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Kiang JG, Ding XZ, McClain DE: Overexpression of HSP-70 attenuates increases in [Ca2+] i and protects human epidermoid A-431 cells after chemical hypoxia. Toxicol Appl Pharmacol. 1998 Apr;149(2):185-94.
This laboratory previously reported that thermotolerance diminishes the NaCN-induced increase in intracellular free calcium concentrations ([Ca2+] i) in human epidermoid A-431 cells and that blocking this increase protects the cells from NaCN toxicity. In this study, we report that cell viability after exposure to NaCN (10 mM, 1 h) is enhanced by the overexpression of HSP-70 resulting from heat shock (45 degrees C, 10 min), treatment with a protein kinase C activator phorbol 12 myristate 13-acetate (PMA; 1 microM, 4 h), or HSP-70 cDNA transfection. Because the toxicity of NaCN is mediated by increases in [Ca2+] i, we sought to determine whether the overexpression of HSP-70 might protect the cells by altering the [Ca2+] i response induced by NaCN. Basal [Ca2+] i in vector-, HSF1 cDNA-, and HSP-70 cDNA-transfected cells was 114 +/- 11 (n = 11), 95 +/- 5 (n = 6), and 151 +/- 11 (n = 15) nM, respectively, suggesting that HSP-70 metabolism is associated with maintenance of resting [Ca2+] i. Removal of external Ca2+ reduced the resting [Ca2+] i in all of these cells. With external Ca2+ reduced the resting [Ca2+] i by 97 +/- 21% in vector-transfected cells and 111 +/- 5% in HSF1 vector-transfected cells but by only 27 +/- 8% in HSP-70 cDNA-transfected cells. Heat shock or PMA treatment of vector- or HSF1 cDNA-transfected cells to induce HSP-70 also attenuated the NaCN-induced increase in [Ca2+] i, perhaps because of a decrease in Vmax for the uptake of external Ca2+. Removal of external Ca2+ or treatment with inhibitors of Na+/Ca2+ exchangers eliminated the NaCN-induced increase in [Ca2+] i in HSP-70 cDNA-transfected cells, but ryanodine treatment did not. HSP-70 cDNA transfection also reduced Ca2+ mobilization stimulated by various Ca (2+)-mobilizing agents. The results suggest that HSP-70 overexpression protects cells from NaCN cytotoxicity, perhaps by attenuating the [Ca2+] i response. |
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