Effect of glycerol and low pH on heat-induced cell killing and loss of cellular DNA polymerase activities in Chinese hamster ovary cells.
A whole-cell assay technique for DNA polymerase alpha and beta was used to measure the activities of both enzymes in Chinese hamster ovary (CHO) cells after hyperthermic treatment of 42.2 - 45.5 degrees C in acidic or basic environment and in the presence or absence of 5% glycerol. Cell survival was measured at the same time, and the DNA polymerase activities were correlated with survival. The results show a positive correlation between cell killing by heat and loss of DNA polymerase beta activity, both when cells were sensitized to heat by treatment at pH 6.7 with or without glycerol and when cells were protected from heat by treatment with 5% glycerol at pH 7.4 or 6.7. The results show a poor correlation between loss of DNA polymerase alpha activity and cell survival; i.e., compared to cell killing, the loss of DNA polymerase alpha activity was sensitized to heat more by acidic treatment without glycerol and was protected less from heat by glycerol treatment at normal physiological pH (pH 7.4). However, cell killing and loss of polymerase alpha activity did correlate well for sensitization to heat by acidic treatment in the presence of glycerol and for protection from heat by glycerol treatment at low pH. These results considered with other hyperthermia-polymerase studies suggest that heat effects on membranes can apparently result in changes in environmental conditions within the cell (secondary effects), which can in turn alter polymerase activities and/or the direct or secondary effect of heat on the polymerase enzymes. Furthermore, loss of polymerase beta activity serves as a better index of thermal damage resulting in cell death than loss of alpha activity.
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004256:DNA Polymerase I / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism*; D005260:Female; D005990:Glycerol / Q000494:pharmacology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D010053:Ovary; D013997:Time Factors
D004256:DNA Polymerase I; D004257:DNA Polymerase II; D005990:Glycerol