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Title: In vitro sensitivity of normal and hereditary retinoblastoma fibroblasts to DNA-damaging agents

Abstract

We investigated the ability of nine fibroblast cell strains from patients with the hereditary form of retinoblastoma (RB) to handle various types of DNA-damaging agents and compared the results with those obtained in nine normal strains. Cell strains were exposed to gamma-radiation, which causes DNA scission; actinomycin D, a DNA-intercalating agent; and mitomycin C, a bifunctional alkylating agent leading to DNA-DNA cross-linking. Cell strains were studied for their ability to survive in a cytotoxicity assay. Nine normal strains exhibited a mean D0 (inverse of the slope of the straight line portion of the survival curve) of 134-178 cGy after radiation exposure, compared to a range of 119-186 cGy in the nine RB strains (P = 0.33). Similarly, exposure to actinomycin D led to D0 values of 0.024-0.069 microgram/ml in the nine normal strains and D0 values of 0.016-0.067 microgram/ml in the RB strains (P = 0.64). The nine RB strains did exhibit a small overall increase in sensitivity after exposure to mitomycin C, with D0 values ranging from 0.14-0.32 microgram/ml versus 0.19-0.66 microgram/ml in the nine normal strains (P = 0.002); however, when the two most resistant normal strains were excluded from analysis, results were similar. Three RB cell strainsmore » derived from individuals who had either developed second cancers or who had a family history of additional sarcomas consistently exhibited increases in sensitivity to all three DNA-damaging agents studied compared with other hereditary RB cell strains as well as normal strains. The results suggest that normal human fibroblast cell strains exhibit a wide response to DNA-damaging agents, especially chemical agents. Most hereditary RB strains exhibit sensitivity well within the normal range; however, strains from RB patients predisposed to second cancers exhibit increases in sensitivity to DNA-damaging agents.« less

Authors:
;
Publication Date:
Research Org.:
Univ. of Minnesota Hospitals, Minneapolis
OSTI Identifier:
6928453
Alternate Identifier(s):
OSTI ID: 6928453
Resource Type:
Journal Article
Resource Relation:
Journal Name: Cancer Res.; (United States)
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; DNA; DAMAGE; FIBROBLASTS; SENSITIVITY; ACTINOMYCIN; ALKYLATING AGENTS; DNA REPAIR; GAMMA RADIATION; GLIOMAS; IN VITRO; MITOMYCIN; NEOPLASMS; PATIENTS; RADIOSENSITIVITY; RETINA; ANIMAL CELLS; ANTI-INFECTIVE AGENTS; ANTIBIOTICS; ANTIMITOTIC DRUGS; ANTINEOPLASTIC DRUGS; BIOLOGICAL RECOVERY; BIOLOGICAL REPAIR; BODY; BODY AREAS; CONNECTIVE TISSUE CELLS; DISEASES; DRUGS; ELECTROMAGNETIC RADIATION; EYES; FACE; HEAD; IONIZING RADIATIONS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANS; RADIATIONS; RECOVERY; REPAIR; SENSE ORGANS; SOMATIC CELLS 560120* -- Radiation Effects on Biochemicals, Cells, & Tissue Culture; 560300 -- Chemicals Metabolism & Toxicology

