Hyperthermia induces cytoskeletal alterations and mitotic catastrophe in p53-deficient H1299 lung cancer cells

• Published in: Acta histochemica 2013-01, Vol.115 (1), p.8-15
• Main Authors: Pawlik, Andrzej, Nowak, Jakub Marcin, Grzanka, Dariusz, Gackowska, Lidia, Michalkiewicz, Jacek, Grzanka, Alina
• Format: Article
• Language: English
• Subjects: Actin; Actins - analysis; Actins - metabolism; Cell Death; Cell Survival; Cytoskeleton; Cytoskeleton - metabolism; Cytoskeleton - pathology; Fluorescent Antibody Technique; H1299 cells; Hot Temperature; Humans; Hyperthermia; Hyperthermia, Induced; Mitosis; Mitotic catastrophe; Stress Fibers - chemistry; Stress Fibers - metabolism; Stress Fibers - pathology; Tubulin - analysis; Tumor Cells, Cultured; Tumor Suppressor Protein p53 - deficiency; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Vimentin - analysis
• ISSN: 0065-1281; 1618-0372
• DOI: 10.1016/j.acthis.2012.02.006

Hyperthermia is used in cancer therapy, however much remains to be discovered regarding its mechanisms of action at the cellular level. In this study, the effects of hyperthermia on cell death, survival, morphology and the cytoskeleton were investigated in a non-small cell lung cancer cell line, H1299. Despite the fact that this cell line is widely used in research, it has not yet been tested for heat shock sensitivity. Cells were given a 30-min heat shock at 43.5°C and 45°C and left to recover at 37°C for 24 and 48h. 24h after heat shock treatment, we monitored changes in the organization of the cytoskeleton using immunofluorescence microscopy. The number of actin stress fibers was significantly reduced, microtubules formed a looser meshwork, a portion of the cells possessed multipolar mitotic spindles, whereas vimentin filaments collapsed into perinuclear complexes. 48h following heat stress, most of the cells showed recovery of the cytoskeleton, however we observed a considerable number of giant cells that were multinucleated or contained one enlarged nucleus. The data obtained by MTT assay showed a dose-dependent decrease of cell viability, while flow cytometric analysis revealed an increase in the number of cells with externalized phosphatidylserine. The results suggest that one of the modes of heat-induced cell death in H1299 cells is mitotic catastrophe, which probably ends in apoptosis.