Prognostic value of p53 mutations, bax, and spontaneous apoptosis in maxillary sinus squamous cell carcinoma.
ABSTRACT Many researchers have attempted to correlate p53 mutation and spontaneous apoptosis with the effectiveness of radiochemotherapy and with prognosis in several malignancies.
The current study group consisted of 70 Japanese patients with maxillary sinus squamous cell carcinoma (SCC). Fifty seven patients were treated with radiochemotherapy followed by total or partial maxillectomy, and the remaining 13 patients were treated with radiotherapy alone. Tumor biopsy specimens at pretreatment status were examined for apoptosis-related proteins such as p53 protein, Fas, bax, bcl-x, and apoptosis using immunohistologic methods. The proportion of apoptotic cells labeled by single stranded DNA antibody was expressed as an apoptotic index (AI). p53 mutations at exons 5 through 8 were analyzed by direct sequence on polymerase chain reaction amplified products obtained from laser microdissected tissues. The effectiveness of radiochemotherapy was investigated histologically on surgically dissected specimens.
p53 mutations were identified in 20 (29%) of 70 patients. p53 protein was overexpressed in 39 patients (56%), Fas in 20 patients (29%), bax in 40 patients (57%), and bcl-x in 33 patients (47%). Overexpression of bax was associated with negativity of bcl-x (P = 0.015) and with high AI (P = 0.024). Low AI and/or p53 mutation in the pretreatment tissues correlated with low histologic effectiveness of radiochemotherapy (P = 0.048, P = 0.019, respectively). Kaplan-Meier analysis as well as univariate analysis using the Cox proportional hazards model showed that low histologic effectiveness of radiochemotherapy (P = 0.0281, P = 0.0284, respectively), p53 mutations (P = 0.0095, P = 0.0187, respectively), negativity of bax (P = 0.0069, P = 0.0191, respectively), and low AI (P = 0.0134, P = 0.0407, respectively) were significantly related to worse disease-free survival. Multivariate analysis showed AI as an independent factor predicting for disease-free survival (P = 0.0455).
The p53 mutations, expression of bax, and levels of spontaneous apoptosis have prognostic value in maxillary sinus SCC; AI especially is an independent factor for disease-free survival. A high level of spontaneous apoptosis induced by overexpression of bax may increase sensitivity of radiochemotherapy resulting in good prognosis, while p53 mutation may lead to resistance against radiochemotherapy, resulting in poor prognosis.
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ABSTRACT: Nasal natural killer/T-cell lymphoma (NNKTL) is associated with the Epstein-Barr virus (EBV), and has distinct histological features such as angiocentric and polymorphous lymphoreticular infiltrates that contain too many cell types, including tumour and inflammatory cells. We have previously shown that intercellular adhesion molecule (ICAM)-1 is expressed in NNKTL cells, and that soluble ICAM-1 (sICAM-1) is significantly increased in patients' sera. However, the functional role of sICAM-1 remains unknown. In the present study, we found that EBV-positive NNKTL cell line SNK6 secreted sICAM-1 in a time-dependent manner. Moreover, exogenous sICAM-1 enhanced the growth of SNK6 cells in a dose-dependent manner. Comparatively, neutralising ICAM-1 and LFA-1 antibodies, as well as the LFA-1 blocker simvastatin, caused a dose-dependent reduction in the number of viable SNK6 cells. Double immunohistological staining of NNKTL tissues confirmed that CD56 positive lymphoma cells co-expressed LFA-1. Moreover, serum sICAM-1 levels in NNKTL patients decreased after treatment, suggesting that the levels reflected disease progression. We conclude that NNKTL cells secrete sICAM-1 that acts as an autocrine factor for lymphoma progression, and suggest that simvastatin may be a potential candidate to treat NNKTL.Experimental hematology 04/2013; · 3.11 Impact Factor
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ABSTRACT: Sinonasal carcinomas are rare tumours with an unfavourable prognosis whose management is difficult and complex, leading to high morbidity and mortality despite improvements in the field of surgery and radiotherapy. An elevated number of these tumours can be attributed to occupational exposure. In comparison with other head and neck malignancies, studies of molecular changes in these tumours are infrequent. This review was focused on findings about the epidemiology and molecular and phenotypic characterisation of sinonasal carcinomas, which can potentially be useful for diagnosis and treatment. The increasing knowledge about the molecular biology that underlies their carcinogenesis may help to identify precursor lesions, prognostic markers and markers that predict chemoradiotherapy response and, finally, to identify potential molecular targets that will expand treatment options.Acta Otorrinolaringológica Española 05/2012;
