The Effect of Mild Hyperthermia on Morphology, Ultrastructure and F-Actin Organization in HL-60 Cell Line
DOI:
https://doi.org/10.12775/mbs-2013-0012Keywords
hyperthermia, actin filaments, HL-60 cell lineAbstract
Introduction. Hyperthermia is a well-established physical stimulus, which is applied as an adjunctive therapy with various cancer treatments, such as radiotherapy and chemotherapy. However, the precise mechanism of heat action at the cellular level remains to be elucidated, and appears to be multi-dimensional. The purpose of the current study was to determine the effect of mild hyperthermia on the actin cytoskeleton in the HL-60 cell line. In addition, the morphological and ultrastructural approaches were used to determine the type of hyperthermia-induced cell death.
Material and methods. All studies were performed using human promyelocytic leukemia cell line (HL-60). Actin filaments were visualized with phalloidin conjugated to Alexa Fluor® 488 using fluorescence microscopy. Morphological and ultrastructural changes in the HL-60 cells were analysed by light and electron microscopy, respectively.
Results. Exposure of HL-60 cells to mild hyperthermia resulted in the reorganization of the actin cytoskeleton and the appearance of characteristic apoptotic features, including cell shrinkage, chromatin condensation and margination. In addition, swollen mitochondria were observed. The morphological and ultrastructural changes increased in severity with an increase in recovery time. Similarly, actin filament remodeling was observed immediately after the heat shock and was more evident 3 and 6 hrs after the treatment. These effects were mainly reflected by a higher definition of the dense cortical F-actin ring as well as the appearance of brightly fluorescent F-actin dots and networks scattered throughout the cytoplasm.
Conclusions. Presented data suggest that actin filament reorganization is involved in the process of apoptosis initiated by mild hyperthermia. Furthermore, the results of our studies showed that the severity of hyperthermia-induced morphological and ultrastructural changes as well as alterations in actin organization depend not only on the temperature treatment but also on the duration of post heat shock recovery
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