Heat shock gene-expression in HSP-70 and HSF1 gene-transfected human epidermoid A-431 cells.

Abstract

Thermotolerant cells display attenuated heat shock protein 70 kD (HSP-70) gene expression and signal transduction such as intracellular Ca2+ concentration and inositol trisphosphate in response to sublethal heat. To further investigate the regulation of heat shock gene expression, we developed constructs containing human HSP-70 and HSF1 genes and transfected human epidermoid A-431 cells. These cells were chosen because skin cells are especially vulnerable to heat shock and other environmental stressors. We report that A431 cells can be successfully transfected with HSP-70 and HSF1 genes as shown by the elevated levels of respective message and protein. Overexpression of HSP-70 in cells transfected with HSP-70 gene led to a down-regulation of the HSF1 gene expression. Interestingly, transfection of cells with the HSF1 gene was not associated with increased expression of HSP-70. Exposure of HSF1 gene-transfected cells to heat resulted in a transient but significant increase in HSP-70 gene expression as compared to that found in vector-transfected cells, which was completely inhibited by treatment with staurosporine. In conclusion, we have demonstrated successful transfection of human A-431 cells with HSF1 and HSP-70 genes, where the regulation of their expression can be studied.