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ABSTRACT: The oxygen paradox tells us that oxygen is both necessary for aerobic life and toxic to all life forms. Reactive oxygen species (ROS) touch every biological and medical discipline, especially those involving proliferative status, supporting the idea that active oxygen may be increased in tumor cells. In fact, metabolism of oxygen and the resulting toxic byproducts can cause cancer and death. Efforts to counteract the damage caused by ROS are gaining acceptance as a basis for novel therapeutic approaches, and the field of prevention of cancer is experiencing an upsurge of interest in medically useful antioxidants. Apoptosis is an important means of regulating cell numbers in the developing cell system, but it is so important that it must be controlled. Normal cell death in homeostasis of multicellular organisms is mediated through tightly regulated apoptotic pathways that involve oxidative stress regulation. Defective signaling through these pathways can contribute to both unbalance in apoptosis and development of cancer. Finally, in this review, we discuss new knowledge about recent tools that provide powerful antioxidant strategies, and designing methods to deliver to target cells, in the prevention and treatment of cancer.Archives of Toxicology 07/2012; 86(11):1649-65. · 5.22 Impact Factor
Prognostic Value of p53 Mutations, Bax, and
Spontaneous Apoptosis in Maxillary Sinus Squamous
Nobuyuki Bandoh, M.D.
Tatsuya Hayashi, M.D., Ph.D.
Kan Kishibe, M.D., Ph.D.
Miki Takahara, M.D.
Masanobu Imada, M.D., Ph.D.
Satoshi Nonaka, M.D., Ph.D.
Yasuaki Harabuchi, M.D., Ph.D.
Department of Otolaryngology, Asahikawa Medical
College, Hokkaido, Japan.
Supported by a Grant-in-Aid (No.11470353) for
scientific research from the Ministry of Education,
Science and Culture, Japan.
The authors thank Yutaka Kohgo, M.D., Ph.D., of
the third Department of Internal Medicine, Asa-
hikawa Medical College, for the help in using the
laser capture microdissection system.
Address for reprints: Yasuaki Harabuchi, M.D.,
Ph.D., Department of Otolaryngology, Asahikawa
Medical College, Midorigaoka E 2-1-1-1, Asa-
hikawa, Hokkaido 078-8510, Japan; Fax: (011)
?81-166-68-2559; E-mail: hyasu@asahikawa-
Received October 3, 2000; revision received No-
vember 12, 2001; accepted December 3, 2001.
BACKGROUND. Many researchers have attempted to correlate p53 mutation and
spontaneous apoptosis with the effectiveness of radiochemotherapy and with
prognosis in several malignancies.
METHODS. The current study group consisted of 70 Japanese patients with maxil-
lary sinus squamous cell carcinoma (SCC). Fifty seven patients were treated with
radiochemotherapy followed by total or partial maxillectomy, and the remaining13
patients were treated with radiotherapy alone. Tumor biopsy specimens at pre-
treatment status were examined for apoptosis-related proteins such as p53 protein,
Fas, bax, bcl-x, and apoptosis using immunohistologic methods. The proportion of
apoptotic cells labeled by single stranded DNA antibody was expressed as an
apoptotic index (AI). p53 mutations at exons 5 through 8 were analyzed by direct
sequence on polymerase chain reaction amplified products obtained from laser
microdissected tissues. The effectiveness of radiochemotherapy was investigated
histologically on surgically dissected specimens.
RESULTS. p53 mutations were identified in 20 (29%) of 70 patients. p53 protein was
overexpressed in 39 patients (56%), Fas in 20 patients (29%), bax in 40 patients
(57%), and bcl-x in 33 patients (47%). Overexpression of bax was associated with
negativity of bcl-x (P ? 0.015) and with high AI (P ? 0.024). Low AI and/or p53
mutation in the pretreatment tissues correlated with low histologic effectiveness of
radiochemotherapy (P ? 0.048, P ? 0.019, respectively). Kaplan-Meier analysis as
well as univariate analysis using the Cox proportional hazards model showed that low
histologic effectiveness of radiochemotherapy (P ? 0.0281, P ? 0.0284, respectively),
p53 mutations (P ? 0.0095, P ? 0.0187, respectively), negativity of bax (P ? 0.0069, P
? 0.0191, respectively), and low AI (P ? 0.0134, P ? 0.0407, respectively) were
significantly related to worse disease-free survival. Multivariate analysis showed AI as
an independent factor predicting for disease-free survival (P ? 0.0455).
