THERMOTOLERANCE AND THE HEAT-SHOCK RESPONSE IN NORMAL HUMAN KERATINOCYTES IN CULTURE

Protective responses of normal human epidermal keratinocytes in culture, after exposure to elevated temperatures ("heat shock"), were examined. Cell viability, measured 24-48 h after a 20-min heat challenge at temperatures between 37-degrees-C and 54-degrees-C, declined sharply within a narrow 2-degrees-C-3-degrees-C range. However, conditioning with a mild thermal pretreatment (40-degrees-C or 42-degrees-C for 1 h) protected the keratinocytes against a subsequent heat challenge. This induced thermotolerance was apparent when cells were challenged at 1, 3, and 6 h after the thermal pre-treatment, but disappeared by 24 h. Heating conditions that induce thermotolerance also stimulated the synthesis of heat-shock proteins (hsp) in these cells. Inductions of prominent S-35-methionine labeled bands at 70, 78, and 90 kDa were observed. However, the increases in synthesis of these heat-shock proteins did not correlate well with thermotolerance, because large increases were also observed at certain elevated temperatures that did not produce improved survival. Keratins observed in these cells (50 and 58 kDa classes) were not induced by heat shock. The development of thermotolerance, and the induction of hsp, were both completely blocked by 3'-deoxyadenosine (cordycepin), an inhibitor of newly synthesized messenger RNA, but not by adenosine, the normal analog. While heat-inducible mRNA apparently mediate some function important for the development of thermotolerance, the nature of that role remains speculative. Overall, our findings establish the existence of a functional thermal protective mechanism in human keratinocytes that appears to require the synthesis of new mRNA.