Variation of p53 Mutational Spectra between Carcinoma of ... · Vol. 1, 763-768, Jo/v 1995 Clinical...
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Vol. 1, 763-768, Jo/v 1995 Clinical Cancer Research 763
3 The abbreviations used are: SCCHN, squamous cell carcinoma of the
head and neck; URT, upper respiratory tract; LRT, lower respiratory tract.
Variation of p53 Mutational Spectra between Carcinoma of the
Upper and Lower Respiratory Tract’
John C. Law,2 Theresa L. Whiteside,
Susanne M. Gollin, Joel Weissfeld,
Lobna El-Ashmawy, S. Srivastava,
Rodney J. Landreneau, Jonas T. Johnson,
and Robert E. Ferrell
Departments of Human Genetics [J. C. L., S. M. G., R. E. F.J and
Epidemiology [J. W.J, Graduate School of Public Health, University
of Pittsburgh and Pittsburgh Cancer Institute, Pittsburgh 15261;
Departments of Pathology [T. L. W., L. E-A.], Surgery [R. J. L.], and
Otolaryngology [J. T. J.], University of Pittsburgh Medical Center
and Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania 15261 ; and
National Cancer Institute, Division of Cancer Prevention and Control,
Early Detection Branch, NIH, Bethesda, Maryland 20892 [S. S.]
ABSTRACT
Mutations of the p5.3 tumor suppressor gene are the most
common genetic alterations associated with human cancer.
Tumor-associated p5.3 mutations often show characteristic tis-
sue-specific profiles which may infer environmentally induced
mutational mechanisms. The p5.3 mutational frequency and
spectrum were determined for 95 carcinomas of the upper and
lower respiratory tract (32 lung and 63 upper respiratory
tract). Mutations were identified at a frequency of 30% in
upper respiratory tract (URT) tumors and 31 % in lung tu-mors. All 29 identified mutations were single-base substitu-
tions. Comparison of the frequency of specific base substitu-
tions between lung and URT showed a striking difference.
Transitions occurred at a frequency of 68% in URT, but only
30% in lung. Mutations involving G:C-�A:T transitions,which are commonly reported in gastric and esophageal hi-
mors, were the most frequently identified alteration in URT
(11119). Mutations involving G:C-*T:A transversions, which
were relatively common in lung tumors (3/10) and are repre-
sentative of tobacco smoke-induced mutations were rare in
URT tumors (1/19). Interestingly, G:C-�A:T mutations at
CpG sites, which are characteristic of endogenous processes,
were observed frequently in URT tumors (9/19) but only rarely
in lung tumors (1/10), suggesting that both endogenous and
exogenous factors are responsible for the observed differences
in mutational spectra between the upper and lower respiratory
systems.
Received 12/19/94; accepted 3/23/95.
1 This work was supported by NCI(MAO)CN-15393-02 and in part by
NCI-CN-24428-33, ACS Grant EDT-44, the Mary Hillman Jennings
Foundation, and the John R. McCune Charitable Trust Foundation.
2 To whom requests for reprints should be addressed, at Department ofHuman Genetics, Graduate School of Public Health, 130 DeSoto Street,
University of Pittsburgh, Pittsburgh, PA 15261.
INTRODUCTION
Two common cancers with poor prognosis, high mortality,
and clearly defined and overlapping environmental exposure
risk factors are SCCHN3 and lung cancer. SCCHN is the sixth
most frequent cancer worldwide (1). In the United States, ap-
proximately 11,000 individuals die annually from SCCHN (2).
There has been little improvement in the prognosis for SCCHN
oven the past 30 years, and it remains relatively poor, with an
overall 5-year survival rate of 54% (2, 3). Lung cancer is the
leading form of cancer diagnosed in the United States with an
overall incidence rate of 55.2/100,000 population (4). Cigarette
smoking is widely accepted as the major risk factor for the
development of lung cancer, with 80% of lung cancer incidence
attributed to exposure to tobacco smoke (5, 6). SCCHN shares
tobacco smoking as a common major risk factor with lung
cancer, although exposure to smokeless tobacco and alcohol are
additional significant risk factors for SCCHN (7, 8).
