Elsevier

Oral Oncology

Volume 38, Issue 4, June 2002, Pages 383-390
Oral Oncology

A novel, de novo germline TP53 mutation in a rare presentation of the Li-Fraumeni syndrome in the maxilla

https://doi.org/10.1016/S1368-8375(01)00078-1Get rights and content

Abstract

We undertook the genetic analysis of a classic Li-Fraumeni syndrome (LFS) family with clustering of primary tumours including two maxillary sarcomas, a rare LFS site of tumour occurrence. Our aim was to investigate the presence of a specific type of TP53 mutation that could be associated with this unusual predilection of site for cancer occurrence.

Mutational screening of the coding region of TP53 revealed an A>T transversion in codon 144 of exon 5 (CAG>CTG, Gln>Leu) in the germline of one of the three affected members, with loss of heterozygosity (LOH) in the tumour tissue. All other affected members were negative for germline or somatic TP53 mutations. TP53 immunohistochemistry was uninformative.

The mutation we report is a de novo constitutional TP53 mutation that has not been previously described in the literature. It could explain the more burdened phenotype of the affected patient (died at 21 months). Alternative mechanisms to explain the overall family phenotype are discussed.

Introduction

Cancer occurrence is principally sporadic; however, non-random clusters of tumours have been recognised to occur since the mid-nineteenth century, when Rokitansky and Virchow suggested the concept of constitutional predisposition for neoplasia [1]. The first report in English was by Warren in 1837, as a brief footnote on a familial cancer case [2]. Subsequently, Sir James Paget [3], [4] in England and Paul Broca in France [5] wrote and reported on cancer heredity, with Broca being the first to attempt statistical analysis on his cases and correlation with cancer incidence and patterns in the general population. Virtually every type of cancer has since been reported to occur in a familial form, with hereditary cancer syndromes accounting for a small but significant percentage. This percentage varies greatly amongst different types of cancer. Inherited mutations in the breast cancer genes BRCA1 and BRCA2 are responsible for approximately 7% of breast cancer and 10% of ovarian cancers, while 20–25% of all medullary thyroid carcinomas are due to inherited mutations in the RET (receptor protein tyrosine kinase) proto-oncogene [6].

Li-Fraumeni (LFS) syndrome, described in 1969 [7], [8], is a rare familial multicancer syndrome characterised by a striking aggregation of multiple tumours of early onset. It features five component tumours, namely sarcomas, breast cancer, adrenocortical carcinomas, leukaemias and brain tumours. A wide range of other tumours occasionally appears in the LFS phenotype. Studies on different family series confirmed that the familial clustering in LFS is due to inherited predisposition and not to environmental factors [9], [10], [11].

LFS is a highly penetrant autosomal dominant trait, with 40% reported risk for carriers to develop carcinoma (Ca) before the age of 16 [12], high mortality rates and increased frequency of second primaries [13]. Following systematic studies and collaboration between research groups, standard clinical criteria have been developed, and families are phenotypically classified as classic LFS (LFS-C) [14] and variant LFS (LFS-V) or LFS-like (LFL) [15], [16].

LFS was shown to be associated with germline TP53 mutations [17]. Although p53 is mutated in over 50% of sporadic human cancers, this is the only clinical condition described to date that features a high percentage of germline p53 mutations. However, in approximately 30% of classic LFS families no heritable p53 mutation can be detected after complete sequence of all exons including non-coding regions [18], [12], and TP53-negative families have been the subject of extensive research in the recent years.

Section snippets

Family history

The family we studied comprised of two adults and their three children. The pedigree featured five tumours (Fig. 1) in three of the five family members. The father (I.1, deceased) developed a maxillary chondrosarcoma at age 27. The second child (patient II.2, alive) developed a cluster of tumours that included an adrenocortical carcinoma with associated hermaphroditism at age 1, an osteosarcoma of the maxilla at age 26, and Bowen's disease at age 31. The son (II.3, deceased) died due to

DNA extraction and amplification

DNA extraction and purification was performed according to a modification of standard protocols [20], [21]. Purification by phenol-chloroform extraction was additionally performed on the paraffin tissue DNA [22].