CONCLUSIONS. The p53 mutations, expression of bax, and levels of spontaneous
apoptosis have prognostic value in maxillary sinus SCC; AI especially is an inde-
pendent factor for disease-free survival. A high level of spontaneous apoptosis
induced by overexpression of bax may increase sensitivity of radiochemotherapy
resulting in good prognosis, while p53 mutation may lead to resistance against
radiochemotherapy. resulting in poor prognosis. Cancer 2002;94:1968–80.
© 2002 American Cancer Society.
KEYWORDS: maxillary sinus carcinoma, p53 mutation, Fas, bax, bcl-x, apoptosis
index, effectiveness of radiochemotherapy, prognostic factor.
studies have shown that environmental factors or inflammations such
axillary sinus carcinoma is a relatively rare cancer in western
countries, but it is not always rare in Japan.1,2Epidemiologic
© 2002 American Cancer Society
as chronic sinusitis or human papilloma virus infec-
tion3seem to be major etiologic factors in the induc-
tion of maxillary sinus carcinoma. Understanding the
relationship between biologic and clinical behaviors
may lead to improvements in predicting clinical out-
come, but such has not yet been performed in maxil-
lary sinus carcinoma.
Tumor progression is characterized by an imbal-
ance between cell proliferation and apoptosis.4–6One
of the tumor suppressor genes, p53, contributes to
tumor suppression through at least two mechanisms
in response to DNA damage, i.e., arrest of cell prolif-
eration and induction of apoptosis.7,8The p53-related
apoptotic cell death is downregulated by bcl-2 and
some other members of the bcl-2 family, while up-
regulated by bax expression.9,10Bax is a proapoptotic
protein that induces apoptosis. Its action is activated
by p53 protein and neutralized when it is het-
erodimerized with bcl-2 and some other members of
the bcl-2 family, such as bcl-x.11Therefore, bax might
be involved in the apoptotic elimination of tumor cells
after exposure to DNA damaging drugs or radiation.
Aside from the p53-related passway, apoptosis is reg-
ulated by Fas 48-kD type I membrane protein of the
tumor necrosis factor receptor family.12The ligation of
Fas ligand and Fas stimulates an intracellular cascade
of events that leads to the induction of apoptosis in
cells, including cancer cells.13
Loss of the function of the p53 gene due to mu-
tation represents the most common genetic event
known in human cancer.14Alterations of the p53 gene
can be detected by either altered protein overexpres-
sion (immunohistology) or direct sequencing. Dereg-
ulation of the apoptotic pathway due to loss of p53
function probably contributes to treatment failure in
patients with cancer by making malignant cells more
resistant or sensitive to radiotherapy and chemother-
apy.9,15–17In head and neck squamous cell carcinoma
(SCC), a number of reports have documented overex-
pression of p53 protein18–21and p53 gene muta-
tion.16,19,22–25However, conflicting results have been
presented with regard to the correlation between
prognosis and overexpression of p53. Some studies
showed no correlation,19,26–28whereas others showed
a significant correlation between p53 expression and
prognosis.21,29,30Conversely, a majority of studies
showed an association of p53 mutation with poor
prognosis,20,31but some studies showed that there is
no association between p53 mutation and prognosis.19
Overexpression of bcl-x has been reported to pro-
tect tumor cells from apoptosis,32whereas increased
bax expression promotes apoptosis induced by cyto-
toxic drugs and radiation.33,34Expression of apoptosis-
related proteins such as bax,35–37bcl-x,38Fas,13and
spontaneous apoptosis39has been analyzed immuno-
histologically in head and neck SCC. However, the
results have not been uniform with regard to the cor-
sis.6,38–41Furthermore, to our knowledge, there is no
information with regard to primary maxillary sinus
The purpose of the current study was to identify
mutations in the p53 tumor suppressor gene, to eval-
uate the expression of apoptosis-related proteins such
as p53, Fas, bax, and bcl-x, and to examine the inci-
dence of apoptotic cells in maxillary sinus SCC. The
goal of the current study was to investigate correla-
tions among p53 mutation and the expression of ap-
optosis-related proteins and apoptosis, and to deter-
radiochemotherapy and prognosis.