Mutations of the p53 tumor suppressor gene are the most
common genetic alteration associated with human cancer. Cur-
rent evidence suggests that the wild-type p53 protein is essential
for normal cell growth regulation and that its alteration or
inactivation is associated with the development of cancer (9). A
major function of the p53 gene is believed to be as a cell cycle
check point gene (10). Thep53 gene is induced by DNA damage
with a resultant transient cell cycle arrest at the G1-S interface
(1 1-13). Cells lacking wild-type p53 do not display this DNA
damage-induced cell cycle arrest (14, 15). The p53 gene has
recently been shown to act as a transactivator of a cell cycle-
associated protein that directly interacts with cyclin-dependent
kinases involved in G1 arrest (16, 17).
Many different point, deletion, and insertion mutations
have been described which can inactivate p53-mediated tumor
suppression (18, 19). The analysis of tumor DNA has revealed
that p53 mutations are usually missense mutations which lead to
amino acid substitutions in the protein, and are primarily found
in one of four evolutionanily conserved regions of nucleotide
sequence located in exons 5-8 (20, 21). Mutations of the p53
tumor suppressor gene are common in both lung cancer (22-26)
and in SCCHN (27-30).
The multistep process of carcinogenesis requires an accu-
mulation of multiple genetic alterations in order for normal cells
to progress to cancer. The role of mutations in the p53 tumor
suppressor gene in human cancer has been well established.
DNA damage may be induced by a number of factors including
endogenous metabolites and exogenous chemical and physical
carcinogenic compounds. It is known that various carcinogens
can induce specific DNA base changes (31, 32). Tissue-specific
mutational spectra may be indicative of the mutation-inducing
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764 p53 Mutational Spectrum and Respiratory Tract Cancer
carcinogenic agents and may reflect underlying mutational
mechanisms. Therefore, the mutational spectra of the p53 gene
for lung cancer and for head and neck cancer may provide
information on the comparability of the underlying causes for
these cancers. In the present study, the p53 mutational spectra
(exons 5-8) were determined and compared for tumors of the
URT and LRT.
MATERIALS AND METHODS
The specimens were surgically resected, histologically con-
firmed tumors obtained from patients with non-small cell lung
cancer or head and neck cancer who were treated at the Uni-
versity of Pittsburgh Medical Center in conjunction with the
Pittsburgh Cancer Institute. Specimens for this study were se-
lected without regard to clinical stage, patient prognosis, therapy
regimen, metastasis, or primary origin of tumor site in the head
and neck region, except that tumors of the thyroid, esophagus,
and skin were not included. Cases were collected consecutively
and selection criteria were based on availability of tissue. Pa-
tients with prior treatment with radiation on chemotherapy were
not excluded from the study but comprised only a small portion
of the study population (15/95). Demographic information was
obtained by questionnaire at the time of tissue biopsy and was
entered into a computer data base by the personnel of the
Pittsburgh Cancer Institute Tissue and Serum Bank.
DNA was isolated from primary fresh-frozen tumor tissue
by guanidine thiocyanate extraction (33) using the commercially
available IsoQuick kit (MicroProbe, Garden Grove, CA). PCR
amplification and sequencing ofp53 exons 5 through 8 (includ-
ing exon-intron boundaries) were performed using previously
published PCR primers (34). All PCR reactions were carried out
in 100 pA total volume on an automatic thermocycler (Perkin
Elmer/Cetus 480 or 9600). The reaction conditions were: 10 msi
Tnis-HC1 (pH 8.3), 50 mM KC1, 1.5 mrvt MgCl2, 0.1% w/v
gelatin, 50 p.M each deoxynucleoside tniphosphate, 0.3 �.LM each
primer, and 1.25 units Taq polymerase (Perkin Elmer/Cetus,
Norwalk, CT). The PCR thermocycler parameters for all ampli-
fications consisted of an initial denaturation at 95#{176}Cfor S mm
followed by 28 cycles at 95#{176}Cfor 1 mm, 60#{176}Cfor 1 mm, and
72#{176}Cfor 1 mm. To confirm correct amplification, the products
were subjected to electrophoresis on 1.6% agarose minigels,
visualized by staining with ethidium bromide, and photographed
under UV light. Removal of primers and deoxynueleoside
tniphosphates as well as concentrations of the PCR products was
done by the use of Micron 100 microconcentrators (Amicon,
Beverly, MA). Direct sequencing of the double-stranded PCR
product was performed by a modification of the standard
dideoxynucleotide chain terminating method (35). PCR prod-
ucts were directly sequenced with fluorescent dye-labeled
dideoxynucleotides using Taq polymerase and cycle sequenc-
ing. The sequencing reactions were performed using the Taq
DyeDeoxy Terminator Cycle Sequencing kit (ABI, Foster City,
CA). Sequencing products were purified of unincorporated dye-
labeled dideoxynucleotides by processing through Centni-Sep
spin columns (Princeton Separations, Princeton, NJ). Sequenc-
ing products were electrophoretically fractionated through 6%
denaturing polyacrylamide gels (0.4-mm thick). Electrophore-
sis, band visualization, and sequence analysis were automati-
cally performed on the Applied Biosystems 373A automatic
sequencer. All mutations were confirmed by sequencing both
DNA strands.
