Cytoskeletal and nuclear alterations in human lung tumor cells: a confocal microscope study

Tumor cells generally present various types of nuclear alterations, which can be associated with genetic instability. The origin and mechanism of formation of the nuclear alterations are largely unknown, with the micronucleus being the most well studied alteration. The purpose of this study was to characterize the cytoskeleton filaments and to analyze the possible association between nuclear alterations and the cytoskeleton in the human lung carcinoma cells HK2 and A549. The cytoskeleton analysis was performed by using antibodies against lamin B, vimentin, cytokeratin-8, and alpha-tubulin and the secondary antibody labeled with FITC. The analysis of the actin filament was made with phalloidin-TRITC. The analyses of cytoskeleton were performed from optical sections obtained by confocal laser scanning microscopy. Filaments of the cytoskeleton of tumor cells present some differences in their distribution pattern and their expression when compared with the filaments of normal cells. The HK2 cells presented actin fibers arranged either concentrically or in clusters and tubulin filaments arranged radially, while in the A549 cells the distribution pattern was similar to that of normal cells. The lamin B filaments were the most important to identify nuclear alterations. These alterations in cytoskeleton distribution could not be associated with nuclear alterations.