PATIENTS AND METHODS
The study group consisted of 70 Japanese patients (49
male, 21 female) with a median age of 67 years (range,
36 to 86 years) who were treated for maxillary sinus
SCC in the Department of Otolaryngology, Asahikawa
Medical College between 1980 and 2000. Patient clin-
ical features are listed in Table 1. According to the
1997 International Union Against Cancer TNM staging
system (identical to the 1997 American Joint Commit-
tee on Cancer classification), T1 was not seen; T2 was
present in 8 patients (11%), T3 was present in 33
patients (47%), and T4 was present in 29 patients
(42%). Seven patients (10%) had lymph node metas-
Clinical Features of 70 Patients with Maxillary Sinus Squamous
Variable No. of patients (%)
TNM staging was performed according to the 1997 International Union Against Cancer system.
p53 and Apoptosis in Maxillary Sinus Carcinoma/Bandoh et al. 1969
tasis (N1 in all cases) at diagnosis. The classification of
tumor differentiation was well differentiated in 29 pa-
tients (42%), moderately in 24 patients (34%), and
poorly in17 patients (24%). Clinical data have in part
been reported elsewhere.42
Of 70 patients, 57 were treated with preoperative
radiochemotherapy followed by total or partial maxil-
lectomy. The preoperative radiochemotherapy in-
cluded local irradiation with a total dose of 50 Gy (2.5
Gy by; 20 fractions, 5 days a week) along with con-
comitant intramaxillary arterial infusion of 5-fluorou-
racil with a total dose of 5000 mg (250 mg by 20
times).42The remaining 13 patients were treated with
radiotherapy alone because of inoperable intracranial
invasion, advanced age with poor performance status,
refusal of surgery, or concomitant intercurrent illness.
The followup period ranged from 2 to 189 months,
with a median of 61 months for all patients and of 116
months for surviving patients.
The formalin fixed and paraffin embedded specimens
were obtained from transoral biopsy at the pretreat-
ment period. The specimens were cut into 5 ?m sec-
tions. The slides were deparaffinized in xylene and
ethanol. These sections were then incubated with 3%
hydrogen peroxidase for 30 minutes. The following
antibodies were used: anti-p53 monoclonal antibody
(clone DO-7, diluted 1:50; DAKO A/S, Glostrup, Den-
mark); anti- Fas monoclonal antibody (Apo-1, diluted
1:10; DAKO, Carpinteria, CA); anti-bax polyclonal an-
tibody (diluted 1:50; DAKO); anti-bcl-x polyclonal an-
tibody (diluted 1:50; DAKO); and anti-single stranded
(ss) DNA polyclonal antibody (diluted 1:200; DAKO
Japan, Kyoto, Japan) for the detection of apoptotic
cells. The slides for p53, bax, and bcl-x were placed in
10 mM citric acid buffer at pH 6.0, and the slides
underwent antigen retrieval for a 10 minute cycle at
750 W and 95 °C in a microwave oven.43The slides for
Fas underwent proteolytic digestion using proteinase
K (DAKO) for 5 minutes. These primary antibodies
were incubated overnight at 4 °C, followed by peroxi-
dase labeled dextran polymer (EnVision?, DAKO) for
30 minutes at room temperature. The slides were vi-
sualized by immersion in freshly prepared 0.02% dia-
minobenzidine solution for 10 minutes. The sections
were finally counterstained with Lillie-Mayer’s hema-
toxylin and mounted.