RESULTS
Ninety-five tumor specimens were directly sequenced for
p53 mutations in exons 5-8. Mutations were identified in 29 of
these samples, for an overall frequency of approximately 31%.
All identified mutations were single-base substitutions. Four
were nonsense mutations leading to premature stop codons, 1
was a splice site mutation, 4 were same-sense mutations with no
change in amino acid, and 20 were missense mutations leading
to amino acid substitutions in the protein (Table 1). There was
a total of 16 base transitions (7 C-*T, 7 G->A, and 2 A-’G)
and 13 base transversions (8 G-�C, 3 G-+T, 1 C-�A, 1 C-+G).
The overall distribution of these mutations was relatively ran-
dom across p53 exons 5-8, with 8 mutations identified in exon
8, 3 in exon 7, 10 in exon 6, 7 in exon 5, and 1 splice site
mutation at the 3’ end of intron 4. The most frequently mutated
codons were codons 221, 273, and 299. The mutation at codon
273 was identified in three different URT tumor samples and
was always a G-*A transition at the second nucleotide position
of the codon. This missense mutation should lead to an amino
acid substitution of histidine for arginine in the protein. The
mutation at codon 221 was identified in four different tumor
samples (three URT and one lung) and was always the same
G-�C base substitution at the third nucleotide position of the
codon in the three lung tumors. This mutation leads to an amino
acid substitution of asparagine for glutamic acid in the protein.
The remaining codon 221 mutation was a G-�A base substitu-
tion at the third base position which leads to no amino acid
substitution. The codon 299 base substitution does not lead to an
amino acid substitution and is not located in one of the evolu-
tionanily conserved domains of p53. This silent mutation was
found in three different URT tumor specimens, was always a
G-#{247}Cbase substitution at the third nucleotide position of codon
299, and could not be identified in DNA extracted from the
nontumon tissue of patients. Most of the identified mutations
appeared to be hetenozygous, with both mutant and wild-type
peaks present on sequencing chrornatognaphs (Fig. 1). Tumor
specimen sectioning was performed with careful pathological
review but specimens were not microdissected. The true allelic
status of these mutations (homozygous or heterozygous) is
difficult to assess since normal tissue contamination of the gross
tumor specimen is unavoidable.
The mutational data was stratified for comparison between
the URT and LRT (Table 1). The frequency of identified mu-
tations was nearly identical at 30% (19/63) for URT tumors and
31% (10/32) for lung cancer. The functional nature of the
various types of mutations did not differ dramatically between
URT and LRT tumors (Fig. 2). Missense mutations made up a
significant majority of all identified mutations. Mutations lead-
ing to protein truncation (splice site and premature stop codons)
comprised approximately 16 and 20% of mutations identified in
URT and LRT tumors, respectively.
