Prognostic value of the preserved expression of the E-cadherin and catenin families of adhesion molecules and of β-catenin mutations in synovial sarcoma

Original Paper

Prognostic value of the preserved expression of theE-cadherin and catenin families of adhesion moleculesand of b-catenin mutations in synovial sarcoma

Tsuyoshi Saito1, Yoshinao Oda1, Akio Sakamoto1, Sadafumi Tamiya1, Naoko Kinukawa2, Kenshi Hayashi3,

Yukihide Iwamoto4 and Masazumi Tsuneyoshi1*1 Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan2 Department of Medical Information Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan3 Division of Genome Analysis, Institute of Genetic Information, Kyushu University, Fukuoka, Japan4 Department of Orthopaedic Surgery, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

*Correspondence to:Masazumi Tsuneyoshi, MD,Department of AnatomicPathology (Second Department ofPathology), Pathological Sciences,Graduate School of MedicalSciences, Kyushu University,Maidashi 3-1-1, Higashi-ku,Fukuoka 812-8582, Japan

Received: 28 September 1999

Revised: 13 March 2000

Accepted: 2 May 2000

Published online:

21 September 2000

Abstract

This study addresses the immunohistochemical expression of the E-cadherin and catenin families

and mutations of the b-catenin gene detected by PCR±SSCP in synovial sarcoma. Immunohis-

tochemical analysis was performed for 72 cases, with follow-up data available on 62. The

prognostic value of the expression of these proteins was evaluated. Reduced immunoreactivity for

E-cadherin and a-catenin was signi®cantly correlated with a poor survival rate ( p=0.0040 and

0.0053, respectively). According to multivariate analysis, low AJC stage (stages I and II:

p<0.0001), the preservation of a-catenin expression ( p=0.0001), and a low necrotic rate

(<50%: p=0.0139) were independent favourable prognostic factors. Widespread aberrant

staining of b-catenin protein within cytoplasm and/or nuclei was observed in 28 cases (38.9%)

and was signi®cantly correlated with poor survival (p=0.0122). In addition, there was a trend

towards a correlation between widespread aberrant staining of b-catenin and the MIB-1 labelling

index ( p=0.0535). Mutational analysis of exon 3 of the b-catenin gene was performed for 49

cases. Nucleotide sequencing analysis revealed that four (8.2%) contained point mutations (three

in codon 32, GAC to TAC; one in codon 37, TCT to TTT). Survival data were available for three

out of four cases with b-catenin mutations; two of these patients died within 1 year (died of disease

at 6 and 11 months, respectively). These results suggest that E-cadherin and a-catenin undertake

important roles as intercellular adhesion molecules; their preserved expression is associated with a

better overall survival rate in synovial sarcoma and may have prognostic value. Abnormal levels of

b-catenin, with or without mutation, could contribute to the development and progression of

synovial sarcoma, through increasing the proliferative activity of the tumour cells. Copyright #2000 John Wiley & Sons, Ltd.

Keywords: synovial sarcoma; intercellular adhesion molecules; b-catenin; mutation; cell

proliferation; prognosis; immunohistochemistry; PCR±SSCP

Introduction

Recent reports have demonstrated that E-cadherin andits cytoplasmic binding proteins, the catenins (a-, b-,c-), are essential for intercellular junctions; theirdecreased expression corresponds to the poor prog-nosis associated with the dissemination of tumour cellsand the formation of metastases [1±7]. Furthermore, ithas been reported that b-catenin is a multifunctionalprotein which is also involved in the wingless/Wntsignal transduction pathway [8±11], in addition tobeing a cell±cell adhesion regulator when binding tothe carboxy-terminus of calcium-dependent transmem-brane E-cadherin adhesion molecules. The binding ofb-catenin to adenomatous polyposis coli (APC) proteinrequires phosphorylation of b-catenin by GSK (glyco-gen synthase kinase)-3b on serine/threonine residues,all of which are encoded in exon 3 of the gene. Incolorectal cancers, mutations of APC or b-cateninresult in the stabilization of b-catenin and a signi®cant

accumulation of this protein within the cytoplasm.Furthermore, increased b-catenin may translocate tothe nuclei, and could serve as a transcriptional factorby binding to the T-cell factor/lymphoid enhancingfactor (Tcf±Lef) family. In the APC±b-catenin±Tcfpathway, accumulated free cytoplasmic b-cateninbehaves as an oncoprotein. In fact, in addition toAPC alterations [9,12], mutations of b-catenin havebeen identi®ed in various cancers, such as colon cancerwith a normal APC gene [10,13], melanoma cell lines[14], ovarian cancer [15], endometrial carcinoma[16,17], medulloblastoma [18], prostate cancer [19],hepatocellular carcinoma [20,21], human gastric cancercell lines [22], human colorectal cancer cell lines [23],and childhood hepatoblastoma [24].

