Analysis of APC, α-, β-catenins, and N-cadherin protein expression in aggressive fibromatosis (desmoid tumor)

ARTICLE IN PRESS

0344-0338/$ - se

doi:10.1016/j.pr

$This work w

National Comm�CorrespondiE-mail addre

Pathology – Research and Practice 205 (2009) 311–324

www.elsevier.de/prp

ORIGINAL ARTICLE

Analysis of APC, a-, b-catenins, and N-cadherin protein

expression in aggressive fibromatosis (desmoid tumor)$

Tomasz Ferenca,�, Jan Wojciech Wronskib, Janusz Kopczynskic, Andrzej Kuligd,Małgorzata Sidorc, Liliana Stalinskaa, Adam Dzikie, Jacek Sygutc

aDepartment of Biology and Genetics, Medical University, Pl. Hallera 1, 90-647, Lodz, PolandbDepartment of General Surgery, Provincial Podkarpacki Hospital, Krosno, PolandcDepartment of Neoplasm Pathology, Swietokrzyski Centre of Oncology, Kielce, PolanddDepartment of Clinical Pathomorphology, Institute Polish Mother’s Health Centre, Lodz, PolandeDepartment of General and Colorectal Surgery, Medical University, Lodz, Poland

Received 3 July 2008; received in revised form 4 November 2008; accepted 6 November 2008

Abstract

The aims of this study were to analyze the cadherin/catenin adhesion complex in cells from abdominal and extra-abdominal aggressive fibromatosis tumors, and to estimate the correlation between the expression of the testedproteins and the clinical data of the desmoid patients.

Immunohistochemistry was used to examine the expression of the cadherin/catenin adhesion complex: APC protein,a-, b-catenin, and N-cadherin in archival material derived from 15 cases of extra-abdominal desmoid tumor (E-AD)and 20 cases of abdominal (AD) desmoid tumor. The tested proteins demonstrated cytoplasmic (c) staining.Furthermore, nuclear (n) or cytoplasmic and nuclear (c+n) staining was observed for b-catenin. The mean values ofthe percentage of positive cells for the tested proteins between E-AD vs. AD did not demonstrate any statisticallysignificant difference except for a-catenin. In the E-AD group, in both cases of recurrent tumors, no a-cateninexpression was observed but the expression of this protein was detected in primary tumors. In the groups investigated,no statistically significant correlation was found between a-catenin, b-catenin (c), (n) and (c+n) expression, and tumorsize (p40.1).

The results regarding b-catenin expression obtained in our study confirm the previous findings that nuclearaccumulation of this protein plays a crucial role in the pathogenesis of aggressive fibromatosis.r 2008 Elsevier GmbH. All rights reserved.

Keywords: Aggressive fibromatosis (desmoid); APC; a-, b-catenin, N-cadherin; Immunohistochemistry

e front matter r 2008 Elsevier GmbH. All rights reserved.

p.2008.11.002

as supported by the Grant no. 3P05A 033 24 from the

ittee for Scientific Research, Poland.

ng author. Tel./fax: 0048 042 6330594.

ss: [email protected] (T. Ferenc).

Introduction

Desmoid-type fibromatoses are clonal fibroblasticproliferations that arise in the deep soft tissues, andare characterized by infiltrative growth and a tendencytowards local recurrence but an inability to metastasize[20]. Due to location, aggressive fibromatosis can bedivided into several groups: extra-abdominal (E-AD),

www.elsevier.de/prp

dx.doi.org/10.1016/j.prp.2008.11.002

mailto:[email protected]

ARTICLE IN PRESST. Ferenc et al. / Pathology – Research and Practice 205 (2009) 311–324312

abdominal (AD), and intra-abdominal (I-AD) [54].Being dependent on its location in various parts of thebody, it demonstrates a different biological behavior[23,44,51]. This rare benign tumor occurs in sporadicforms [3,4,50], and is also associated with familialadenomatous polyposis (FAP) [7,15,16,28].