Differences in p53 mutational profiles were observed be-
tween URT and LRT tumors. Mutations identified in lung
cancer were primarily transversions (70%), whereas the muta-
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URT Lung
Sample Exon Codon Mutation” Amino acid Sample Exon Codon Mutation’� Amino acid
5CC 006 8 282 IGG -� IGG Ang -� Trp EDRN 4877 5 167 �AG -� TAG Gln -� stop
5CC 007 5 175 C�jC -� CAC Arg -� His EDRN 5106 6 224 GA�I -p GAl Glu -� Asp5CC 015 8 273 C�1T -� CAT Arg -p His EDRN 5357 6 192 �AG - lAG Gln -� stop
5CC 027 7 248 1�GG -p TGG Arg -� Trp EDRN 5749 8 282 IGG - IGG Arg -� Trp
5CC 037 8 273 C�1T -� CAT Arg -� His EDRN 5791 5 158 C�1C -� CIC Arg -� Leu
5CC 041 8 299 Cr�1 -� Cit Leu -� Leu EDRN 6218 6 189 .�1CC -� �CC Ala -p Pro5CC 042 7 249 AGG -� �1GG Mg -� Gly EDRN 6334 6 221 GA� -* GA� Glu -p Asp5CC 066 6 189 11CC -� �CC Ala -k Pro EDRN 6377 6 221 GA� -� GA� Glu -� AspSCC 071 5 132 MG -� A�G Lys -� Arg EDRN 6425 7 249 AG�1 -� AGI Arg -� 5cr5CC 072 5 179 LAT -� �AT His -� Asn EDRN 6886 6 221 GA�1 -� GA� Glu -� Asp
5CC 074 6 221 GA�1 -� GA� Glu -A Glu5CC 078 5 158 .CGC -� �iGC Arg -� Gly
EDRN 4141 Intron - GT -� AT Splice siteEDRN 4135 6 196 IGA -� IGA Arg -� stop
EDRN 4149 6 196 IGA -� IGA Arg -* stop
EDRN 4882 8 299 CT�j -� C1� Leu -� Leu
EDRN 5176 8 299 CF�j -� CT� Leu -� Leu
EDRN 5700 5 175 C�jC -� CAC Arg -� His
EDRN5811 8 273 C�T-ACAT Arg-*His
Total analyzed (n = 63) Total analyzed (n = 32)
Mutations (19/63) Mutations (10/32)Transitions (13/19) 7 G-*A: 4 C-�T: 2 A-aG Transitions (3/10) 3 C-*T
Transversions (6/19) 4 G-sC: 1 C-+A: 1 C-Ki Transversions (7/10) 3 G-T : 4 G-sC
U LRTU UR
at
U
a
I
Fig. 2 Comparison of the frequency of types ofp53 mutation observed
in URT and lung tumors.
type of mutation
Clinical Cancer Research 765
a Coding strand sequence.
Table 1 p53 Mutational Spectra for URT and LRT
C � I A I C N M A C. 1 C C. A A
Al’� IA
1\! \ � / \f‘�‘J� � t “ \ 1’_g�’�j� ‘
Fig. I p53 DNA sequence eleetrochromatogram of an URT tumor(EDRN 4135). Sequence around p53 codon 196 demonstrating both
wild-type (C) and mutated (7) alleles.
tions observed in URT tumors were primarily transitions (68%).
Comparison of the frequency of specific base substitutions
between URT tumors and lung cancer clearly demonstrates a
difference in observed mutational spectra (Fig. 3). Lung cancer-
associated mutations included 3 C-�T, 3 G-*T, and 4 G-�C
base substitutions (coding strand). URT tumor mutations in-
eluded 7 G-�A, 4 C-�T, 2 A-�G, 4 G-�C, 1 C-�A, and 1
C-�G base substitutions (coding strand). Mutations involving
G:C-’A:T transitions were the most frequently identified alter-ation in URT tumors (1 1/19) and G:C-�A:T mutations at CpG
sites, which are characteristic of endogenous processes, were
observed frequently in URT tumors (9/19) but only rarely in
lung tumors (1/10). Additionally, G:C-�T:A tnansversions,
which were relatively common in lung tumors (3/10), were rare
in URT tumors (1/19). No significant associations were ob-
served when the mutational data were further stratified to look
for possible associations between specific nucleotide substitu-
tions by tumor site and occupation (data not shown). Two-
variable analyses failed to show any statistical relationship
between the type of mutation and gender, smoking history, or
history of alcohol consumption (Table 2).
DISCUSSION
The combined mutational data on URT and LRT tumors
suggest that p53 mutations are relatively frequent in respiratory
tract cancers, and that the nature of the mutations and their
distribution are not unusual when compared with p53 mutations
identified in other human cancers. However, comparison of the
frequency of specific base substitutions between URT and LRT
tumors showed a striking difference in p53 mutational profiles.
Among tissues with mutations, transversion mutations occurred
twice as often in lung tumors and transition mutations twice as
often in URT tumors. This difference achieved a borderline
level of statistical significance (P = 0.064). This difference
could not be explained by differences in environmental expo-
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a UR
. LR
c->T - G->A � A->G � C->A C->G G->T G->Ctype of mutatIon
766 p53 Mutational Spectrum and Respiratory Tract Cancer
C0a
E0
UCa0�
Fig. 3 Specific p53 nucleotide substitutions observed in URT and lung
tumors (coding strand sequences).
sure as it relates to tobacco smoke, alcohol consumption, or
occupation.
