Abstract
A series of eight oral epithelial cell lines derived from untreated human oral squamous cell carcinomas, which had arisen in patients with different tobacco histories, were examined for the presence of human papillomavirus (HPV) DNA, expression of stable p53 protein and p53 point mutation. Polymerase chain reaction (PCR)-based screening, but not Southern blot analysis, showed HPV-16 early region sequences to be present at low copy number (< 1 copy per cell) in two cell lines at early passage (3-5) in vitro (H400, T45), implying that only subpopulations of cells harboured viral DNA. HPV sequences were undetectable in cells at later passage (12-15), suggesting that viral sequences had been lost during growth in vitro, or that negative selection of HPV-containing cells had occurred. High levels of p53 were detected in the two HPV-positive cell lines and in three others (H103, H314, H357) by Western blotting, suggesting expression of mutant (stable) p53 molecules. A sixth cell line (H157) expressed a truncated p53. Sequence analysis of exons 2-11 of the p53 gene revealed missense mutations in six cell lines, one of which (H413) did not result in high levels of protein, and nonsense mutations in the remaining two cell lines (H157, H376). The results suggest that p53 mutation is a frequent genetic event in oral cancer. In addition, the expression of mutant p53 in oral cancer cells does not preclude a papillomaviral aetiology for these tumours. Analysis of p53 expression alone may result in underestimation of the frequency of p53 mutations in human cancers.(ABSTRACT TRUNCATED AT 250 WORDS)
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