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DNA Repair and Repair Fidelity in Metastatic Variants of the B16 Murine Melanoma

Lookup NU author(s): Professor John LunecORCiD, Dr Gajanan Sherbet


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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 Bl 6 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.

Publication metadata

Author(s): Usmani BA, Lunec J, Sherbet GV

Publication type: Article

Publication status: Published

Journal: Journal of Cellular Biochemistry

Year: 1993

Volume: 51

Issue: 3

Pages: 336-344

Print publication date: 01/03/1993

ISSN (print): 0730-2312

ISSN (electronic): 1097-4644


DOI: 10.1002/jcb.240510313


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