TITLE:
Changes in the Mechanical Environment of the Nucleus with Cell Crowding and Its Effects on DNA Damage Resistance
AUTHORS:
Kazuaki Nagayama, Yuto Hirooka
KEYWORDS:
Cell Biomechanics, Mechanobiology, Mechanical Properties, DNA Damage, γ-H2A.X
JOURNAL NAME:
Journal of Biosciences and Medicines,
Vol.13 No.1,
January
21,
2025
ABSTRACT: Nuclear DNA, which is essential for the transmission of genetic information, is constantly damaged by external stresses and is subsequently repaired by the removal of the damaged region, followed by resynthesis of the excised region. Accumulation of DNA damage with failure of repair processes leads to fatal diseases such as cancer. Recent studies have suggested that intra- and extra-nuclear environments play essential roles in DNA damage. However, numerous questions regarding the role of the nuclear mechanical environment in DNA damage remain unanswered. In this study, we investigated the effects of cell confluency (cell crowding) on the morphology of cell nuclei, and cytoskeletal structures, and DNA damage in NIH3T3 skin fibroblasts and HeLa cervical cancer cells. Although nuclear downsizing was observed in both NIH3T3 and HeLa cells with cell crowding, intracellular mechanical changes in the two cell types displayed opposite tendencies. Cell crowding in NIH3T3 cells induced reinforcement of actin filament structures, cell stiffening, and nuclear downsizing, resulting in a significant decrease in endogenous DNA damage, whereas cell crowding in HeLa cells caused partial depolymerization of actin filaments and cell softening, inducing endogenous DNA damage. Ultraviolet (UV) radiation significantly increased DNA damage in NIH3T3; however, this response did not change with cell crowding. In contrast, UV radiation did not cause DNA damage in HeLa cells under either sparse or confluent conditions. These results suggested that cell crowding significantly influenced endogenous DNA damage in cells and was quite different in NIH3T3 and HeLa cells. However, cell crowding did not affect the UV-induced DNA damage in either cell type.