Quantum dot imaging for HSP70 and HSF-1 kinetics in SCC-25 cells with or without leucine deprivation following heat shock

Abstract

The aim of this study was to develop a quantum dot-based approach for heat shock protein 70 (HSP70) and heat shock factor 1 (HSF-1) kinetics following heat shock, and to discover approaches to thermotherapy based on disrupting the effect of activation of HSF-1 and the accumulation of HSP70 by leucine deprivation. SCC-25 cells cultured with limiting leucine or normal leucine were stressed at 42°C for 30 min, and were cultured for 2, 4, 6, 8 and 10 h, respectively. The expression of HSP70 and HSF-1 was observed using confocal laser microscopy and semi-quantitative analysis was performed by Image-Pro Plus. At 6 h after heating, HSF-1 in cells cultured with normal leucine was activated and translocated from the cytosol to the nucleus, and the synthesis of HSP70 reached the maximum value and had a tendency to gather in the nucleus. However, in cells cultured with limiting leucine, HSF-1 activity decreased and accumulation of HSP70 was not found. Leucine deprivation results in the inactivation of HSF-1 leading to slight accumulation of HSP70 and no tendency to gather in the nucleus. Thus, HSF-1 may serve as a novel therapeutic target in the treatment of oral cancer.