More recently, such mutations have also beenreported in bone and soft tissue tumours [25], but therelationship between the expression of adhesion mole-cules and prognosis, and the occurrence of mutationsof the b-catenin gene have not been reported for soft

Journal of PathologyJ Pathol 2000; 192: 342±350.DOI: 10.1002 /1096-9896(2000)9999 :9999<: :AID-PATH705>3.0.CO;2-R

Copyright # 2000 John Wiley & Sons, Ltd.

tissue sarcomas. The purpose of this study was toinvestigate whether the expression of adhesion mole-cules could help to predict prognosis, and to elucidatewhether not only b-catenin mutation, but also b-catenin accumulation within the cytoplasm and/ornuclei could contribute to cell proliferation and thedevelopment of synovial sarcoma. We examined theimmunohistochemical expression of E-cadherin andcatenins, and we investigated the occurrence of muta-tions of the b-catenin gene by polymerase chainreaction±single-strand conformation polymorphism(PCR±SSCP) in a large series of cases of synovialsarcoma.

Materials and methods

One hundred cases of synovial sarcoma were selectedfrom more than 1500 soft tissue sarcomas registered inthe Department of Anatomic Pathology, GraduateSchool of Medical Sciences, Kyushu University, Japanfrom 1955 to 1998; formalin-®xed, paraf®n-embeddedtissue from 72 cases was available for immunohisto-chemical study. There were 57 cases of monophasictype and 15 of biphasic type, de®ned as those in whichapparent glandular structures were recognized. Clinicaldata for these cases were collected from the medicalrecords. Survival data were available for 62 cases, withfollow-up from 1 to 232 months (mean 55.5 months).There was no statistically signi®cant difference in thesurvival rate between earlier cases (21 cases before1980) and later cases (41 cases after 1980) ( p=0.438).

We assessed correlations between the expression ofE-cadherin and catenins and the following clinico-pathological factors: sex, age, anatomical site, tumourdepth, histological subtype, the presence of rhabdoidcells, mitotic rate, nuclear grade, tumour necrosis, thenumber of mast cells, tumour size, and tumour stageaccording to the American Joint Committee (AJC)[26]. The in¯uence of these factors on prognosis hasbeen previously reported [27]. As for histological type,`high glandular' implies the biphasic type, whereas `lowglandular' includes the monophasic type. Tumournecrosis was measured in the resected tumour or inall the slides available. Grade I nuclei showed regularborders and an even distribution of chromatin, whileGrade III nuclei had a vesicular chromatin pattern andprominent nucleoli; grade II nuclei were of intermedi-ate morphology.

Immunohistochemistry

Immunohistochemistry was used for the analysis of E-cadherin, a-catenin, b-catenin, and c-catenin proteinexpression. After antigen retrieval by a microwaveoven technique, the sections were incubated at 4uCovernight with monoclonal antibodies to E-cadherin,a-catenin, b-catenin, and c-catenin (all supplied byTransduction Laboratories) diluted at 1 : 1000, 1 : 100,1 : 200, and 1 : 200, respectively, followed by the use ofa streptavidin±biotin±peroxidase kit (Nichirei, Tokyo,

Japan) and haematoxylin counterstaining. Tissue fromnormal human gastric mucosa was used as a positivecontrol. Immunohistochemistry was also performedwith anti-Ki-67 monoclonal antibody (MIB-1, 1 : 100;Immunotech, Marseille, France) as an indicator of cellproliferation. The MIB-1 labelling index (LI) wasexpressed as the percentage of Ki-67-positive cellsbased on a count of 1000 tumour cells.

