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

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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. (Mol Cell Biochem 167: 145-152, 1997)

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Authors and Affiliations

  1. Department of Clinical Physiology, Division of Medicine, Walter Reed Army Institute of Research, Washington, DC, 20307-5100, USA

    X.Z. Ding, G.C. Tsokos, R.C. Smallridge & J.G. Kiang

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  1. X.Z. Ding
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  2. G.C. Tsokos
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  3. R.C. Smallridge
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  4. J.G. Kiang
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Ding, X., Tsokos, G., Smallridge, R. et al. Heat shock gene-expression in HSP-70 and HSF1 gene-transfected human epidermoid A-431 cells. Mol Cell Biochem 167, 145–152 (1997). https://doi.org/10.1023/A:1006833522418

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