Germline TP53 mutations in BRCA1 and BRCA2 mutation-negative French Canadian breast cancer families

EPIDEMIOLOGY

Germline TP53 mutations in BRCA1 and BRCA2 mutation-negative French Canadian breast cancer families

Suzanna L. Arcand Æ Christine M. Maugard Æ Parviz Ghadirian ÆAndre Robidoux Æ Chantal Perret Æ Phil Zhang Æ Eve Fafard ÆAnne-Marie Mes-Masson Æ William D. Foulkes Æ Diane Provencher ÆSteven A. Narod Æ Patricia N. Tonin

Received: 20 April 2007 / Accepted: 23 April 2007

� Springer Science+Business Media B.V. 2007

Abstract About 40% of French Canadian breast and/or

ovarian cancer families harbor germline BRCA1 or

BRCA1 mutations where common mutations account for

about 84% of all mutations identified in cancer families.

Within a series of BRCA1 and BRCA2 mutation-negative

families, a germline TP53 13398 G>A (Arg213Gln)

mutation was identified, which was selected for mutation

analysis in this gene because of a family history consistent

with Li–Fraumeni syndrome (LFS). Given the founder ef-

fects in this population, the 13398 G>A mutation was

screened in series of 52 BRCA1 and BRCA2 mutation-

negative cancer families, and a mutation-positive family

was identified. However, pedigree inspection and

expansion of mutation-positive families with the same

mutation revealed that they were closely related to each

other. To further characterize the contribution of TP53 in

cancer families, mutation analysis was performed in the

remaining BRCA1 and BRCA2 mutation-negative cancer

families. Thirty sequence variants were identified, the

majority of which occur in intronic sequences and are not

predicted to affect the functionality of TP53. However, the

14538 G>A (Arg290His) mutation was identified in a

family which did not exhibit features consistent with LFS

or Li–Fraumeni-like (LFL) syndrome. Neither of the TP53

mutations was detected in 381 French Canadian women

with breast cancer diagnosed before 50 years of age not

S. L. Arcand � W. D. Foulkes � D. Provencher �P. N. Tonin

The Research Institute of the McGill University Health Centre,

Montreal, QC, Canada

C. M. Maugard

Service de Medecine Genique, Centre Hospitalier de l’Universite

de Montreal (CHUM), Montreal, QC, Canada

C. M. Maugard � A.-M. Mes-Masson

Departement de medecine, Universite de Montreal, Montreal,

QC, Canada

P. Ghadirian � E. Fafard

Epidemiology Research Unit, Research Centre, CHUM—Hotel-

Dieu, Montreal, QC, Canada

A. Robidoux

Department of Surgery, CHUM—Hotel-Dieu, Montreal, QC,

Canada

C. Perret � A.-M. Mes-Masson � D. Provencher

Centre de Recherche du Centre Hospitalier de l’Universite de

Montreal/Institut du Cancer de Montreal, Hopital Notre-Dame,

Montreal, QC, Canada

P. Zhang � S. A. Narod

Women’s College Research Institute, Women’s College Hospital

and University of Toronto, Toronto, ON, Canada

W. D. Foulkes � P. N. Tonin

Program in Cancer Genetics, Departments of Oncology and

Human Genetics, McGill University, Montreal, QC, Canada

W. D. Foulkes

Department of Medical Genetics, Sir Mortimer B. Davis-Jewish

General Hospital, Montreal, QC, Canada

W. D. Foulkes � P. N. Tonin

Departments of Medicine and Human Genetics, McGill

University, Montreal, QC, Canada

D. Provencher

Departement d’obstetrique gynecologie, Division de gynecologie

oncologique, Universite de Montreal, Montreal, QC, Canada

P. N. Tonin (&)

Medical Genetics, Montreal General Hospital, Room L10-120,

Montreal, QC, Canada H3G 1A4

e-mail: [email protected]

123

Breast Cancer Res Treat

DOI 10.1007/s10549-007-9608-6

selected for family history of cancer. In all, germline TP53

mutations were identified in two of 52 (3.8%) cancer

families, suggesting that TP53 is not a major contributor to

BRCA1 and BRCA2 mutation-negative breast and/or

ovarian cancer families of French Canadian descent.

Keywords TP53 � BRCA1 � BRCA2 � Breast cancer �French Canadian � Founder effects

Introduction

Approximately 40% of French Canadian families from the

Province of Quebec containing at least three cases of fe-

male breast cancer (diagnosed <65 years of age), ovarian

cancer, or male breast cancer have been shown to harbor

germline mutations in BRCA1 and BRCA2 [1–3]. Specific

BRCA1 and BRCA2 mutations have been found to recur in

French Canadian cancer families [2–5], and this has been

attributed to common founders in the Quebec population

[1, 6–10]. Mutation-negative families could be due to

unidentified BRCA1/BRCA2 mutations, chance clustering

of cancer cases or other genes conferring increased risk to

breast cancer [11].