Evaluation of staining

Staining intensity was graded using a method pre-viously utilized [8,28]. Cells staining less intensely thanthose in the positive control were de®ned as negative,and the section was then scored semiquantitativelyaccording to the proportion of positive cells: +, morethan 90% positive; +/x, between 10% and 90%positive; x, less than 10% positive. Sections showingmore than 90% of positive cells were described ashaving `preserved type' expression, while other cate-gories were described as `reduced type'. Cellularlocalization of staining was also noted. Immuno-reactivity for b-catenin was described as positive onlywhen localized at the cellular membrane, whereasintracytoplasmic or aberrant nuclear expression wasdescribed as negative. In biphasic types of synovialsarcoma, immunoreactivity was evaluated in both the®brous and the glandular components, and if at leastone component was described as preserved type, thenthat case was classi®ed accordingly. As a recent reporthas described [11], sections with aberrant expression ofb-catenin protein in more than 75% of tumour cellswere judged as showing widespread aberrant staining.

Statistical analysis

Follow-up data were available in 62 cases. Thesigni®cance of the expression of E-cadherin andcatenins on the overall survival rate was estimatedusing the log-rank test (p<0.05 considered signi®cant).For those factors showing a signi®cant in¯uence onsurvival, Kaplan±Meier plots were constructed. ACox's proportional hazards model was ®tted to thedata in order to assess independent prognostic factors.

Polymerase chain reaction±single-strandconformation polymorphism (PCR±SSCP) andmutational analysis of b-catenin

Genomic DNA was puri®ed from 64 cases of synovialsarcoma. The tissue had been ®xed in 10% formalde-hyde and embedded in paraf®n wax. Standard protei-nase K digestion and phenol/chloroform extractionwere employed. A genomic PCR fragment includingexon 3, corresponding to the serine/threonine residues,which are essential phosphorylation sites for GSK-3b,was ampli®ed using the primers 5k-AAA GCG GCTGTT AGT CAC TGG-3k (sense) and 3k-GAC TTGGGA GGT ATC CAC ATC C-5k (antisense) [19]. PCRwas carried out in a Gene Amp PCR System 9600(Perkin Elmer, Foster City, CA, USA). Human

E-cadherin and catenins in synovial sarcoma 343

Copyright # 2000 John Wiley & Sons, Ltd. J Pathol 2000; 192: 342±350.

genomic DNA (CLONTECH) was used as a positivecontrol for each PCR and the subsequent reactions.We also con®rmed that there was no contamination ineach PCR and the subsequent reactions by usingdistilled water instead of template DNA. SSCP wasperformed using a gel containing 12.5% acrylamide(GenePhor2, Amersham Pharmacia Biotech, Uppsala,Sweden) and a DNA fragment analyzer (GenePhor2,Amersham Pharmacia Biotech, Uppsala, Sweden) andthen visualized by a DNA silver staining kit (Gene-Phor2, Amersham Pharmacia Biotech).

DNA sequencing

To increase the quantity of mutant DNA prior tosequencing, not only extra bands which seemed to beaberrantly migrating, but also the major bands wereexcised from the SSCP gel and reampli®ed and thensequencing was performed. The sequence data werecollected by ABI Prism 310 Collection Software andwere analyzed by Sequencing Analysis and SequenceNavigator Software (Perkin Elmer).

Results

Immunohistochemical ®ndings

The results of the immunohistochemical study aresummarized in Table 1 (Figure 1). In the biphasic typeof synovial sarcoma, heterogeneous immunohisto-chemical patterns for E-cadherin and catenins werefrequently seen together in the same tumour, withintense immunoreactivity in the glandular components,but weak or only faint immunoreactivity in the ®brouscomponents. As a result, no case showed preservedimmunoreactivity for E-cadherin and catenins in the®brous components of the biphasic type of synovialsarcoma. The preserved expression for E-cadherin anda-catenin was signi®cantly correlated with increasedsurvival (log-rank test: p=0.0040 and p=0.0053,respectively: Figures 2A and 2B). Although there wasno statistical signi®cance between preserved immuno-reactivity for b-catenin or c-catenin and prognosis,those groups demonstrated a tendency for long-termsurvival, compared with groups which showed reducedexpression (p=0.13 and p=0.15, respectively). Therewere signi®cant correlations between preservedimmunoreactivity for E-cadherin and catenins (data

not shown). The groups with preserved co-expression

of E-cadherin and a-catenin had a much better

prognosis than those with preserved expression of E-

cadherin only ( p=0.0313, Figure 2C). Furthermore,

within the monophasic type of synovial sarcoma,

preserved immunoreactivity for E-cadherin was also

signi®cantly correlated with a favourable survival rate

(p=0.0487, Figure 2D). Immunohistochemical analy-

sis of the accumulation of b-catenin within tumour

cells was also carried out. The aberrant expression of

b-catenin protein within the cytoplasm and/or nuclei

was observed to a variable degree in 42 cases (58.3%,

Figure 3). Amongst these, widespread aberrant staining

was observed in 28 (38.9%), which signi®cantly affec-

ted the overall survival rate (p=0.0122, Figure 2E)

