DNA repair in lymphoblastoid cell lines established from human genetic disorders |
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Authors: | EE Henderson Rene Ribecky |
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Institution: | 1. Department of Microbiology and Immunology, Temple University School of Medicine, Temple University Life Sciences Center, Philadelphia, PA 19140 U.S.A.;2. Fels Research Institute, Temple University School of Medicine, Temple University Life Sciences Center, Philadelphia, PA 19140 U.S.A. |
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Abstract: | Lymphoblastoid cell lines (LCLs) established from chromosomal breakage syndromes or related genetic disorders have been used to study the effects of mutagens on human lymphoid cells. The disorders studied include xeroderma pigmentosum, ataxia telangiectasia, Fanconi's anemia, Bloom's syndrome and Cockayne's syndrome. Three approaches were used to assess the cells' ability to cope with a particular mutagen: (1) assaying recovery of DNA snythetic capabilities as measured by 3H]thymidine (dT) incorporation; (2) measurements of classical excision DNA repair by isopyknic sedimentation of DNA density labeled with 5-bromo-2-deoxyuridine (BrdU); (3) determining cell survival by colony formation in microtiter plates. LCLs established from xeroderma pigmentosum showed increased sensitivities to ultraviolet (354 nm) light and N-acetoxy-2-acetylaminofluorene (AAAF) as determined by DNA synthesis or colony formation and had diminished levels of excision-repair. Cockayne's syndrome LCLs, on the other hand, had increased sensitivities to ultraviolet (UV) light, AAAF and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) while showing near normal levels of DNA-repair after treatment with each agent. An LCL established from ataxia telangiectasia had decreased DNA repair synthesis and defective colony-forming ability following treatment with MNNG. LCLs, in addition to ease of establishment, appear likely to provide useful material for the study of DNA repair replication and its relationship to carcinogenesis. |
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Keywords: | AAAF BrdU 5-bromo-2-deoxyuridine dT thymidine EBV Epstein-Barr virus LCL lymphoblastoid cell line MNNG SSC 0 15 M NaCl plus 0 015 M sodium citrate |
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