Breast cancer has also been reported in Li–Fraumeni

Syndrome (LFS) families. LFS is a rare autosomal disorder

characterized by familial clustering of tumors, with a pre-

dominance of sarcomas, breast cancers, brain tumors and

adrenocortical carcinomas, diagnosed before the age of

45 years, where about 70% of families have been shown to

harbor germline mutations in TP53 [12–17]. Other cancers,

such as leukemia, lung cancer, skin melanoma, gastric

cancer, pancreatic cancer, and prostate cancer are also in

excess in some families; and rare cancers such as germ cell

tumors, choroid plexus papilloma, and Wilms tumor have

been reported [14, 18–20]. However, not all of the families

with germline TP53 mutations fulfill the definition of

classical LFS and Li–Fraumeni syndrome-like (LFL) has

been used to describe families which share some features

consistent with those described in LFS families [13, 16,

21]. LFS families are characterized by a proband with

sarcoma diagnosed <45 years of age; a first-degree relative

with any cancer diagnosed <45 years of age; and a first- or

second-degree relative in the same lineage diagnosed with

a sarcoma at any age or any cancer diagnosis occurring

<45 years of age [12]. Birch et al. [13] have defined LFL

families as a proband with any childhood cancer or sar-

coma, brain tumor or adrenal cortical tumor diagnosed

before 45 years of age and a first- or second-degree relative

with a typical LFS cancer (sarcoma, breast cancer, brain

tumor, adrenal cortical tumor or leukemia) at any age, and

a first- or second-degree relative with any cancer under the

age of 60 years [13]. Another definition of LFL is the

occurrence of two first- or second-degree relatives with

LFS-related malignancies at any age [22].

A feature in common with LFS and LFL families is the

presence of very young age of onset breast cancers, which

is a feature that overlaps hereditary breast cancer (HBC)

and breast-ovarian cancer (HBOC) families found to harbor

germline BRCA1 and BRCA2 mutations [23, 24]. The

frequency of TP53 germline mutations has been investi-

gated in multiple studies [25–32], suggesting that they

account for <1% of breast cancer families. However, the

contribution of TP53 to breast cancer in the French

Canadian population is not known. In the present study we

describe the frequency of a TP53 mutation, 13398 G>A

(Arg213Gln), identified in a family of French Canadian

descent originally selected for inclusion in a HBC study.

Given the strong founder effect in this population, we

determined the frequency of this mutation in a series of 52

independently ascertained HBC or HBOC families previ-

ously tested and found negative for BRCA1 and BRCA2

mutations. We also performed mutation analysis of all

domain regions and splice sites of TP53 in this series of

families and report on the deleterious mutation and variants

identified. The frequency of the deleterious mutations

identified was investigated in a series of breast cancer cases

diagnosed <50 years of age from French Canadian women

not selected for family history of breast cancer.

Materials and methods

Cancer families

TP53 mutation analysis was performed on index cases from

52 HBC and HBOC cancer families (Table 1). Each family

had at least three confirmed cases of female breast cancer

(diagnosed £ 65 years of age), epithelial ovarian cancer, or

male breast cancer. The affected individuals in each family

were first-, second- or third-degree relatives (occurring

within the same lineage) to the index case that was selected

for TP53 mutation analysis. Index cases reported grandpa-

rental French Canadian ancestry from Quebec, Canada.

Index cases from 42 of the 52 families were previously found

negative for BRCA1 and BRCA2 mutations by commercial

sequencing service (Myriad Genetics�, http://www.

myriad.com/index.php). Index cases from the remaining ten

families in the series analyzed for TP53 mutations were

found negative for sequence analysis of eight of the most

common BRCA1 (2953delGTAinsC, 3875delGTCT,

4446C>T, and 5221delTG) and BRCA2 (3398delAAAAG,

6085G>T, 7235G>A, and 8765delAG) mutations identified

in French Canadian cancer families that together account for

about 84% of the mutation-positive cases with a family

history exhibiting the inclusion criteria described above


123

http://www.myriad.com/index.php

http://www.myriad.com/index.php

[1–3]. The families were ascertained through the Service de

Medecine Genique, Centre Hospitalier de l’Universite de

Montreal (CHUM) and the Hereditary Cancer Clinics of

McGill University in Montreal.

Breast cancer cases

The 381 female breast cancer cases tested for the pres-

ence of deleterious TP53 mutations were initially ascer-

tained as part of a hospital-based study of breast cancer

susceptibility among French Canadian women. The par-

ticipants were diagnosed with invasive breast cancer at or

under the age of 50 years at a single hospital at the Breast

Clinic of the CHUM—Hotel-Dieu Hospital, Montreal.