The adenomatous polyposis coli (APC) protein(312 kDa) is multifunctional (e.g., cell migration andapoptosis), and appears to be linked to the breakdownof b-catenin. The APC protein plays an essential role inclearing unnecessary b-catenin from the cytoplasm, andb-catenin acquires oncogenic activity when it is mutatedor upregulated as a result of inactivation of APC[18,24,42]. APC gene mutations in somatic cells havebeen reported to play an important role in thedevelopment of aggressive fibromatosis sporadic tumors[3,4,18]. Germline mutations of the APC gene have beenfound mainly in exon 15 between codons 1035 and 1581in patients with FAP-associated aggressive fibromatosis[15,16,28].

The a-catenin (102 kDa) is associated with thecarboxy-terminal region of the cadherin cytoplasmicdomain via b-catenin to form a functional cadherin–catenin cell adhesion complex [24,35]. Three a-cateninsubtypes have been described: aE-catenin, aN-cadherin,and aT-catenin [6]. Thus, loss of a-catenin expressionmay lead to impaired formation of adherens junctionsdespite normal E-cadherin and b-catenin expression[56]. The reduction of a-catenin expression has beencorrelated with tumor dedifferentiation, infiltrativegrowth, and lymph node metastasis in various humancarcinomas [6,10,13,46,47,56].

The b-catenin (92 kDa) is a multifunctional proteininvolved in the wingless/Wnt signal transduction path-way [27,34]. It exists in three different subcellular forms[33,55]. It has a nuclear function and binds transcriptionfactors with the T-cell factor/lymphoid-enhancingfactor (Tcf-Lef) family and a cell membrane function.b-catenin can complex with other molecules andcontains binding sites for a-catenin, APC, andE-cadherin. Cytoplasmic b-catenin exits in a solubleand monomeric state and can be degraded or translo-cated to the nucleus [27,33]. b-catenin is degraded by theubiquitin–proteosome system after complexing withAPC protein, glycogen synthase kinase (GSK3b), andaxin/conduction [24,27,33]. Abnormal nuclear locationof b-catenin and its association with Tcl-Lef have beenproposed as an important oncogenic step in variousbenign and malignant fibroblastic and myofibroblastictumors [3,8,12,32,39,49,52].

The N-cadherin (135 kDa) is a transmembraneglycoprotein and a member of the Ca2+-dependentcell–cell adhesion molecule family. The cytoplasmicdomain of N-cadherin is anchored to the intercellularactin cytoskeleton by interacting with the a-, b-, andg-catenin complex [14]. N-cadherin is expressed mainly

in the nervous system, mesenchymal tissues, muscle, lensepithelial cells, and fibroblasts [22,25]. In embryonicdevelopment, N-cadherin plays an essential role inmyoblast recognition and adhesion during skeletalmyogenesis [29]. Recent studies have demonstrated thataberrant N-cadherin expression in tumor tissues isassociated with tumor progression, such as epithelial–mesenchymal transition (EMT), motility, and metastasis[2,9,21,40,41].

The purpose of this study was to analyze the cadherin/catenin adhesion complex (APC protein, a-, b-catenin,N-cadherin) with immunohistochemical methods in cellsfrom AD and E-AD aggressive fibromatosis tumors.Furthermore, we estimated the results regarding thecorrelation between the expression of the tested proteinsand the clinical data of desmoid patients.

Materials and methods

Tumor specimens

Formalin-fixed, paraffin-embedded archival tissuesobtained from 35 cases of sporadic aggressive fibroma-tosis (desmoid) were studied: 20 AD (20 females; meanage 30.278.3 years) and 15 E-AD cases (including2 recurrent tumors) (10 females and 5 males; mean age38.5716.0 years). The archival specimens were fromthe Departments of Pathomorphology in Poland.* Allthe sections were examined independently by twopathologists (AK, JS) using a conference microscope,and were histopathologically classified as recommendedby the World Health Organization Classification ofTumors (2002) [20] and Weiss, Goldblum [54]. Thestudy was carried out with the approval of the localethics committee.