The sections were examined microscopically by
three of the authors (N.B., M.T., Y.H.) who had no
knowledge of patients or clinical outcome. The over-
expression of p53 was noted when more than 10% of
tumor nuclei were stained, as previously reported.44
The positive staining with Fas was defined when
membrane-staining tumor cells were found, as previ-
ously reported.13The expression of bax and bcl-x was
categorized as follows: negative if no staining was seen
in tumor cells or if only a weak positive and hetero-
geneous staining was observed in less than 30% of
tumor cells, and positive if staining was observed in
more than 30% of tumor cells, according to re-
ports.38,45More than 5000 tumor cells were counted to
calculate the apoptotic index (AI) in at least five high
power fields, and the AI values were defined as the
number of ss DNA positive cells per 1000 tumor
cells.39,40Sections with anti-mouse immunoglobulin
G1 monoclonal antibody were used as negative con-
trols. Sections of laryngeal carcinoma known to ex-
press immunohistologically detectable p53 served as
positive controls for p53 antibodies. Sections of tonsil
served as positive controls for Fas, bax, bcl-x, and ss
DNA Extraction, Polymerase Chain Reaction,
and DNA Sequences
The laser capture microdissection (LCM) system (Arc-
turus Engineering, Mountain View, CA) was used to
extract DNA from tumor cells, as previously reported.46
We used unmounted 5 ?m sections stained with he-
matoxylin and eosin (HE). A capture cap (Arcturus
Engineering) was then placed on the section compris-
ing tumor nests. On the use of a laser beam 30 ?m in
diameter, cells in 100 areas were transferred to the
cap, yielding 500-1000 cells per sample. The captured
cells were placed in a microtube containing digestion
buffer (50 ?L buffer containing 0.04% proteinase K, 10
mM Tris-HCL [pH 8.0], 1 mM ethylenediaminetet-
raacetic acid, and 1% polyoxyethlene sorbitan mono-
laurate [Tween 20]). The tube was equilibrated over-
night at 37 °C. The tube was then heated to 95 °C for 8
minutes to inactivate the proteinase K. It was used
directly as template for polymerase chain reaction
We performed PCR in a 30 ?L reaction mixture
containing 14.85 ?L H2O, 3 ?L PCR buffer (15mM
MgCl2, 10-fold concentrated), 3 ?L deoxynucleosid
triphosphate (2mM), 3 ?L primer 1 (2mM), 3 ?L
primer 2 (2mM), 0.15 ?L Taq DNA polymerase (Am-
pliTaq Gold, 5U/ ?L; Applied Biosystems, Foster City,
CA), and 3 ?L DNA (100ng/?L). Thirty eight cycles of
denaturation (94 °C, 1 minute), annealing (58-60 °C, 1
minute) and extension (72 °C, 1 minute) were carried
out in a DNA thermal cycler (Programmable Thermal
Controller PTC-100, MJ Research, Inc., Tokyo, Japan).
Oligomer primers were synthesized originally for ex-
ons 5-8 in the p53 gene (SIGMA GENOSIS, Ishikari,
Japan). The primer sequences are shown in Table 2.
Polymerase chain reaction products were purified us-
1970 CANCER April 1, 2002 / Volume 94 / Number 7
ing QIAquick PCR purification Kit (QIAGEN Inc.,
Chastworth, CA). Sequencing was performed in both
directions using the PCR primers detailed above. In all
cases, the DNA was double-stranded sequenced using
the ABI PRISM BigDye terminator cycle sequencing
ready reaction kit (Applied Biosystems). Subsequent
electrophoresis was performed using an ABI PRISM
377 DNA sequencer (Applied Biosystems) and the data
collected were analyzed using Applied Biosystems se-
quencing analysis software.
Histologic Effectiveness of Radiochemotherapy
Surgically resected specimens obtained from 57 pa-
tients with radiochemotherapy followed by total or
partial maxillectomy were examined. They were fixed
in 10% formalin, embedded in paraffin, and stained
with HE for histologic studies. The effectiveness of
radiochemotherapy was assessed according to the
evaluation system proposed by Shimosato et al.47as
follows: Grade 0, no characteristic change in tumor
cells and tumor structures; Grade I, characteristic
changes in tumor cells, but tumor structures have not
been destroyed; Grade II, in addition to characteristic
cellular changes, tumor structures have been de-
stroyed as a result of the disappearance of tumor cells,
with a variable number of viable tumor cells still re-
maining; Grade III, markedly altered and presumably
nonviable tumor cells are present singly or in small
clusters, and viable tumor cells are hardly seen; Grade
IV, no tumor cells in any section.
Two group comparisons were performed using the
Mann-Whitney U test, chi-square test, or Fisher exact
test and were summarized with their appropriate P
value. The Spearman regression coefficient was used
to examine the magnitude of selected association.
Overall survival time was measured from the date of
diagnosis to the date of death or last followup visit.
Disease-free survival time was measured from the
date of surgical removal of tumor to the date of first
relapse or to the date of last followup. The probability
of overall and disease-free survival was calculated us-
ing the Kaplan-Meier method and compared using the
log rank test. For determination of factors related to
overall survival and disease-free survival, a Cox pro-
portional hazards model was used. The final results of
these analyses are hazard ratios, 95% confidence in-
tervals, and Pvalue. A P value less than 0.05 was con-
sidered statistically significant.