The participants were not selected for family history of

cancer, and were BRCA1 and BRCA2 mutation-negative.

The participants identified themselves to be of French

Canadian ancestry. The average age of diagnosis was

44 years (range 26–50 years). Within this series there

were 78 breast cancer cases diagnosed before age

40 years. None of the cases are known to be related to

each other.

TP53 mutation analyses

DNA was extracted from peripheral blood lymphocytes.

Polymerase chain reaction (PCR) was used to amplify the

domain regions and flanking splice sites of exons 2–11 of

TP53, using primer sets and conditions described in

Table 2. PCR was performed in a 25 ll volume con-

taining 200 ng of genomic DNA; 1· PCR buffer (Invi-

trogen, Burlington, Canada); 5 nmol each dCTP, dGTP,

dTTP, and dATP; 1.5 mM MgCl2; 15 pmol of each pri-

mer; and 1 U of Taq polymerase (Invitrogen). The PCR

conditions were 3 min at 95�C, 35 cycles of 94�C for

30 s, appropriate annealing temperature (Table 2) for 30 s

and 72�C for 30 s. The PCR products were sequenced

using the 3730XL DNA Analyzer system platform from

Applied Biosystems at the McGill University and Gen-

ome Quebec Innovation Center, Montreal, Canada (ge-

nomequebec.mcgill.ca). Sequence chromatograms were

compared with the TP53 reference sequence X54156 and

the TP53 genomic sequence represented in the UCSC

Genome Bioinformatics Human Genome Browser Gate-

way March 2006 hg 18 assembly (genome.ucsc.edu/cgi-

bin/hgGateway).

The 381 female breast cancer cases not selected for

family history of cancer were investigated for the presence

of the TP53 13398G>A and 14538G>A mutations identi-

fied in the French Canadian breast cancer families. Geno-

mic DNA was extracted from peripheral lymphocytes.

Mutation analysis was performed using the tetra-primer

Amplification Refractory Mutation System (ARMS) PCR

method as described previously [33].

Results

Frequency of TP53:13398 G>A mutation

The TP53:13398 G>A mutation (Table 3) was initially

identified in a women diagnosed with breast cancer at age

41 from Family 1444. This individual was tested for

mutations in TP53 because of close relatives with very

young age of diagnoses of brain tumors and an adrenal

cortical carcinoma, which are features consistent with LFS

(Fig. 1). The index case was previously tested and found

negative for BRCA1 and BRCA2 mutations as part of a

study of French Canadian HBC families [3]. Notable are

the six breast cancer cases in first- and second-degree rel-

atives of the index case, which include four cases diag-

nosed at <55 years of age. TP53 mutation analysis of

Table 1 Features of index cases

Syndrome Number of

families

Primary cancer of index case

Primary

site

Mean

age

Median

age

Age

range

Number with multiple

cancers

Cases with mulitiple Br

and/or Ov

Cases with other

cancers

HBC 34 Br 46.2 45.5 30–64 10 BiBr38 Br36Leu41

BiBr39–42 Br37Leu52

BiBr47 Br54Lu59Lar60

BiBr53–54 Br47Mel40

Bi59–66 Br60Sk62–64

HBOC 14 Br 48.6 38 38–64 3 Br38Per75

BiBr40–45

BiBr42–43

HBOC 4 Ov 40.3 37.5 33–42 1 Ov42Br45


123

family members indicated inheritance from the maternal

branch of the family of the index case.

Given the founder effects in French Canadian popula-

tion, the frequency of 13398 G>A mutation was deter-

mined by screening HBC and HBOC families previously

tested negative for BRCA1 and BRCA2 mutations (Ta-

ble 1). The analysis of index cases of 52 families identified

only one carrier of this mutation. The mutation carrier,

identified in Family 1039 (Fig. 1), was diagnosed with

bilateral breast cancer at age 38. DNA was not available

from other cases in this family and thus the carrier status of

other individuals in this family is not known. Notable is

that the features of this family are not consistent with those

found in either LFS or LFL families. The 13398 G>A

mutation was not detected in 381 French Canadian women

diagnosed with breast cancer before the age of 50 years not

selected for family history of cancer.

Mutation analysis of TP53

The identification of germline TP53 mutation in HBC

family prompted a mutation analysis of the domain regions

and splice sites of this gene in the remaining HBC and

HBOC families tested negative for BRCA1 and BRCA2

mutations (Table 1). This mutation analysis identified 30

sequence variants (Table 3). The majority of variants occur

within intronic sequences, are predicted to have no effect

on the functionality of TP53, and have been classified as

polymorphisms in the IARC TP53 Mutation Database

(www-p53.iarc.fr) and/or the Single Nucleotide Polymor-

phism (SNP) Database (www.ncbi.nlm.nih.gov/projects/

SNP/). Although the genotype and allele frequencies of

these variants in the French Canadian population are not

known, those exhibited by the index cases are comparable

to frequencies reported in the SNP Database and/or the

Cancer Genome Anatomy Project SNP500Cancer Database

(snp500cancer.nci.nih.gov) (Table 3).