Immunohistochemical staining

Representative paraffin blocks containing tumormaterial from each case were sectioned at 4 mm, affixedto silanized slides, and dried overnight at 60 1C. Thetissue sections were deparaffinized and rehydrated. Anantigen retrieval was performed by pretreatment inTarget Retrieval Solution Citrate (pH 6.0) (DAKOCytomation, DK) in a water bath (95–99 1C, 40min).The sections were subsequently rinsed with 3% hydro-gen peroxide for 10min at room temperature and werethen incubated with primary mouse monoclonal anti-bodies for 1 h at room temperature. The antibodies(including polyclonal) used for immunohistochemicalstaining and their dilutions are demonstrated in Table 1.The protein expression was detected using LSABs+SYSTEM-HR reagent (DAKO, Cytomation, DK).3,30-diaminobenzidine (Liquid DAB+Chromogen)

ARTICLE IN PRESS

Table 1. Summary of the antibodies used in the immunohistochemical analysis of desmoid tumors.

Antibody Clone Dilution Company

APC EMM43 1:20 Novocastra New Castle upon Tyne, UK

Alpha-catenin 25B1 1:50 Novocastra New Castle upon Tyne, UK

Beta-catenin 17C2 1:50 Novocastra New Castle upon Tyne, UK

CD34 QBEend/10 1:50 Novocastra New Castle upon Tyne, UK

N-cadherin Polyclonal 1:200 Takara, Bio INC, Otsu, Shiga, Japan

Fig. 1. Immunoreactivity of a-catenin (A), b-catenin (B), N-cadherin (C), APC (D) in desmoid tumor. Representative images of

positive expression (identified by brown staining) are shown. Nuclei were counterstained with Mayer’s hematoxylin. Original

magnification: � 200 (A–C), � 100 (D).

T. Ferenc et al. / Pathology – Research and Practice 205 (2009) 311–324 313

(DAKO) was used as chromogen to yield brownreaction product (Fig. 1A–D). The sections werecounterstained with Mayers hematoxylin, dehydrated,and mounted. The paraffin-embedded tonsil sectionswere used as a positive control for APC, a-catenin,b-catenin, and N-cadherin. Omission of the primaryantibody and substitution by non-specific immunoglo-bulins were used as negative controls. The immunohis-

tochemical reaction for CD34 [37,39] (clone QBEnd/10,Novocastra, UK) was performed for each case.

Immunohistochemical analysis

A semiquantitative approach was used for scoringthe immunostaining [53]. Evaluation and scoring of

ARTICLE IN PRESS

Table 2. Summary of the clinical data of 15 patients with

extra-abdominal desmoid tumors (E-AD).

Case no. Age/Sex Anatomical

location

Largest tumor

dimension (cm)

1. 15/M Popliteal fossa 7.0

2. 25/M Thoracic wall 3.0

3. 29/F Spinal region 4.0

4. 30/F Thoracic wall 3.0

5.P 27/M Thoracic wall 4.5

5.R 30/M Thoracic wall 4.5

6. 30/F Coxa 5.0

7. 35/M Neck 6.0

8.P 43/F Arm 8.0

8.R 46/F Arm 6.0

9. 62/M Thoracic wall 2.0

10. 28/F Arm NA

11. 68/F Thoracic wall 2.5

12. 33/F Buttock 2.0

13. 65/F Back 6.0

F ¼ female; M ¼ male; P ¼ primary lesion; R ¼ recurrent lesion;

NA ¼ not available.

Table 3. Summary of the clinical data of 20 patients with

abdominal desmoid tumors (AD).

Case no. Age/Sex Anatomical

location

Largest tumor

dimension (cm)

T. Ferenc et al. / Pathology – Research and Practice 205 (2009) 311–324314

immunostaining were undertaken independently by twopathologists (JS and JK) whenever necessary; a con-sensus was reached using a double-headed microscope.According to staining intensity, the cases were classifiedas (�) when there was no expression, as (+) whenstaining was very weak, as (++) when it was strong butstill weaker than the positive control, and as (+++)when the staining intensity matched the control [46].The immunostaining was also classified according to thelocation as cytoplasmic and/or nuclear. For all anti-bodies, the cases scored X10% of stained cells wereconsidered positive. The percentage of positive cells wascounted in each examined case in 10 high-power fields(HPF) with � 40 magnification of the objective lens.