Expressions of Apoptosis Related Proteins and Apoptosis
p53 was overexpressed in 39 (56%) of 70 patients, Fas
was overexpressed in 20 patients (29%), bax was over-
expressed in 40 patients (57%), and bcl-x was overex-
pressed in 33 patients (47%) (Fig. 1). Apoptotic cells
were identified in all biopsy specimens at pretreat-
ment, and the AI ranged from 0.2 to 7.2 with median
value of 1.8 (Fig. 1). The AI in patients with bax-
overexpression was significantly higher than that in
patients without bax (1.6 vs. 2.7, P ? 0.024; Fig. 2), but
no significant correlation was found between AI and
the other apoptosis-related proteins. Overexpression
of p53 did not correlate with expression of bax, bcl-x,
or Fas. Conversely, the expression of bax was associ-
ated with negativity of bcl-x (P ? 0.015, Table 3). There
was no correlation between apoptosis-related param-
eters and either age, tumor extension, lymph nodes, or
Mutation Analysis of the p53 Gene
Twenty mutations (29%) were identified in 70 patients
(Table 4). Eight mutations (40%) were identified in
exon 5, 2 (8%) in exon 6, 3 (12%) in exon 7, and 7 (35%)
in exon 8. Nineteen patients (95%) had a single amino
acid mutation, and one (5%) had a nonsense muta-
tion. Of 20 mutations, 16 (80%) were transition type,
including 10 (50%) CpG site G:C to A:T changes, 2
(10%) non-CpG site G:C to A:T changes, and 4 (20%)
A:T to G:C changes. The remaining four mutations
were transversion type, including two (10%) G:C to
C:G changes, one (5%) G:C to T:A change, and one
(5%) A:T to T:A change.
Sixteen (80%) of 20 patients with p53 mutation
showed overexpression of p53 protein, while 23 (46%)
of 50 patients without p53 mutation showed such
overexpression (P ? 0.016, Table 3). No correlation
was found between p53 mutation and age, tumor ex-
tension, lymph nodes, tumor differentiation, or ex-
pression of apoptosis-related parameters.
Sequences of Genomic Primers with Expected Size of PCR Products
Size of PCR
exon 5A5?-CTT CCT GCA GTA CTC CCC TG-3?
5?-CTC ACA ACC TCC GTC ATG TG-3?
5?-CGC CAT GGC CAT CTA CAA G-3?
5?-GCC CCA GCT GCT CAC CAT C-3?
5?-GCC TCT GAT TCC TCA CTG AT-3?
5?-CTG AGG TCT GGT TTG CAA CT-3?
5?-TTG TCT CCT AGG TTG GCT CTG-3?
5?-GGC TCC TGA CCT GGA GTC TT-3?
5?-TTC CTT ACT GCC TCT TGC TT-3?
5?-TGA ATC TGA GGC ATA ACT GC-3?
exon 5B 101 bp
exon 6 157 bp
exon 7131 bp
exon 8 239 bp
PCR: polymerase chain reaction.
p53 and Apoptosis in Maxillary Sinus Carcinoma/Bandoh et al.1971
Histologic Effectiveness of Radiochemotherapy
Surgical specimens obtained from 57 patients with
preoperative radiochemotherapy followed by total or
partial maxillectomy were investigated for histologic
effectiveness of radiochemotherapy. According to Shi-
mosato’s classification,47Grade II was found in 21
patients (37%), Grade III in 20 (35%), and Grade IV in
16 (28%). Neither Grade 0 nor Grade I were identified.
As shown in Table 5, only 1 (6%) of 16 patients
with p53 mutation in pretreatment tissues had no
tumor cells in the surgically resected tissues (Grade
IV), while 15 (37%) of 41 patients without p53 muta-
tion did (P ? 0.019). On the other hand, the AI in
pretreatment tissues positively correlated with histo-
logic effectiveness of radiochemotherapy (r ? 0.451, P
? 0.048, Fig. 3). No significant correlation was found
therapy and expression of p53, bax, bcl-x, or Fas.
FIGURE1. Representative immunohistologic features of a) p53, b) Fas, c) bax, d) bcl-x, and e) apoptotic cells in maxillary sinus squamous cell carcinoma (?200).
a) p53 protein shows nuclear staining, b) Fas shows membrane staining, and c and d) bax and bcl-x show cytoplasmic staining. e) Apoptotic cells (arrow) were
determined by their condensed appearance with small, fragmented nuclei stained by anti-single stranded DNA antibody (?400).
1972 CANCER April 1, 2002 / Volume 94 / Number 7