Of the four variants predicted to affect the encoded

product due to a missense mutation, only the 14538 G>A

sequence variation resulting in the Arg290His amino acid

substitution has previously been classified as a mutation in

the IARC TP53 Database. This mutation was identified in

an index case diagnosed with breast cancer at age 44 years

in Family 875 (Fig. 1). Although there were multiple cases

of cancer reported in this family, the features are not

consistent with those seen in LFS or LFL families. Notable

is that this mutation was not detected in a maternal aunt

diagnosed with breast cancer at age 61 and colon cancer at

age 81. DNA was not available from other family members

and thus the carrier status of the other cancer cases in this

family is not known. The 14538 G>A mutation was also

not detected in 381 French Canadian women diagnosed

with breast before the age of 50 years not selected for

family history of cancer.

Discussion

Germline mutations in TP53 were identified in two of 52

(3.8%) BRCA1 and BRCA2 mutation-negative families of

French Canadian descent. The frequency of mutation-po-

sitive families identified in our study is consistent with

independent reports suggesting that germline TP53 muta-

tions are rare in multiple case breast cancer families, pos-

sibly accounting for <1% of such families [25–32, 34–37].

The higher frequency of mutation-positive families in our

study is likely due to the selection criteria, which excluded

BRCA1 and BRCA2 mutation-positive families. The

Table 2 Primer sequences for

amplification and/or sequencing

a Genomic region amplified

and/or sequenced based on the

Human Genome Browser

Gateway March 2006 (hg18)

assembly described in the

UCSC Genome Bioinformatics

database (genome.ucsc.edu/cgi-

bin/hgGateway)b Primer sequences used for

DNA sequencing only

TP53 exon Genomic regiona Primer sequences Annealing

temperature (�C)

2, 3, and 4 7519877–7520827 F: 5¢-tggaagagagaatgtgaagc-3¢ 60

R: 5¢-caggagtcagagatcacacat-3¢5 and 6 7518762–7519376 F: 5¢-gcttacgcatgtttgtttct-3¢ 55

R: 5¢-acccatttactttgcacatc-3¢7 7518106–7518440 F: 5¢-gacagagcgagattccatc-3¢ 63.5

R: 5¢-tgagaggtggatgggtagta-3¢7b 7518127–7518394 F: 5¢-tgcttgccacaggtctccc-3¢ NA

R: 5¢-tatggaagaaatcggtaagag-3¢8 and 9 7517517–7517967 F: 5¢-gagtagatggagcctggttt-3¢ 55

R: 5¢-cggcattttgagtgttagac-3¢10 7514528–7514895 F: 5¢-taggtacttgaagtgcagtttct-3¢ 55

R: 5¢-ctgggacccaatgagatg-3¢11 7513367–7513852 F: 5¢-ttgatttgaattcccgttg-3¢ 55

R: 5¢-aacccttaactgcaagaaca-3¢


123

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9.0

1.0

96

–


123

identification of two 13398 G>A mutation-positive fami-

lies is not attributable to recurrent mutations in this founder

population as subsequent pedigree expansion revealed that

the index cases of the two independently ascertained

families are cousins (Fig. 1).

The 13398 G>A (Arg213Gln) and 14538 G>A (Ar-

g290His) mutations are both missense mutations and would

affect the amino acid composition of the non-DNA-binding

loops in the beta-sheet scaffold of the p53 DNA binding

domain (www-p53.iarc.fr). Both mutations have been re-

ported in the IARC TP53 Mutation Database occurring as

both somatic and germline events. However, based on

various prediction tools for amino acid substitutions, the

IARC TP53 Mutation Database has classified the 13398

G>A nucleotide substitution as a deleterious mutation and

the 14538 G>A nucleotide substitution as a neutral change.

Notable is that the 13398 G>A mutation appears to seg-

regate with affected cases in Family 1444 which contained

features consistent with LFS, such as the pediatric cases of

adrenocortical carcinoma and brain tumors occurring in

first- and second-degree relatives of the index case (Fig. 1).

