Reduced joining of DNA ends correlates with chromosomal instability in three melanoma cell lines.
Chromosomal instability plays a pivotal role in multistep carcinogenesis by facilitating the acquisition of the multiple genetic alterations necessary for malignant transformation. In order to study the role of abnormal DNA repair in malignant melanoma, we measured the ability of cell lines from malignant melanoma and that of primary melanocytes to process 4 different kinds of DNA damage (pyrimidine dimers, oxidative DNA lesions, replication errors, and DNA double-strand breaks) using 4 different plasmid assays. Based on the number of chromosomes, the DNA index, and the rates of spontaneous micronuclei, the chromosomal stability in primary melanocytes and the melanoma line LIBR was characterized as being high, intermediate in M1, and low in MeWo. Repair of UVB photoproducts, of oxidative DNA damage, and of replication errors was not impaired in any melanoma line. Using linearized shuttle vector plasmid pZ189, LIBR cells and primary melanocytes exhibited a high efficiency of joining overlapping DNA ends, reflecting proficient repair of DNA double-strand breaks. Joining efficiency was reduced slightly in M1 and 2.9-fold in MeWo. This indicates that in the melanoma cell lines studied here, an increase in chromosomal instability is accompanied by a pronounced impairment in the ability to join DNA ends. Although a small sample was studied, this raises the possibility that defects in DNA end joining may also contribute to genetic instability and chromosome aberrations in melanoma.
Rünger
Thomas M
TM
Department of Dermatology, Boston University School of Medicine, Boston, MA 02118, USA. truenger@bu.edu
Kotas
Markus
M
Poot
Martin
M
Leverkus
Martin
M
Epe
Bernd
B
Jeggo
Penelope A
PA
Hellfritsch
Doris
D
D002869:Chromosome Aberrations; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D004273:DNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008545:Melanoma / Q000235:genetics*; D048629:Micronuclei, Chromosome-Defective; D009154:Mutation; D026082:Singlet Oxygen / Q000378:metabolism / Q000633:toxicity; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays / Q000009:adverse effects
D004273:DNA, Neoplasm; D026082:Singlet Oxygen