Previous reports have found similar mutational profiles in
SCCHNs and squamous cell carcinoma of the lung (27, 29). The
difference observed in the p53 mutational spectra of URT and
LRT may be due, in pant, to the difference in the pathology of
the tumors. The majority of the URT tumors in which p.53 muta-
tions have been identified were squamous cell carcinomas (86%).
The two URT samples that were not squamous cell carcinomas
were 5CC 006 (adenoid cystic carcinoma) and 5CC 015 (muco-
epidermoid carcinoma), both of which displayed transition muta-
tions, whereas the majority of the lung cancer specimens (21/32)
and lung tumors identified with mutations (7/10) were primarily
adenocarcinornas (Table 3). The three lung tumors identified with
p53 mutations that were not adenocarcinomas were EDRN 4877,
EDRN 5791 (squamous cell carcinomas), and EDRN 6334 (large
cell carcinoma). However, the difference in pathology between
URT and lung tumors may not totally account for the observed
difference in mutational spectra, since comparison of base substi-
tutions identified in only the squamous cell carcinoma samples still
suggests a difference in the mutational profile between URT and
LRT tumors. One of the two lung squamous cell carcinoma sam-
ples identified with p53 mutations was a G:C-�T:A base substi-
tution, whereas this type of mutation was identified in only 1 of 17
URT squamous cell carcinoma mutations. Additionally, a recent
comparison of p53 mutations reported for adenocarcinoma and
squamous cell carcinoma of the lung show similar profiles with
approximately identical transversion frequencies (36).
An even more pronounced difference in the p53 mutational
spectra is observed between the URT and LRT when only
functionally relevant mutations are compared. Elimination of
the codon 299 (G-��C) same-sense mutation identified in three
URT tumors results in a URT mutational profile which consists
of 81% (13/16) base transitions, of which 85% (11/13) were
G:C-�A:T. A difference in mutational spectrum between head
and neck squamous cell carcinoma samples and lung cancer has
been previously reported (37). The p53 mutational profile of
URT tumors derived from our data more closely fits that of
previously reported p53 mutational spectra observed in gastro-
intestinal tract cancers (36, 38, 39). A recent, extensive compi-
lation and comparison of 2567 reported p53 mutations in human
Table 2 Com panison of m utation type t
drinking
o gender, smok ing, and
No mutation
(%)Transition
(%)Transversion
(%)x2
test
Gender
Men (54)
Women (41)
Smoking
Yes (80)
No (15)
Drinking
Yes (58)
No (33)
66.6
73.2
71.2
60.0
68.9
69.7
20.4
12.2
17.5
13.3
19.0
12.1
13.0
14.6
11.3
26.7
12.1
18.2
P = 0.57
P = 0.28
P = 0.75
cancers showed a similar, albeit not as pronounced, difference
between the profiles of 897 lung cancer mutations and 524 head
and neck tumor mutations (36). Approximately 1.5 times as
many transversions were observed in lung tumors as in head and
neck tumors with a preponderance of G:C-+T:A lung mutations
(40%) and a preponderance of G:C-�A:T (31%) head and neck
mutations (36). The reasons for the observed difference in
mutational profiles between the URT and LRT is uncertain, but
may reflect differences in both exogenously induced tissue-
specific mutational mechanisms and underlying endogenously
induced mutation rate variation between the tissues of the upper
and lower respiratory systems. The G:C-�A:T mutations at
CpG dinucleotides are due to frequent rnethylation of cytosine
to 5-methylcytosine and subsequent spontaneous deamination to
thyrnine, resulting in G:T mismatches which may not be me-
paired accurately (40). This mutational mechanism is an endo-
genous process for which no exogenous factors have yet been
identified that alter the frequency of methylation, deamination,
or the efficiency of repair leading to these CpG-related muta-
tions (41). In our study, G:C-�A:T mutations at CpG sites were
observed in 47% of URT tumors but only in 10% of lung
tumors. This suggests that endogenous biological factors may
play a role in determining some of the observed differences in
the p53 mutational spectra between the upper and lower respi-
ratory systems. The reasons for differences in endogenous tis-
sue-specific mutation rates for the p53 gene, as ascertained by
CpG mutations, is unknown but has been well documented
(reviewed in Ref. 41).