and which showed a limited correlation with MIB-1 LI

(p=0.0535, Table 2). As a previous report [8] has

described, cytoplasmic expression of E-cadherin was

also observed in 21 of 72 cases, in all of which the

staining was of reduced type. There was no signi®cant

correlation between prognosis ( p=0.297) or MIB-1 LI

(p=0.801) and aberrant expression of E-cadherin.

Although this phenomenon was not seen with a-

catenin staining, there was a signi®cant correlation

between preserved a-catenin expression and MIB-1 LI

(Table 3: p=0.0046).

Table 1. Immunohistochemical results

Monophasic synovial sarcoma (57 cases) Biphasic synovial sarcoma (15 cases)

Preserved type

Reduced type

Preserved type

Reduced type

(+) (+x) (x) (+) (+x) (x)

E-cadherin 7 10 40 13 0 2

a-catenin 3 6 48 9 1 5

b-catenin 5 6 46 10 1 4

c-catenin 1 7 49 8 3 4

Table 2. Correlation between widespread aberrant stain-ing of b-catenin and MIB-1 LI

Widespread stainingof b-catenin

MIB-1 LI

Average SD

(x) n=44 10.076 8.038(+) n=28 14.132 9.289

p=0.0535

Table 3. Correlation between expression of a-cateninand MIB-1 LI

Expression of

a-catenin

MIB-1 LI

Average SD

(x) n=60 12.931 8.909

(+) n=12 5.264 3.282

p=0.0046

344 T. Saito et al.


Correlation between clinicopathologicalparameters and the expression of E-cadherin andcatenins

The relationships between clinicopathological param-

eters and the expression of E-cadherin and catenins are

summarized in Table 4. There were strong relationships

between high glandularity and E-cadherin and catenin

expression (p<0.0001). There were also signi®cant

relationships between the expression of E-cadherin

and the absence of rhabdoid cells, low mitotic rate(<15 per 10 HPF), and low nuclear grade (grade I or

II) (p<0.05). Among the clinicopathological param-

eters and the expression of E-cadherin and catenins in-

cluding aberrant b-catenin staining and MIB-1 LI usingstepwise multivariate survival analysis with Cox's pro-

portional hazards model, a low AJC stage ( p<0.0001),

preserved a-catenin expression ( p=0.0001), and a low

necrotic rate (p=0.0139) were con®rmed as indepen-dent favourable prognostic factors (Table 5).

A B

DC

FE

Figure 1. (A±D) Preserved immunoreactivity for E-cadherin (A), a-catenin (B), b-catenin (C), and c-catenin (D) is observed in theglandular components of biphasic synovial sarcoma. (E, F) In a case of monophasic ®brous synovial sarcoma, spindle-shaped®broblastic tumour cells are positive for E-cadherin (F)



b-catenin mutation

We were able successfully to amplify and screenmutations of exon 3 of the b-catenin gene by PCR in

49 out of 64 DNA samples; aberrantly migrating bandswith b-catenin mutations were observed in four casesby PCR±SSCP analysis (Figure 4). Nucleotide sequen-cing analyses showed that these four tumours con-

Figure 2. (A) Survival curve of patients with synovial sarcoma according to E-cadherin expression. Survival with preserved E-cadherin expression in tumour cells was signi®cantly better than with reduced expression (log-rank test: p=0.0040). (B) Survivalcurve of patients with synovial sarcoma according to a-catenin expression. Survival with preserved a-catenin expression wassigni®cantly greater than with reduced expression (log-rank test: p=0.0053). (C) Survival curve of patients with synovial sarcomaaccording to co-expression of E-cadherin and a-catenin. Survival with preserved co-expression of E-cadherin and a-catenin wassigni®cantly longer than with preserved expression for E-cadherin only (log-rank test: p=0.0313). (D) Survival curve of patients withmonophasic synovial sarcoma according to E-cadherin expression. Survival with preserved expression of E-cadherin was signi®cantlybetter than with reduced expression (log-rank test: p=0.0487). (E) Survival curve of patients with synovial sarcoma according towidespread aberrant staining of b-catenin. Survival with aberrant staining of b-catenin less than 75% was signi®cantly greater thanwith aberrant staining more than 75% (log-rank test: p=0.0122)