PCR amplification followed. Sets of sense and antisense primers were used for the coding exons 2-11, including 10 base pairs of the intron sequences upstream and downstream of each exon, to include the splicing sites (Table 1) [23]. Exons 2 and 3 and 8 and 9 were amplified as single

Results

SSCP/HA screening indicated heteroduplex conformational shifts in the normal tissue of all affected members, indicative of a germline mutation located in exon 5 of the p53 gene. However, these were not of the same band pattern (Fig. 2, lanes 3, 5, 7). These conformations were consistently present in repeated screenings of exon 5 on products of separate PCR amplifications. Conformational shifts indicative of normal polymorphisms in exon 4 of patient I.2 and exon 6 of patients I.2, II.2 and the

Characteristics of the identified mutation

We identified a germline A>T transversion in codon 144, exon 5 of the TP53 gene in one of the three affected members (patient II.3). Analysis of the p53 and LFS mutational databases indicated that, at the germline level, the mutation we document has not been previously reported in the literature. To our knowledge, there are no reported germline mutational events of any type at position 144, rendering this a novel mutational site for Li-Fraumeni syndrome, even though identified in only one

Electronic database information

URLs for data in this article are as follows:

Institut Curie, http://perso.curie.fr/Thierry.Soussi/p53database.htm for somatic, germline and cell-line p53 mutation database

Charles University, Faculty of Medicine, Prague, http://www.lf2.cuni.cz/projects/germline_mut_p53.htm for LFS mutational database

UMD software, http://www.umd.necker.fr/ for position mutational analysis

EMBL database, http://www.ebi.ac.uk/cgi-bin/emblfetch for TP53 sequence

Acknowledgements

We would like to thank Professor Newell Johnson, Dr. Ann Gerling, Mr. Roger Rees, Dr. S. Sousha and Dr. Evan Reed for provision of clinical information and histology material, as well as Dr. Merrill Griffiths for her kind review of histopathogical material. Special thanks to Drs. Frederick Li and Joseph Fraumeni, Dr. Thierry Soussi and Dr. Diana Barnes for their comments and advice.

References (59)

  • J. Paget

    Lectures on Surgical Pathology

    (1853)
  • J. Paget

    Med. Times Gaz.

    (1857)
  • P. Broca

    Traite des Tumeurs

    (1866)
  • T.S. Frank

    Hereditary cancer syndromes

    Arch. Pathol. Lab. Med.

    (2001)
  • F.P. Li et al.

    Rhabodmyosarcoma in children; epidemiologic study and identification of a cancer family syndrome

    J. Natl. Cancer Inst.

    (1969)
  • F.P. Li et al.

    Soft-tissue sarcomas, breast cancer and other neoplasms. A familial syndrome?

    Ann. Intern. Med.

    (1969)
  • J.M. Birch et al.

    Excess risk of breast cancer in the mothers of children with soft-tissue sarcomas

    Br. J. Cancer

    (1984)
  • J.M. Birch et al.

    Cancer in the families of children with soft-tissue sarcomas

    Cancer

    (1990)
  • L.C. Strong et al.

    Cancer in survivors of childhood soft tissue sarcoma and their relatives

    J. Natl. Cancer Inst.

    (1987)
  • C. Eng et al.

    Third international workshop on collaborative interdisciplinary studies of p53 and other predisposing genes in Li-Fraumeni syndrome

    Cancer Epidemiol., Biomarkers and Prev.

    (1997)
  • F.P. Li et al.

    Prospective study of a cancer family syndrome

    JAMA

    (1982)
  • F.P. Li et al.

    A cancer family syndrome in twenty-four kindreds

    Cancer Res.

    (1988)
  • J.M. Birch et al.

    Prevalence and diversity of constitutional mutations in the p53 gene among 21 Li-Fraumeni families

    Cancer Res.

    (1994)
  • R.A. Eeles

    Germline mutations in the TP53 gene

    Cancer Surv.

    (1995)
  • D. Malkin et al.

    Germline p53 mutations in a familial syndrome of breast cancer, sarcomas and other neoplasms

    Science

    (1990)
  • J.M. Varley et al.

    Germline mutations of TP53 gene in Li-Fraumeni families: an extended study of 39 families

    Cancer Res.

    (1997)
  • D.K. Lahiri et al.

    A rapid non-enzymatic method for the preparation of HMW DNA from blood for RFLP studies

    Nucl. Acids Res.

    (1991)
  • Z. Onadim et al.

    Application of PCR amplification of DNA from paraffin embedded tissue sections to linkage analysis in familial retinoblastoma

    J. Med. Genet.

    (1991)
  • Maniatis T, Fritschef, Sambrook J. Molecular cloning: a laboratory manual, Vol 2. Cold Spring Harbour Press, New York,...
  • Cited by (0)

    View full text