The mutated allele was maternally transmitted, and it was

through pedigree expansion of this maternal branch of the

family that the index case was linked to another who car-

ried of the same mutation, but was ascertained indepen-

dently from Family 1039 (Fig. 1). The number and age

range of the breast cancer cases with diagnosis before age

55 years in Family 1444 overlap familial cases of breast

cancer found to harbor either BRCA1 and BRCA2 germ-

line mutations ascertained from the French Canadian

population of Quebec [1–3]. However, the mother of the

index case was diagnosed with basal cell carcinoma at age

66 years, followed by diagnoses of breast and pancreatic

cancers at age 69. The late age of diagnosis of cancers in

this mutation carrier is interesting in light of younger ages

of diagnoses of cancer and severity of disease in her chil-

dren and grandchildren. Her sister was also diagnosed with

similar late age of onset cancers, a breast cancer at age 74

and a pancreatic carcinoma at age 75. The two other sisters

who were both diagnosed with breast cancer at age

53 years. Recently, it was suggested that the 13398 G>A

mutation confers susceptibility to late age of onset cancers

by the genetic analysis of a large kindred with multiple and

varied adult onset cancers [38]. However, in Family 1444,

the family history of cancer, carrier status of tested cases,

and probable cases of pediatric cancers in two other

branches of the family, would imply that this mutant alleleTa

ble

3co

nti

nu

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atio

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ele

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a

AA

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Po

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xl1

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ared

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(Cau

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54

15

6

Fig. 1 Pedigrees of TP53 mutation-positive families. Cancer type or

site [adrenocorticol carcinoma (ACC), bladder (Bla), brain (Bt), breast

(Br), cervical (Cx), colon (Co), kidney (Ki), liver (Li), lung (Lu)

pancreatic (Pan), skin (Sk)] and age of diagnosis are shown or age of

death (d) along with TP53 mutation carrier status. The index case

initially tested for TP53 mutation is shown with an arrow


123

Family 1444

IV

III

II

I

31Bt 6d. 9

3233ACC 9

2627Br 41

+

29Bt 12

+

30

24

-

25Br 43d. 45

21Bt 1d. 1

9d. 45

10

19ACC ?

20

5

16BiBr 38

17

3Br 53Li 76d. 76

28

13

-

14Sk 66Br 69

Pan 69

+

8Br 74

Pan 75d. 76

11Br 53d. 56

15

1d. 56

2d. 66

4

Family 875

III

II

I

21Bla 33d. 38

9 10

1920Br 44

+

673Br 61Co 81d. 83

-

4d. 28

5Br 62

8Bla 65Psu 73d. 74

11d. 11

12Pan 74d. 74

13Lu 65

Pan 68d. 69

14Kd 66d. 66

15 16

1d. 85

2Br 68d. 75

2

18 2322

6124

Family 1039

III

II

I

8Cx 49

9 10BiBr 38

11 12

3LuLeu

4Br 53

5Br 53

6 7

1 2

6 3

3

+


123

is highly penetrant warranting clinical surveillance for

family members.

The significance of 14538 G>A mutation is less certain.

Family 875 does not exhibit features consistent with LFS

or LFL (Fig. 1). Interestingly, Canada was reported as the

country of origin for the two 14538 G>A mutation-positive

families noted in the IARC TP53 Mutation Database.

Based on the reference citation in the database, both

mutation-positive families were initially described by

Quesnel et al. [39]. A review of the reported pedigrees

[39], shows that the 14538 G>A mutation carrier probands

are associated with an LFS and LFL families. The proband

in the LFS family was diagnosed with a soft tissue sarcoma

at age two and then a brain tumor at age 10. However, one

of these probands also carried two other TP53 variants,

which affected codons 156 (CGC>CAC, Arg156His) and

267 (CGG>CAG, Arg267Gln). Genotyping analysis indi-

cated that the index case inherited the compound mutation

from the mother who was affected with bilateral breast

cancer (at ages 35 and 43 years), whereas the father who

carried and transmitted the 14538 G>A allele was unaf-

fected with cancer. Although both codon 156 and 267

mutations would affect the p53 DNA binding domain, only

the codon 267 mutation is classified as deleterious in the

IARC TP53 Mutation Database. However, various in vitro

assays assessing the functionality of the TP53 variants

suggest that all variants exhibited partial or complete loss

of wild-type p53 function, where the 14538 G>A alteration

exhibited the least detrimental effects overall in compari-

son to the other mutations alone or combined [39]. Neither

parent of the 14538 G>A mutation carrier proband in the

LFL family reported a personal history of cancer, although

a paternal grandfather was diagnosed with a brain tumor at

age 40 years and a paternal cousin was diagnosed with a

rhabdomyosarcoma at 4 years of age (with both parents

also unaffected with cancer) [39]. Although other members

of this family were not tested for carrier status, the family

history would imply incomplete penetrance. This is also

suggested by the family history and carrier status in the

14538 G>A mutation-positive family identified in our

study. The 14538 G>A mutation-positive index case in

Family 875 did not exhibit features consistent with LFS or

LFL. Moreover, the one other cancer case available for

testing (a maternal aunt with breast cancer at 61 and colon

cancer at 81) tested negative for this mutation (Fig. 1).

However, it is not known if this mutant allele was trans-

mitted from the maternal branch of the family, which had

cancer cases, or the paternal branch where the family his-

tory of cancer is unknown, as DNA is not available from

the parents of the index test in this family.