The higher frequency of G:C-#{247}T:A mutations in lung
tumors is representative of the type of mutation known to be
caused by polycyclic aromatic hydrocarbons, most notably ben-
zo(a)pyrene, that are present in tobacco smoke. The upper
aerodigestive and LRTs share tobacco smoke as a significant
common exogenous mutagenic risk factor. However, our data
suggest that either different tobacco-related mutagens or differ-
ent mutational mechanisms are involved in p53 mutation in
URT and LRT cancers. Tobacco smoke consists of many po-
tentially mutagenic substances, some of which have been well
characterized, such as N-nitrosamines and polycyclic aromatic
hydrocarbons, but tobacco smoke also consists of many other
compounds which have not been well defined and whose mu-
tagenic potentials are unknown. The concentration and duration
of exposure to the various compounds in tobacco smoke would
be expected to vary considerably between the URT and LRT.
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Clinical Cancer Research 767
Table 3 Sample pathology, mutational status, and smoking history
Code’� Path” p53C Smoke Code Path p53 Smoke Code Path p53 Smoke
003 5CC wt + 048 5CC wt + L4789 ADC wt +
004 5CC wt + 055 5CC wt + L4841 ADC wt +
006 ACC mu - 058 5CC wt + L4877 5CC mu +
007 SCC mu + 059 MVC wt + L4899 5CC wt +
009 5CC wt + 060 5CC wt + L4926 5CC wt +
010 5CC wt + 061 5CC wt + L5011 5CC wt +
01 1 5CC wt + 066 5CC mu + L5037 ADC wt +
012 5CC wt + 067 5CC wt + L5106 ADC mu +
013 5CC wt + 069 5CC wt + L5224 ADC wt -
015 MEC mu + 070 5CC wt + L5229 SCC wt +
017 5CC wt + 071 5CC mu + L5317 ADC wt -
019 5CC wt + 072 SCC mu + L5357 ADC mu +
020 5CC wt - 073 5CC wt + L5506 ADC wt -
024 5CC wt + 074 5CC mu - L5517 5CC wt +
026 5CC wt - 075 5CC wt + L5749 ADC mu +
027 5CC mu + 078 5CC mu - L5791 SCC mu +
030 5CC wt + 079 5CC wt + L6020 5CC wt +
031 SCC wt - 080 5CC wt + L6041 ADC wt +
033 5CC wt + 4135 5CC mu + L6087 5CC wt +
034 5CC wt - 4141 5CC mu + L6120 ADC wt -
035 MED wt + 4149 5CC mu + L6218 ADC mu +
036 5CC wt + 4174 5CC wt + L6311 ADC wt +
037 SCC mu + 4684 5CC wt + L6334 LCC mu +
038 5CC wt + 4882 5CC mu - L6377 ASC mu -
039 5CC wt + 5176 5CC mu + L6425 ADC mu +
040 5CC wt - 5328 5CC wt + L6272 ADC wt +
041 5CC mu + 5416 5CC wt + L6879 ADC wt +
042 5CC mu + 5624 5CC wt + L6886 ADC mu -
043 5CC wt + 5700 5CC mu - L691 1 ASC wt +
044 5CC wt + 5710 5CC wt + L6906 ADC wt +
045 5CC wt + 5811 5CC mu + L6957 ADC wt +
046 5CC wt + L4763 5CC wt +
a Lung sample codes are preceded by L.b 5CC, squamous cell carcinoma; ACC, adenoid cystic cancer; MEC, mucoepidermoid cancer; MED, mild epithelial dysplasia; MVC, mixed
verrueous cancer; ADC, adenocarcinoma; ASC, adenosquamous carcinoma; LCC, large cell carcinoma.
C mu, mutation; wt, wild type.
Additionally, the upper aerodigestive tract is exposed to differ- ACKNOWLEDGMENTS
ent known environmental risk factors not associated with the We thank Diana Kerestan and Anee Deka for their assistance in
LRT. Some of these risk factors include alcohol, sodium nitrites, obtaining DNA sequencing data, Christa Lese and Jaya Reddy for their
and smokeless tobacco products. The fact that the p53 muta- assistance in the acquisition of specimens and associated patient infor-
tional profile observed for URT tumors was more similar to that mation, and the staff of the Pittsburgh Cancer Institute Tissue and Serum
previously reported in gastric cancer than to LRT tumors would Bank for coordination of specimen accrual and processing.
suggest that these additional factors may play a role in deter-
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1995;1:763-768. Clin Cancer Res J C Law, T L Whiteside, S M Gollin, et al. upper and lower respiratory tract.Variation of p53 mutational spectra between carcinoma of the
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