A possibility for new evaluating method of cytotoxicity by using heat shock protein assay

To determine whether heat shock proteins can be utilized as a biomarker for cytotoxicity of dental materials the induction of synthesis of heat shock proteins by mercuric chloride was examined. To analyse the synthesis of heat shock proteins, HeLa cells were labelled with [35S] methionine, and the labelled proteins were separated by SDS-PAGE and autoradiographed. Incubation of the cells in a medium containing 1.25 to 40 microM mercuric chloride markedly increased the synthesis of HSP70. At 20 or 40 microM mercuric chloride in medium, HeLa cells synthesized HSP70 at 2 h after exposure, maximally at 4-7 h, and gradually diminished thereafter. Examination of the cytotoxicity of mercuric chloride by the conventional neutral red uptake assay revealed a reduction of uptake of the dye in the presence of mercuric chloride at 40 microM and above. These findings suggest that the induction of synthesis of HSP70 is one of the most sensitive cellular responses caused by mercury ion, and the heat shock protein assay can be utilized for evaluation of the cytotoxicity of dental materials.