346 T. Saito et al.


tained mutations that altered the potential GSK-3bphosphorylation site (three in codon 32, GAC to TAC;one in codon 37, TCT to TTT; Table 6, Figure 5).Survival data were available for three out of these fourpatients, two of whom suffered early deaths (DOD at 6months and 11 months). However, there was nostatistically signi®cant relationship between b-cateninmutation status and early death (within 1 year,p=0.136). All four cases with b-catenin mutationsshowed aberrant b-catenin expression with reducedexpression of E-cadherin.

Discussion

It has recently been demonstrated in various carcino-mas that E-cadherin and catenins are essential for

intercellular junctions and that their decreased expres-sion corresponds to poor prognosis. The present studyshows for the ®rst time that preserved immuno-reactivity for E-cadherin and a-catenin is signi®cantlycorrelated with a favourable survival rate in synovialsarcoma. Furthermore, along with a low AJC stage

Figure 3. Aberrant nuclear expression of b-catenin wasrecognized in a case of monophasic ®brous synovial sarcomawith b-catenin mutation (case 68, immunohistochemistry)

Figure 4. SSCP analysis in cases with b-catenin mutations insynovial sarcoma. Aberrantly migrating bands were recognized(arrows: aberrantly migrating bands; C: control)

Figure 5. Nucleotide sequence analysis of DNA, corresponding to the aberrantly migrating bands detected by SSCP analysis, showsb-catenin mutations in exon 3 (A, control; B, case 68, codon 32, GAC to TAC; C, case 61, codon 37, TCT to TTT)



and a low necrotic rate, preserved immunoreactivity

for a-catenin was con®rmed as an independent favour-

able prognostic factor using stepwise multivariate

survival analysis with Cox's proportional hazards

model; there was also a signi®cant correlation between

a-catenin expression and low MIB-1 LI. Matsui et al.

[5] have demonstrated that in human gastric cancer,

the frequency of lymph node metastasis and haemato-

genous liver metastasis in E-cadherin-positive/a-cate-

nin-negative tumours was signi®cantly higher than in

E-cadherin-positive/a-catenin-positive tumours. In the

present study, the group with preserved co-expression

of E-cadherin and a-catenin had a much better survival

rate than the group with preserved expression for E-

cadherin only; a-catenin could thus be considered

the most important adhesion molecule in synovial

sarcoma.Since b-catenin may be a multifunctional protein

involved in the wingless/Wnt signal transduction path-

way, we investigated whether its accumulation within

cytoplasm and/or nuclei would affect proliferative

activity. Hugh et al. [11] demonstrated that the wide-

spread nuclear expression of b-catenin was associated

with poor prognosis in primary colorectal cancer; a

signi®cant correlation between widespread aberrant

staining of b-catenin and poor prognosis was noted in

the present study and we were also able to ®nd a close

relationship between the widespread aberrant staining

of b-catenin and MIB-1 LI. Li et al. [9] demonstrated

that APC truncating mutations gave aggressive ®bro-

matosis cells a proliferative advantage through b-

catenin and suggested that b-catenin acted to transac-

tivate transcription. Tetsu et al. [29] recently demon-

strated that abnormal levels of b-catenin may

contribute to neoplastic transformation by causing

accumulation of cyclin D1 in colon carcinoma cells.

These reports support our conclusion that widespread

aberrant staining of b-catenin may contribute to

Table 6. Correlation between b-catenin mutation and prognosis

Case No. Codon Amino acid change MIB-1 LI Prognosis

46 32 (GAC to TAC) Asp to Tyr 5.3 Unknown59 32 (GAC to TAC) Asp to Tyr 11.1 Alive (61 months)

61 37 (TCT to TTT) Ser to Phe 23.6 DOD (11 months)

68 32 (GAC to TAC) Asp to Tyr 26.6 DOD (6 months)

DOD=died of disease.