The low frequency of germline TP53 mutations in HBC

and HBOC families of French Canadian descent would

suggest that routine screening for TP53 mutations in

BRCA1 and BRCA2 mutation-negative families is not

warranted. The observation that the TP53 mutations were

not found in a series of 381 invasive breast cancer cases

diagnosed before age 50 years would suggest that these

variants are not present in excess in this founder population

and thus screening for these mutations is also not war-

ranted. However, during the course of this study indepen-

dent reports appeared showing that germline mutations in

novel candidate breast cancer susceptibility genes ATR

[40] and PALB2 [41] were not identified in French Cana-

dian HBC and HBOC families previously tested negative

for BRCA1 and BRCA2 mutations. Although germline

PALB2 mutations have been reported in breast cancer

families [42], this gene appears not to be a major con-

tributor to HBC in the French Canadian cancer families

[41].

The identification of germline TP53 mutations in HBC

families is disconcerting. Careful monitoring of such

families could over time reveal features consistent with

those found in LFS or LFL families, as demonstrated with

families 1444 and 1039. The clinical significance of the

13398 G>A mutation is evident in the family history of

cancer in Family 1444 and monitoring of family members

is warranted. Less certain is the significance of 14538 G>A

mutation classified as neutral by the IARC TP53 Database

and identified in a site-specific breast cancer family, but

found independently to occur in a LFS family. It is not

clear if this mutation segregates with cancer in both inde-

pendently ascertained families. Although both indepen-

dently ascertained families with this mutation were

identified in Canada, the familial relationship is unknown.

Given the founder effects in the French Canadian popula-

tion it would be interesting to determine if the carriers of

the 14538 G>A mutation are identical-by-descent. Further

characterization the TP53 14538 G>A mutation is war-

ranted in light of the association of affected carriers with

cancer families featuring different cancer types.

Acknowledgments With thank Marise Roy, Stephanie Lepage,

Manon Deladurantaye, and Myriam Costa for technical support.

C.M.M. is a recipient of a chercheur-boursier Fonds de la recherche

en sante du Quebec (F.R.S.Q.). This work was supported by grants

from the Cancer Research Society Inc., to P.N.T., and the Banque de

tissus et de donnees of the Reseau de recherche sur le cancer of the

F.R.S.Q. to W.D.F., P.G, C.M.M., A.M. M.-M., D.M.P., and P.N.T.

References

1. Tonin PN, Mes-Masson AM, Futreal PA, Morgan K, Mahon M,

Foulkes WD, Cole DE, Provencher D, Ghadirian P, Narod SA

(1998) Founder BRCA1 and BRCA2 mutations in French

Canadian breast and ovarian cancer families. Am J Hum Genet

63(5):1341–1351

2. Oros KK, Ghadirian P, Greenwood CM, Perret C, Shen Z,

Paredes Y, Arcand SL, Mes-Masson AM, Narod SA, Foulkes


123

WD et al (2004) Significant proportion of breast and/or ovarian

cancer families of French Canadian descent harbor 1 of 5 BRCA1

and BRCA2 mutations. Int J Cancer 112(3):411–419

3. Oros KK, Ghadirian P, Maugard CM, Perret C, Paredes Y, Mes-

Masson AM, Foulkes WD, Provencher D, Tonin PN (2006)

Application of BRCA1 and BRCA2 mutation carrier prediction

models in breast and/or ovarian cancer families of French

Canadian descent. Clin Genet 70(4):320–329

4. Antoniou AC, Durocher F, Smith P, Simard J, Easton DF (2006)

BRCA1 and BRCA2 mutation predictions using the BOADICEA

and BRCAPRO models and penetrance estimation in high-risk

French–Canadian families. Breast Cancer Res 8(1):R3

5. Simard J, Dumont M, Moisan AM, Gaborieau V, Vezina H,

Durocher F, Chiquette J, Plante M, Avard D, Bessette P et al

(2007) Evaluation of BRCA1 and BRCA2 mutation prevalence,

risk prediction models and a multi-step testing approach in

French–Canadian high-risk breast and ovarian cancer families. J

Med Genet 44(2):107–121

6. Manning AP, Abelovich D, Ghadirian P, Lambert JA, Frappier D,

Provencher D, Robidoux A, Peretz T, Narod SA, Mes-Masson

AM et al (2001) Haplotype analysis of BRCA2 8765delAG

mutation carriers in French Canadian and Yemenite Jewish

hereditary breast cancer families. Hum Hered 52(2):116–120

7. Scriver CR (2001) Human genetics: lessons from Quebec popu-

lations. Annu Rev Genomics Hum Genet 2:69–101

8. Laberge AM, Michaud J, Richter A, Lemyre E, Lambert M, Brais

B, Mitchell GA (2005) Population history and its impact on

medical genetics in Quebec. Clin Genet 68(4):287–301

9. Vezina H, Durocher F, Dumont M, Houde L, Szabo C, Tranchant

M, Chiquette J, Plante M, Laframboise R, Lepine J et al (2005)