Citation Formats

Woods, W.G., and Byrne, T.D. In vitro sensitivity of normal and hereditary retinoblastoma fibroblasts to DNA-damaging agents. United States: N. p., 1986. Web.
Woods, W.G., & Byrne, T.D. In vitro sensitivity of normal and hereditary retinoblastoma fibroblasts to DNA-damaging agents. United States.
Woods, W.G., and Byrne, T.D. Mon . "In vitro sensitivity of normal and hereditary retinoblastoma fibroblasts to DNA-damaging agents". United States. doi:.
@article{osti_6928453,
title = {In vitro sensitivity of normal and hereditary retinoblastoma fibroblasts to DNA-damaging agents},
author = {Woods, W.G. and Byrne, T.D.},
abstractNote = {We investigated the ability of nine fibroblast cell strains from patients with the hereditary form of retinoblastoma (RB) to handle various types of DNA-damaging agents and compared the results with those obtained in nine normal strains. Cell strains were exposed to gamma-radiation, which causes DNA scission; actinomycin D, a DNA-intercalating agent; and mitomycin C, a bifunctional alkylating agent leading to DNA-DNA cross-linking. Cell strains were studied for their ability to survive in a cytotoxicity assay. Nine normal strains exhibited a mean D0 (inverse of the slope of the straight line portion of the survival curve) of 134-178 cGy after radiation exposure, compared to a range of 119-186 cGy in the nine RB strains (P = 0.33). Similarly, exposure to actinomycin D led to D0 values of 0.024-0.069 microgram/ml in the nine normal strains and D0 values of 0.016-0.067 microgram/ml in the RB strains (P = 0.64). The nine RB strains did exhibit a small overall increase in sensitivity after exposure to mitomycin C, with D0 values ranging from 0.14-0.32 microgram/ml versus 0.19-0.66 microgram/ml in the nine normal strains (P = 0.002); however, when the two most resistant normal strains were excluded from analysis, results were similar. Three RB cell strains derived from individuals who had either developed second cancers or who had a family history of additional sarcomas consistently exhibited increases in sensitivity to all three DNA-damaging agents studied compared with other hereditary RB cell strains as well as normal strains. The results suggest that normal human fibroblast cell strains exhibit a wide response to DNA-damaging agents, especially chemical agents. Most hereditary RB strains exhibit sensitivity well within the normal range; however, strains from RB patients predisposed to second cancers exhibit increases in sensitivity to DNA-damaging agents.},
doi = {},
journal = {Cancer Res.; (United States)},
number = ,
volume = ,
place = {United States},
year = {Mon Dec 01 00:00:00 EST 1986},
month = {Mon Dec 01 00:00:00 EST 1986}
}
  • Neurofibromatosis (NF) is an autosomal dominant disorder associated with various constitutional abnormalities as well as a striking predisposition for malignant and nonmalignant neoplasms, both in cells originating in and not originating in the neural crest. We have examined the sensitivity of cultured skin fibroblasts from patients with neurofibromatosis to several types of DNA damage. Fibroblasts in Dulbecco's modified Eagle's medium were plated at 10(2) to 2 X 10(4) cells per 75 cm2 tissue culture plates, and exposed to various doses of gamma radiation (leads to DNA scission), actinomycin D, or mitomycin C. Cells were reincubated for 15 to 40 daysmore » until surviving colonies exhibited greater than 30-50 cells. Plates were then stained with 1% methylene blue and the colonies counted, with surviving fraction determined relative to plating efficiency. Nine skin fibroblast cell strains from normal individuals were studied as controls. One neurofibromatosis (NF) cell strain, SB23, exhibited normal sensitivity to all three DNA-damaging agents studied in early (7-8) and middle (12-13) in vitro passage. Strain GM0622, on the other hand, exhibited normal sensitivity to the three DNA-damaging agents studied at early passage, but showed a significant decrease in survival after exposure to both gamma radiation (D0 = 106 rad) and actinomycin D (D0 = 0.024 mcg/ml) with increasing passage. Strain GM1639 exhibited decreased survival after actinomycin D exposure at early passage (D0 = 0.017 mcg/ml), with normal survival after exposure to gamma radiation and mitomycin C at the same passage.« less
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  • Mutation in the bimD gene of Aspergillus nidulans results in a mitotic block in anaphase characterized by a defective mitosis. Mutation in bimD also confers, at temperatures permissive for the mitotic arrest phenotype, an increased sensitivity to DNA damaging agents, including methyl methanesulfonate and ultraviolet light. In order to better understand the relationship between DNA damage and mitotic progression, the authors cloned the bimD gene from Aspergillus. A cosmid containing the bimD gene was identified among pools of cosmids by cotransformation with the nutritional selective pyrG gene of a strain carrying the recessive, temperature-sensitive lethal bimD6 mutation. The bimD genemore » encodes a predicted polypeptide of 166,000 daltons in mass and contains amino acid sequence motifs similar to those found in some DNA-binding transcription factors. These sequences include a basic domain followed by a leucine zipper, which together are called a bZIP motif, and a carboxyl-terminal domain enriched in acidic amino acids. Overexpression of the wild-type bimD protein resulted in an arrest of the nuclear division cycle that was reversible and determined to be in either the G[sub 1] or S phase of the cell cycle. The data suggest that bimD may play an essential regulatory role relating to DNA metabolism which is required for a successful mitosis. 7l refs., 7 figs., 1 tab.« less
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