after dex incubation. Only the dex-sensitive CEM-C7 cells showed dex-induced DNA degradation. An intracellular free Zn(II) level of approximately 1 nM was measured for the dex-resistant CEM-C1 cells. No detectable level of intracellular Zn(II) was found in the other cell lines. Incubation with < 100 microM Zn(II) did not inhibit dex-induced apoptosis in CEM-C7 cells (e.g., DNA degradation). Treatment with approximately 250 microM Zn(II) caused significant decrease in growth rate in all cell lines and prevented dex-induced DNA degradation in CEM-C7 cells. A calibrated amount of Ca(II) ionophore (A23187), used to increase Ca(II) concentrations up to the dex-induced levels, did not induce DNA degradation in CEM-C7 or CEM-C1 cells. While elevation of intracellular Ca(II) by itself is not sufficient to initiate apoptosis in CEM-C7 cells, the results reported here suggest that Ca(II) is involved in the killing mechanism as a secondary factor. The combination of dex and ionophore caused significant DNA degradation in CEM-C1 cells, which normally showed resistance to each compound individually. The combination of dex and the Zn(II) chelator phenanthroline also caused extensive DNA degradation in the normally dex-resistant CEM-C1 cells, suggesting that Zn(II) plays a role in the dex resistance of these cells.
authors.affiliation: Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77555-0653.
authors.forename: F
authors.initials: F
authors.lastname: Adebodun
authors.forename: J F
authors.initials: JF
authors.lastname: Post
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D002118:Calcium / Q000378:metabolism*; D004273:DNA, Neoplasm / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology*; D004351:Drug Resistance; D004533:Egtazic Acid / Q000031:analogs & derivatives; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D007938:Leukemia / Q000473:pathology* / Q000503:physiopathology; D009682:Magnetic Resonance Spectroscopy; D014407:Tumor Cells, Cultured; D015032:Zinc / Q000378:metabolism*
chemical_list: D004273:DNA, Neoplasm; D004533:Egtazic Acid; D003907:Dexamethasone; C039877:5,5'-difluoro-1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D015032:Zinc; D002118:Calcium