Heat induction of reporter gene expression via the gadd153 promoter and its possible application to hyperthermia treatment of cancer

Abstract

The use of the gadd153promoter to induce expression of a reporter geneunder heat stress conditions was investigated,since the results of previous studies have suggestedthat the gadd153promoter is likely to be activated by the indirecteffects of hyperthermia, that is, by DNA damage thatoccurs when reactive oxygen species are produced byheat stress. The optimum temperature for a significantinduction was found to be between 41 and 43 degrees C andincreased expression of the reporter gene was observedat about 24 h after the heat treatment. Under theseconditions, the cell integrity was not alteredmorphologically and the growth stopped temporarily,while the viability was maintained. A second increasein expression occurred at a later stage when the cellswere severely damaged at 43-45 degrees C. Atthese temperatures, the cellular morphology showedsignificant alteration and the growth was stronglyarrested. This is likely to be due to a differentmechanism which could involve DNA repair processes. Itis expected that this method of induction can beexploited to drive the production of a protein ofinterest in a cancer treatment program that includes hyperthermia.