Resistance to DNA denaturation in irradiated Chinese hamster V79 fibroblasts is linked to cell shape.
Exponentially growing Chinese hamster V79-171b lung fibroblasts seeded at high density on plastic (approximately 7 x 10(3) cells/cm2) flatten, elongate, and produce significant amounts of extracellular fibronectin. When lysed in weak alkali/high salt, the rate of DNA denaturation following exposure to ionizing radiation is exponential. Conversely, cells plated at low density (approximately 7 x 10(2) cells/cm2) on plastic are more rounded 24 h later, produce little extracellular fibronectin, and display unusual DNA denaturation kinetics after X-irradiation. DNA in these cells resists denaturation, as though "constraints" to DNA unwinding have developed. Cell doubling time and distribution of cells in the growth cycle are identical for both high and low density cultures as is cell survival in response to radiation damage. The connection between DNA conformation and cell shape was examined further in low density cultures grown in conditioned medium. Under these conditions, cells at low density were able to elongate, and DNA denaturation of low density cultures was identical to that of high density cultures. Conversely, cytochalasin D, which interferes with actin polymerization causing cells to "round up" and release fibronectin, allowed development of constraints in high density cultures. These results suggest that DNA conformation is sensitive to changes in cell shape which result when cells are grown in different environments. However, these changes in DNA conformation detected by the DNA unwinding assay do not appear to play a direct role in radiation-induced cell killing.
Olive
P L
PL
British Columbia Cancer Research Centre, Vancouver, Canada.
Vanderbyl
S
S
MacPhail
S H
SH
D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000528:radiation effects; D002478:Cells, Cultured / Q000166:cytology / Q000528:radiation effects*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000528:radiation effects*; D004249:DNA Damage; D005353:Fibronectins / Q000502:physiology; D008168:Lung / Q000166:cytology; D009691:Nucleic Acid Denaturation
D005353:Fibronectins; D004247:DNA