Variation of Heat Shock Protein 70 through the Cell Cycle in HL-60 Cells and Its Relationship to Apoptosis

Abstract

Cells respond to a heat shock by synthesizing heat shock proteins, which help to protect the cells from further heating. Recent results indicate that heat shock protein 70 (hsp70) may help to protect cells from apoptosis. We have used flow cytometry to investigate the relationship between constitutive and inducible hsp70 and apoptosis through the cell cycle in HL-60 cells. Specific antibodies were used to measure hsp70 in cells costained with propidium iodide. In separate samples apoptosis was measured using the TdT assay. The apoptotic cells have a subdiploid DNA content, which allows them to be identified also in the bivariate histograms of heat shock protein vs DNA content. After HL-60 cells were heated at 45.0°C for 7.5 min and incubated for various times at 37°C, many of them underwent apoptosis. The level of inducible hsp70 (hsp72) was lower in the apoptotic cells than in the nonapoptotic population, but constitutive hsp70 (hsp73) was the same in both populations. Pretreatment with sodium vanadate increased the fraction of apoptotic cells twofold, slightly increased the level of hsp72 in the nonapoptotic cells, but did not affect hsp73. These results suggest that hsp72, but not hsp73, is involved in the development or prevention of apoptosis.