DNA repair and repair fidelity in metastatic variants of the B16 murine melanoma.
We have examined the concept of genomic instability in relation to the metastatic progression of low (F1) and high metastasis (BL6, ML8) clones of the B16 mouse melanoma, by using a mutation assay, and DNA strand break repair and repair fidelity assays. The frequency of induced ouabain resistant colonies between the variant cell lines was consistent with the difference between their metastatic properties. Survival data for X-irradiation and bleomycin were similar among the 3 cell lines. When X-rays or bleomycin were used to induce strand breakage, no difference was detectable in either the rate or extent of DNA repair using the techniques of alkaline unwinding and alkaline elution for total strand breaks, and neutral elution for double strand breaks. DNA repair fidelity was measured using the PMH16 plasmid. A Kpn I restriction site was used to introduce a break within the gpt gene of the plasmid, prior to transfection. We found that approximately 100% and approximately 65% of the highly metastatic ML8 and Bl6 clones, respectively, religated the gene with the required fidelity, compared with only approximately 25% of the low metastasis F1 clones. In summary, the metastatic variants show similar sensitivities to X-irradiation and bleomycin, but a differential response to EMS. This difference is not reflected in any subsequent DNA strand break religation, but the variants do differ in their fidelity of repair. However, although the fidelity of DNA religation is related to metastatic potential, it is not consistent with the mutation frequency data.
Usmani
B A
BA
Cancer Research Unit, University of Newcastle upon Tyne, Medical School, United Kingdom.
Lunec
J
J
Sherbet
G V
GV
D000818:Animals; D001761:Bleomycin / Q000633:toxicity; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D004260:DNA Repair; D005020:Ethyl Methanesulfonate / Q000633:toxicity; D008545:Melanoma / Q000235:genetics* / Q000556:secondary; D051379:Mice; D008957:Models, Genetic; D009154:Mutation; D015203:Reproducibility of Results; D014162:Transfection; D014407:Tumor Cells, Cultured
D001761:Bleomycin; D005020:Ethyl Methanesulfonate