Table 4. Correlation between the expression of E-cadherin and catenins and clinicopathological parameters

E-cadherin a-catenin b-catenin c-catenin b-catenin aberrant expression

+ x + x + x + x + x

Sex p=0.050* p=0.073 p=0.013* p=0.090 p=0.217

Female (n=37) 14 23 9 28 12 25 7 30 19 18Male (n=35) 6 29 3 32 3 32 2 33 23 12

Glandularity p<0.0001* p<0.0001* p<0.0001* p<0.0001* p=0.0052*

High (n=15) 13 2 9 6 10 5 8 7 4 11Low (n=57) 7 50 3 54 5 52 1 56 38 19

Mitotic rate (per 10 HPF) p=0.011* p=0.120 p=0.051 p=0.055 p=0.619

<15 (n=53) 19 34 11 42 14 39 9 44 30 23>15 (n=19) 1 18 1 18 1 18 0 19 12 7

Nuclear grade p=0.010* p=0.062 p=0.160 p=0.115 p=0.268

I or II (n=58) 20 38 12 46 14 44 9 49 32 26III (n=14) 0 14 0 14 1 13 0 14 10 4

Necrosis p=0.460 p=0.594 p=0.427 p=0.822 p=0.920

<50% (n=58) 15 43 9 49 11 47 7 51 34 24

>50% (n=14) 5 9 3 11 4 10 2 12 8 6

AJC stage p=0.073 p=0.146 p=0.324 p=0.180 p=0.358

I or II (n=37) 13 24 9 28 9 28 6 31 19 18

III or IV (n=22) 3 19 2 20 3 19 1 21 14 8

*Statistically signi®cant.

Table 5. Result of stepwise multivariate survival analysiswith Cox's proportional hazards model

Variable Coef®cient Coef®cient/SE

Hazards

ratio p value

AJC stage 2.1151 4.3781 8.2907 <0.0001

a-catenin x2.746 x2.6299 0.0642 0.0001

Necrosis 1.2687 2.5773 3.5562 0.0139

348 T. Saito et al.


tumour progression through increasing proliferative

activity in synovial sarcoma.The b-catenin mutations found in this study were

similar to those previously reported [15,17,19,25], but

mutations have not so far been linked to prognosis. It

is interesting that two of our three cases with b-catenin

mutations and with available follow-up data died

within 1 year (DOD at 6 and 11 months, respectively).

There was, however, no signi®cant correlation between

b-catenin mutation and early death (Fisher's:

p=0.136). Abnomal levels of b-catenin caused by

other factors, such as APC mutation, may also

in¯uence proliferative activity.The preserved expression of cell±cell adhesion

molecules has previously been correlated with favour-

able prognosis, but relationships with other clinico-

pathological factors have not been clearly shown. In

this study, the preserved expression of E-cadherin was

signi®cantly correlated with sex, low mitotic rate (<15

per 10 HPF), and low nuclear grade (grades I and II),

most of which are recognized prognostic factors in

synovial sarcoma.Our immunohistochemical analysis revealed aber-

rant staining of b-catenin protein to a variable degree

in more than half of our cases of synovial sarcoma (42/

72), regardless of the type of E-cadherin expression. If

we take this into consideration, the group with b-

catenin accumulation due most likely to the inactiva-

tion of b-catenin itself, or APC, becomes much more

smaller (7/72) than the group in which abnormal levels

of b-catenin were mainly considered to be due to the

failure of E-cadherin to localize at the cellular

membrane (35/72). However, all four cases with b-

catenin mutation in this study belonged to the latter

group. Although our DNA analysis was very limited,

our results suggest that b-catenin mutation is infre-

quent in synovial sarcoma; an unknown mechanism of

b-catenin regulation may be involved.In conclusion, the present study suggests that E-

cadherin and a-catenin play important roles in synovial

sarcoma and their expression could assist in the

prediction of prognosis. Furthermore, abnormal levels

of b-catenin in tumour cells, regardless of the presence

of mutation, are considered to lead to oncogenic b-

catenin activation, contributing to tumour progression

in synovial sarcoma.

Acknowledgements

This work was supported in part by a Grant-in-Aid for Cancer

Research from the Fukuoka Cancer Society, Fukuoka; a Grant-

in-Aid for General Scienti®c Research from the Ministry of

Education, Science, Sport and Culture (09470052), Tokyo; and a

Grant-in-Aid for Scienti®c Research from the Ministry of

Education, Science, Sports and Culture (10307034 and

09557124), Japan. We are grateful to Miss K. Nishiyama for

her excellent technical assistance. We thank Miss Katherine

Miller (Royal English Language Centre, Fukuoka, Japan) for

revising the English used in this article.

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Prognostic value of the preserved expression of the E-cadherin and catenin families of adhesion molecules and of β-catenin mutations in synovial sarcoma