Presence of human papillomavirus sequences in tumour-derived human oral keratinocytes expressing mutant p53

William Andrew Yeudall, I. C. Paterson, V. Patel, S. S. Prime

Research output: Contribution to journalArticle

50 Citations (Scopus)

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. In contrast to other studies, we demonstrate that positive staining of p53 in oral cancer does not necessarily reflect a tobacco aetiology.

Original languageEnglish (US)
Pages (from-to)136-143
Number of pages8
JournalEuropean Journal of Cancer. Part B: Oral Oncology
Volume31
Issue number2
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Keratinocytes
Cell Line
Mouth Neoplasms
Neoplasms
Tobacco
Human papillomavirus 16
Nonsense Codon
p53 Genes
Viral DNA
Mutation Rate
Missense Mutation
Southern Blotting
Point Mutation
Sequence Analysis
Squamous Cell Carcinoma
Exons
Proteins
Western Blotting
Epithelial Cells
Staining and Labeling

Keywords

  • chemical carcinogens
  • gene mutation
  • human papillomavirus
  • oral cancer
  • p53
  • tumour progression
  • tumour suppressor
  • viral oncogenesis

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Presence of human papillomavirus sequences in tumour-derived human oral keratinocytes expressing mutant p53. / Yeudall, William Andrew; Paterson, I. C.; Patel, V.; Prime, S. S.

In: European Journal of Cancer. Part B: Oral Oncology, Vol. 31, No. 2, 01.01.1995, p. 136-143.

Research output: Contribution to journalArticle

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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. In contrast to other studies, we demonstrate that positive staining of p53 in oral cancer does not necessarily reflect a tobacco aetiology.",
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