Statistical analysis

The basic statistical analysis (arithmetic mean, med-ian, range, quantiles, standard deviation, standarderror) was performed. Pearson Correlation Coefficient

(RP) was calculated to analyze the percentage ofpositively stained cells in each group investigated andto estimate the dependence between b-catenin expres-sion and tumor size or patient age. Student’s t-test wasused to compare the mean values of the percentage ofpositive cells between the groups examined. po0.05 wasregarded as statistically significant. The statisticalanalysis was performed using SAS software.

1. 25/F Abdominal wall 4.0

2. 38/F Musculus rectus 5.0


4. 22/F Abdominal wall NA






10. 26/F Abdominal wall NA











F ¼ female; NA ¼ not available.

Results

The clinical data of the examined cases of desmoids ofE-AD and AD location are given in Tables 2 and 3. InE-AD, the cases 5P and 8P refer to the patients withprimary tumors, and the cases 5R and 8R to the patientswith recurrent tumors.

Immunohistochemical analysis

Alpha-catenin-positive cytoplasmic (c) staining ofa-catenin was detected in 10 of 15 (68%) E-AD casesand in 13 of 20 (65%) AD cases. The mean values of thepercentage of a-catenin (c)-positive cells in the groupsstudied are presented in Table 4. The comparison of themean values of the percentage of a-catenin (c)-positivecells between E-AD vs. AD demonstrated a statisticallysignificant difference (p ¼ 0.0165). The patterns ofa-catenin expression for particular cases in the studiedgroups are presented in Tables 5 and 6. Five E-AD cases(5R, 6, 8R, 11 and 13) and seven AD cases (1, 2, 4, 12,13, 17 and 18) lacked a-catenin cytoplasmic staining.In all positive cases of the studied groups (except case 2in E-AD) was cytoplasmic a-catenin staining intensityclassified as (+), (Tables 5 and 6).

There were no cases with nuclear staining of a-cateninin the groups investigated. In E-AD and AD, nostatistically significant correlation was noted betweenthe expression of a-catenin (c) and b-catenin (c), (n),(c+n) (p40.1); a-catenin (c) and N-cadherin (c)

ARTICLE IN PRESS

Table 4. The mean percentage of APC, a-catenin, b-catenin, and N-cadherin in the study groups.

Tumor type APC a-catenin b-catenin N-cadherin

Extra-abdominal (n ¼ 15)a 0.00 (0.0) 61.5 (719.4) c 51.3 (727.7) c 55.0 (733.2) c

24.7 (734.6) n

24.0 (735.0) c+n

Abdominal (n ¼ 20) 27.5 (712.6) c 42.3 (715.9) c 35.5 (723.3) c 42.5 (732.0) c

32.5 (724.9) n

26.0 (727.8) c+n

n ¼ nucleus; c ¼ cytoplasm; c+n ¼ cytoplasm and nucleus.aIncluding two recurrent lesions.

Table 5. Immunohistochemical expression of APC, a-catenin, b-catenin, and N-cadherin in 20 patients with abdominal desmoid

tumors (AD).

APC a-catenin b-catenin N-cadherin

Case Cyt % of cells Cyt % of cells Cyt % of cells Nuc % of cells Cyt % of cells

1. 0 0 0 0 + 20 + 20 0 0

2. 0 0 0 0 0 0 + 30 + 70

3. + 30 + 30 ++ 40 ++ 40 0 0

4. 0 0 0 0 + 40 + 40 0 0

5. 0 0 + 30 0 0 + 30 0 0

6. 0 0 + 50 + 40 0 0 0 0

7. 0 0 + 60 0 0 + 50 0 0

8. 0 0 + 30 + 60 + 60 0 0

9. 0 0 + 70 + 60 + 60 0 0

10. 0 0 + 40 + 20 0 0 ++ 70

11. 0 0 + 40 ++ 60 0 0 0 0

12. 0 0 0 0 + 20 0 0 0 0

13. 0 0 0 0 + 70 + 70 0 0

14. + 30 + 60 ++ 80 ++ 80 0 0

15. 0 0 + 20 + 20 + 20 0 0

16. + 10 + 50 ++ 40 ++ 40 0 0

17. 0 0 0 0 + 20 + 20 0 0

18. 0 0 0 0 + 40 + 40 0 0

19. + 40 + 50 + 30 0 0 + 20

20. 0 0 + 20 + 50 + 50 + 10

Cyt ¼ cytoplasm; Nuc ¼ nucleus.