Molecular and genealogical characterization of the R1443X

BRCA1 mutation in high-risk French–Canadian breast/ovarian

cancer families. Hum Genet 117(2–3):119–132

10. Oros KK, Leblanc G, Arcand SL, Shen Z, Perret C, Mes-Masson

AM, Foulkes WD, Ghadiran P, Provencher D, Tonin PN (2006)

Haplotype analysis suggest common founders in carriers of the

recurrent BRCA2 mutation, 3398delAAAAG, in French Cana-

dian hereditary breast and/ovarian cancer families. BMC Med

Genet 7(1):23

11. Antoniou AC, Easton DF (2006) Models of genetic susceptibility

to breast cancer. Oncogene 25(43):5898–5905

12. Li FP, Fraumeni JF Jr, Mulvihill JJ, Blattner WA, Dreyfus MG,

Tucker MA, Miller RW (1988) A cancer family syndrome in

twenty-four kindreds. Cancer Res 48(18):5358–5362

13. Birch JM, Hartley AL, Tricker KJ, Prosser J, Condie A, Kelsey

AM, Harris M, Jones PH, Binchy A, Crowther D et al (1994)

Prevalence and diversity of constitutional mutations in the p53

gene among 21 Li-Fraumeni families. Cancer Res 54(5):1298–

1304

14. Frebourg T, Barbier N, Yan YX, Garber JE, Dreyfus M, Frau-

meni J Jr, Li FP, Friend SH (1995) Germ-line p53 mutations in 15

families with Li-Fraumeni syndrome. Am J Hum Genet

56(3):608–615

15. Varley JM, McGown G, Thorncroft M, Santibanez-Koref MF,

Kelsey AM, Tricker KJ, Evans DG, Birch JM (1997) Germ-line

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

39 families. Cancer Res 57(15):3245–3252

16. Evans DG, Birch JM, Thorneycroft M, McGown G, Lalloo F,

Varley JM (2002) Low rate of TP53 germline mutations in breast

cancer/sarcoma families not fulfilling classical criteria for Li-

Fraumeni syndrome. J Med Genet 39(12):941–944

17. Olivier M, Goldgar DE, Sodha N, Ohgaki H, Kleihues P, Hainaut

P, Eeles RA (2003) Li-Fraumeni and related syndromes: corre-

lation between tumor type, family structure, and TP53 genotype.

Cancer Res 63(20):6643–6650

18. Strong LC, Stine M, Norsted TL (1987) Cancer in survivors of

childhood soft tissue sarcoma and their relatives. J Natl Cancer

Inst 79(6):1213–1220

19. Garber JE, Burke EM, Lavally BL, Billett AL, Sallan SE, Scott

RM, Kupsky W, Li FP (1990) Choroid plexus tumors in the

breast cancer-sarcoma syndrome. Cancer 66(12):2658–2660

20. Hartley AL, Birch JM, Tricker K, Wallace SA, Kelsey AM,

Harris M, Jones PH (1993) Wilms’ tumor in the Li-Fraumeni

cancer family syndrome. Cancer Genet Cytogenet 67(2):133–135

21. Eeles RA, Bartkova J, Lane DP, Bartek J (1993) The role of TP53

in breast cancer development. Cancer Surv 18:57–75

22. Eeles RA (1995) Germline mutations in the TP53 gene. Cancer

Surv 25:101–124

23. Malkin D, Li FP, Strong LC, Fraumeni JF Jr, Nelson CE, Kim

DH, Kassel J, Gryka MA, Bischoff FZ, Tainsky MA et al (1990)

