Analysis for loss of heterozygosity (LOH) of p53 allele in tumors derived from p53+/- and CD-1 mice following repeated subcutaneous injections of solutions containing antioxidants.
Genomic DNA was isolated from subcutaneous masses observed in CD-1 and p53+/- heterozygous mice during the course of carcinogenicity studies in the vehicle control groups. These masses resulted after daily subcutaneous injection of an antioxidant vehicle with a pH adjusted to 3-4. The vehicle was 1.0% ascorbic acid plus 0.05% sodium metabisulfite in 0.75% saline in a dosing volume of 10 ml/kg/day. These masses were first palpable after 13 and 37 weeks of dosing among p53+/- and CD-1 mice, respectively. By week 26, the incidence of these masses was 89% and 80% in male and female p53+/- mice, respectively (n = 15 mice/sex) and was 0% in both male and female CD-1 mice (n = 60 mice/sex). These masses originated from panniculus carnosus muscle. Histopathological examination of the p53+/- mouse masses indicated the tumors to be sarcomas of spindle-cell origin. The histopathological examination of the masses in the CD-1 mice revealed fibrosarcomas. Five mice/sex/strain were randomly selected from a pool of mice that developed these masses in the course of the two studies. Frozen tissues from these masses were used to examine the DNA for loss of the functional p53 allele in the p53+/- mice (i.e., loss of heterozygosity, or LOH) or for loss of one of the alleles in the wild type (p53+/+) CD-1 mice by the polymerase chain reaction (PCR) technique. Loss of the functional allele was observed only in the tumor from one p53+/- male mouse. These results support a nongenotoxic mechanism for these injection site masses.
Youssef
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AF
TAP Pharmaceutical Products Inc, Lake Forest, IL 60045, USA. ashraf.youssef@tap.com
Borellini
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F
Jacobson-Kram
D
D
Fort
F L
FL
D000483:Alleles; D000818:Animals; D000975:Antioxidants / Q000008:administration & dosage / Q000633:toxicity; D001205:Ascorbic Acid / Q000008:administration & dosage / Q000633:toxicity*; D015197:Carcinogenicity Tests; D005260:Female; D005354:Fibrosarcoma / Q000139:chemically induced* / Q000473:pathology; D006579:Heterozygote; D007279:Injections, Subcutaneous; D019656:Loss of Heterozygosity / Q000187:drug effects* / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D012208:Rhabdomyosarcoma / Q000139:chemically induced* / Q000473:pathology; D012680:Sensitivity and Specificity; D012983:Soft Tissue Neoplasms / Q000139:chemically induced* / Q000473:pathology; D013447:Sulfites / Q000008:administration & dosage / Q000633:toxicity*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*
D000975:Antioxidants; D013447:Sulfites; D016159:Tumor Suppressor Protein p53; C005200:sodium metabisulfite; D001205:Ascorbic Acid