Patches of mutant p53-immunoreactive epidermal cells induced by chronic UVB Irradiation harbor the same p53 mutations as squamous cell carcinomas in the skin of hairless SKH-1 mice.
Treatment of SKH-1 hairless mice with UVB (30 mJ/cm(2)) twice a week for 20 weeks results in the formation of cellular patches, long before the appearance of tumors, that are visualized in epidermal sheets with an antibody (PAb240) recognizing mutated p53 protein. Direct sequencing analysis of the whole coding region of the p53 gene (exons 2-11) detected one or two mutations in 64.4% of 104 analyzed patches and no mutations in nonstained adjacent normal controls. Homozygous mutation was detected in 22.4% of the mutant patches. Except for two nonsense mutations, all others were missense (exons 4-9) and mostly (95.5%) at the DNA-binding domain. Primer extension analysis of cloned PCR fragments found three of four double-mutated patches harboring different mutations in separate alleles. All mutation hotspots reported earlier in UVB-induced mouse squamous cell carcinomas (SCC) at codons 270 (Arg --> Cys), 149 (Pro --> Ser), 275 (Pro --> Leu and Pro --> Ser), and 176 (His --> Tyr) with a frequency of 32.1%, 7.1%, 14.7%, and 3.2% were detected in epidermal patches at a frequency 47.7%, 9.1%, 4.5%, and 2.3%, respectively. Mutations at codons 210 and 191 found in patches at respective frequencies of 8.0% and 4.5% were not previously detected in UVB-induced mouse SCC. In summary, (a) the p53 mutation profile of UVB-induced skin patches and SCC was very similar suggesting that patches are precursor lesions for SCC, (b) a small number of patches harbored mutations that were not before observed in SCC from UVB-treated mice, and (c) about 36% of the patches did not harbor a p53 mutation.
Kramata
Pavel
P
Susan Lehman Cullman Laboratory for Cancer Research, Department of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854-8020, USA. kramata@rci.rutgers.edu
Lu
Yao-Ping
YP
Lou
You-Rong
YR
Singh
Rayvita N
RN
Kwon
Sherry M
SM
Conney
Allan H
AH
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000209:etiology / Q000235:genetics*; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000528:radiation effects; D003062:Codon; D005260:Female; D016158:Genes, p53 / Q000528:radiation effects*; D015603:Keratinocytes / Q000528:radiation effects; D051379:Mice; D008812:Mice, Hairless; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009381:Neoplasms, Radiation-Induced / Q000209:etiology / Q000235:genetics*; D012867:Skin / Q000166:cytology / Q000473:pathology / Q000528:radiation effects*; D012878:Skin Neoplasms / Q000209:etiology / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000528:radiation effects; D014466:Ultraviolet Rays / Q000009:adverse effects
D003062:Codon; D016159:Tumor Suppressor Protein p53