Low frequency of codon 61 Ha-ras mutations and lack of keratin 13 expression in 7,12-dimethylbenz[a]-anthracene-induced hamster skin tumors.
Alterations in the pattern of keratin expression are a common feature of skin-tumor development. In this study, we investigated whether the loss of epidermal keratin 1 (K1) and its replacement by mucosal keratin 13 (K13) is unique to mouse skin tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA), since it has been reported that human epidermal tumors do not exhibit aberrant expression of K13. With that purpose, we analyzed the keratin profiles of 16 DMBA-induced hamster skin tumors using monospecific antibodies against K1 and K13. Although all the tumors expressed K1, they also showed an overall tendency towards loss of this keratin; furthermore, none of the tumors expressed K13. Previous studies have suggested that the induction of K13 in mouse skin is related to the mutation of the Ha-ras gene by the initiating agent DMBA, a mutation consistently found in murine DMBA/TPA-induced tumors and rarely found in human skin tumors. Therefore, we also evaluated the tumors for the presence of codon-61 mutations by direct sequencing of DNA extracted from paraffin-embedded tissue sections. Only three tumors showed an A-->T transversion in the second nucleotide of Ha-ras codon 61. However, presence of the mutation did not correlate with K1 staining. Although hamster skin tumors were induced by the same initiator as were mouse skin tumors, hamster skin tumors did not show the same keratin profile. Moreover, their immunohistochemical expression of K1 and K13 and their codon 61 sequences resembled that of their human counterparts. These results suggest that the aberrant expression of K13 may be unique to murine skin. Furthermore, although codon 61 Ha-ras mutation appears to be related to keratin alterations in the mouse model, this mutation is not sufficient to produce the same biochemical changes in other species.
Robles
A I
AI
University of Texas M. D. Anderson Cancer Center, Science Park, Research Division, Smithville 78957.
Gimenez-Conti
I B
IB
Roop
D
D
Slaga
T J
TJ
Conti
C J
CJ
D015127:9,10-Dimethyl-1,2-benzanthracene; D000818:Animals; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000139:chemically induced / Q000737:chemistry / Q000235:genetics*; D002610:Cheek; D006224:Cricetinae; D004273:DNA, Neoplasm / Q000032:analysis; D011905:Genes, ras / Q000187:drug effects*; D007124:Immunoenzyme Techniques; D007633:Keratins / Q000032:analysis*; D008297:Male; D008647:Mesocricetus; D008969:Molecular Sequence Data; D010212:Papilloma / Q000139:chemically induced / Q000737:chemistry / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012878:Skin Neoplasms / Q000139:chemically induced / Q000737:chemistry* / Q000235:genetics*; D013045:Species Specificity
D004273:DNA, Neoplasm; D015127:9,10-Dimethyl-1,2-benzanthracene; D007633:Keratins