Isolation of V79 fibroblast cell lines containing elevated metallothionein levels that have increased resistance to the cytotoxic effects of ultraviolet-A radiation.
Isolated clones of V79 Chinese hamster lung fibroblasts, selected for resistance against cadmium toxicity, were exposed to monochromatic 365 nm ultraviolet-A (UVA; 320 nm to visible light) radiation and examined for cell survival. All three of the Cd-resistant V79 clones (V79Cd) tested exhibited significant increases in survival after irradiation compared with control cultures similar to the increased survival observed in Zn acetate-induced V79 cells. Dose-modifying factors calculated for these survival experiments were all approximately 1.5. When characterized for steady-state levels of metallothionein (MT) mRNA and associated Cd-binding activity, all of the Cd-resistant V79Cd clones demonstrated elevated constitutive levels of both, implicating MT as the mechanism responsible for the observed cellular resistance to Cd and also to 365 nm UVA radiation. However, whereas levels of intracellular MT protein correlated with differences in survival against Cd, MT intracellular levels did not correlate well with protection against 365 nm UVA. Increased cell survival after exposure to 365 nm UVA radiation mediated by MT appeared to reach a threshold level and MT only provided a limited degree of protection. Since UVA radiation is known to cause cell death mediated through the intracellular generation of reactive oxygen species (ROS), these results suggest that the role of MT in ameliorating cellular photooxidative damage produced by UVA is by reducing intracellular ROS.
Dudek
E J
EJ
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, IL 60439-4833.
Peak
J G
JG
Roth
R M
RM
Peak
M J
MJ
D000818:Animals; D002104:Cadmium / Q000378:metabolism / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004351:Drug Resistance; D005347:Fibroblasts; D014644:Genetic Variation; D008168:Lung / Q000166:cytology; D008668:Metallothionein / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D011836:Radiation Tolerance / Q000502:physiology*; D012641:Selection, Genetic; D014466:Ultraviolet Rays / Q000009:adverse effects*
D012333:RNA, Messenger; D002104:Cadmium; D008668:Metallothionein