p53 gene and cell cycling in uveal melanoma.
OBJECTIVE
To determine whether alterations of p53, a tumor suppressor gene, were present in uveal melanoma, and to characterize further the nature of those changes.


METHODS
Immunohistochemical analysis with a monoclonal antibody was used to determine whether alterations of p53 were present in 35 enucleated archival uveal melanomas. Further characterization was done by comparing the p53 gene and cell cycling status by using bromodeoxyuridine staining. The alterations in p53 were characterized using polymerase chain reaction single-strand conformational polymorphism analysis and sequencing.


RESULTS
Four of 35 uveal melanomas showed low levels (0.5% to 5.0%) of positive immunostaining for altered p53 in tumor cell nuclei using monoclonal antibody DO-7. These four tumors had the three highest and the 14th highest bromodeoxyuridine labeling indices, ranging from 1.3% to 7.0%. Polymerase chain reaction single-strand conformational polymorphism analysis of p53 exons 5 to 8 was performed on three p53-positive and six p53-negative tumors, and no altered motility shifts were detected. Sequencing of one of the positive staining specimens confirmed no mutations in exons 5 through 8 in the p53 gene. Double immunohistochemical labeling for both bromodeoxyuridine and p53 in one tumor showed that most of p53-positive cells were in S phase.


CONCLUSIONS
Mutation of p53 is an uncommon event in uveal melanomas. Nuclear accumulation of p53 protein was found in three of the four tumors with the highest levels of cell cycling.
Kishore
K
K
Department of Ophthalmology, University of California, San Francisco, CA 94143, USA.
Ghazvini
S
S
Char
D H
DH
Kroll
S
S
Selle
J
J
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D001973:Bromodeoxyuridine; D002453:Cell Cycle / Q000235:genetics; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000737:chemistry; D004273:DNA, Neoplasm / Q000032:analysis; D005260:Female; D005500:Follow-Up Studies; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008545:Melanoma / Q000737:chemistry / Q000235:genetics*; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D016159:Tumor Suppressor Protein p53 / Q000032:analysis; D014604:Uveal Neoplasms / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D004273:DNA, Neoplasm; D016159:Tumor Suppressor Protein p53; D001973:Bromodeoxyuridine