Germ line p53 mutations in a familial syndrome of breast cancer,

sarcomas, and other neoplasms. Science 250(4985):1233–1238

24. Garber JE, Goldstein AM, Kantor AF, Dreyfus MG, Fraumeni JF

Jr, Li FP (1991) Follow-up study of twenty-four families with Li-

Fraumeni syndrome. Cancer Res 51(22):6094–6097

25. Prosser J, Elder PA, Condie A, MacFadyen I, Steel CM, Evans HJ

(1991) Mutations in p53 do not account for heritable breast

cancer: a study in five affected families. Br J Cancer 63(2):181–

184

26. Borresen AL, Andersen TI, Garber J, Barbier-Piraux N, Thorla-

cius S, Eyfjord J, Ottestad L, Smith-Sorensen B, Hovig E, Malkin

D et al (1992) Screening for germ line TP53 mutations in breast

cancer patients. Cancer Res 52(11):3234–3236

27. Warren W, Eeles RA, Ponder BA, Easton DF, Averill D, Ponder

MA, Anderson K, Evans AM, DeMars R, Love R et al (1992) No

evidence for germline mutations in exons 5–9 of the p53 gene in

25 breast cancer families. Oncogene 7(5):1043–1046

28. Jolly KW, Malkin D, Douglass EC, Brown TF, Sinclair AE, Look

AT (1994) Splice-site mutation of the p53 gene in a family with

hereditary breast-ovarian cancer. Oncogene 9(1):97–102

29. Sun XF, Johannsson O, Hakansson S, Sellberg G, Nordenskjold

B, Olsson H, Borg A (1996) A novel p53 germline alteration

identified in a late onset breast cancer kindred. Oncogene

13(2):407–411

30. Rapakko K, Allinen M, Syrjakoski K, Vahteristo P, Huusko P,

Vahakangas K, Eerola H, Kainu T, Kallioniemi OP, Nevanlinna

H et al (2001) Germline TP53 alterations in Finnish breast cancer

families are rare and occur at conserved mutation-prone sites. Br

J Cancer 84(1):116–119

31. Martin AM, Kanetsky PA, Amirimani B, Colligon TA, Athana-

siadis G, Shih HA, Gerrero MR, Calzone K, Rebbeck TR, Weber

BL (2003) Germline TP53 mutations in breast cancer families

with multiple primary cancers: is TP53 a modifier of BRCA1? J

Med Genet 40(4):e34

32. Walsh T, Casadei S, Coats KH, Swisher E, Stray SM, Higgins J,

Roach KC, Mandell J, Lee MK, Ciernikova S et al (2006)

Spectrum of mutations in BRCA1, BRCA2, CHEK2, and TP53 in

families at high risk of breast cancer. Jama 295(12):1379–1388

33. Ye S, Dhillon S, Ke X, Collins AR, Day IN (2001) An efficient

procedure for genotyping single nucleotide polymorphisms. Nu-

cleic Acids Res 29(17):E88–8

34. Lehman TA, Haffty BG, Carbone CJ, Bishop LR, Gumbs AA,

Krishnan S, Shields PG, Modali R, Turner BC (2000) Elevated

frequency and functional activity of a specific germ-line p53

intron mutation in familial breast cancer. Cancer Res 60(4):1062–

1069

35. Balz V, Prisack HB, Bier H, Bojar H (2002) Analysis of BRCA1,

TP53, and TSG101 germline mutations in German breast and/

or ovarian cancer families. Cancer Genet Cytogenet 138(2):120–

127


123

36. Bendig I, Mohr N, Kramer F, Weber BH (2004) Identification of

novel TP53 mutations in familial and sporadic cancer cases of

German and Swiss origin. Cancer Genet Cytogenet 154(1):22–26

37. Lalloo F, Varley J, Moran A, Ellis D, O’Dair L, Pharoah P,

Antoniou A, Hartley R, Shenton A, Seal S et al (2006) BRCA1,

BRCA2 and TP53 mutations in very early-onset breast cancer

with associated risks to relatives. Eur J Cancer 42(8):1143–1150

38. Ruijs MW, Verhoef S, Wigbout G, Pruntel R, Floore AN, de Jong

D, van TVLJ Menko FH (2006) Late-onset common cancers in a

kindred with an Arg213Gln TP53 germline mutation. Fam Can-

cer 5(2):169–174

39. Quesnel S, Verselis S, Portwine C, Garber J, White M, Feunteun

J, Malkin D, Li FP (1999) p53 compound heterozygosity in a

severely affected child with Li-Fraumeni syndrome. Oncogene

18(27):3970–3978

40. Durocher F, Labrie Y, Soucy P, Sinilnikova O, Labuda D, Bes-

sette P, Chiquette J, Laframboise R, Lepine J, Lesperance B et al

(2006) Mutation analysis and characterization of ATR sequence

variants in breast cancer cases from high-risk French Canadian

breast/ovarian cancer families. BMC Cancer 6:230

41. Tischkowitz M, Xia B, Sabbghian N, Reis-Filho JS, Hamel N, Li

G, van Beers EK, li L, Khalil T, Quenneville LA, Omeroglu A,

Poll A, Lepage P, Wong N, Nederlof PM, Ashworth A, Tonin

PN, Narod SA, Livingston DM, Foulkes WD (2007) Analysis of

PALB2/FANCN-associated breast cancer families. PNAS

104(16):6788–6793

42. Rahman N, Seal S, Thompson D, Kelly P, Renwick A, Elliott A,

Reid S, Spanova K, Barfoot R, Chagtai T et al (2007) PALB2,

which encodes a BRCA2-interacting protein, is a breast cancer

susceptibility gene. Nat Genet 39(2):165–167


123

Germline TP53 mutations in BRCA1 and BRCA2 mutation-negative French Canadian breast cancer families

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