(p40.1), and between a-catenin (c) and APC in ADlocation (p40.1) (Tables 7 and 8).

APC-positive cytoplasmic staining of APC proteinwas found in 4 of 20 (20%) AD cases. No positivestaining of APC protein was detected in E-AD cases.The mean values of the percentage of APC-positive cellsin the investigated groups are presented in Table 4. Thepatterns of APC expression for particular cases in theinvestigated groups are demonstrated in Table 5. In ADdesmoid fibromatosis, APC-positive cases expressed thestaining intensity classified as (+).

Tables 7 and 8 show calculated correlation coeffici-ents between all the analyzed proteins in the studiedgroups. In AD, no statistically significant correlationwas observed between APC expression and a-catenin

(c) (p40.1) and between APC and b-catenin (c), (n) and(n+c) (p40.1) (Table 7).

b-catenin-positive cytoplasmic and/or nuclear stain-ing of b-catenin was observed in 15 of 15 (100%) E-ADcases and in 20 of 20 (100%) AD cases. The mean valuesof the percentage of b-catenin cytoplasmic and/ornuclear staining in the study groups are presented inTable 4. In addition, we analyzed the expression ofb-catenin in five I-AD cases (mean age 28.2 7 6.9 years).In the I-AD group, the mean values of the positive cellsfor b-catenin (c), (n), and (c+n) were 50.0 (733.9), 42.0(735.6), and 38.8 (737.7), respectively. We made acomparison of the mean values of the percentage ofpositive cells for b-catenin (c), (n), and (c+n) betweenthe groups: E-AD vs. AD did not show any statistically

ARTICLE IN PRESS

Table 6. Immunohistochemical expression of a-catenin, b-catenin, and N-cadherin in 15 patients with extra-abdominal desmoid

tumors (E-AD).

a-catenin b-catenin N-cadherin

Case Cyt % of cells Cyt % of cells Nuc % of cells Cyt % of cells

1. + 80 + 10 + 10 0 0

2. ++ 90 ++ 80 ++ 80 0 0

3. + 60 ++ 80 ++ 80 0 0

4. + 40 +++ 80 +++ 80 + 10

5.P + 60 ++ 40 ++ 40 0 0

5.R 0 0 + 20 0 0 0 0

6. 0 0 + 40 0 0 ++ 60

7. + 50 + 80 0 0 0 0

8.P + 30 ++ 60 ++ 60 0 0

8.R 0 0 + 30 0 0 0 0

9. + 80 ++ 60 0 0 0 0

10. + 50 ++ 70 ++ 70 ++ 60

11. 0 0 + 40 0 0 0 0

12. + 75 + 80 0 0 +++ 90

13. 0 0 0 0 + 10 0 0

P ¼ primary lesion; R ¼ recurrent lesion; Cyt ¼ cytoplasm; Nuc ¼ nucleus.

Table 7. The results of the comparison of APC, a-, b-catenins and N-cadherin expression in abdominal location (AD).

APC (c) a-catenin (c) b-catenin (c) b-catenin (n) b-catenin (c+n) N-cadherin (c)

APC (c) �0.0526 �0.0299 �0.3244 �0.3244 –

0.9474 0.9701 0.6756 0.6756 –

a-catenin (c) �0.0526* 0.2059 0.2136 0.1061 0.3394

0.9474** 0.4998 0.4836 0.7300 0.7795

b-catenin (c) �0.0299 0.2059 0.5112 0.7923 �0.8753

0.9701 0.4998 0.0212 o0.0001 0.1247

b-catenin (n) �0.3244 0.2136 0.5112 0.8672 �0.3400

0.6756 0.4836 0.0212 o0.0001 0.6600

b-catenin (c+n) �0.3244 0.1061 0.7923 0.8672 �0.6768

0.6756 0.7300 o0.0001 o0.0001 0.3232

N-cadherin (c) – 0.3394 �0.8753 �0.3400 �0.6768

– 0.7795 0.1247 0.6600 0.3232

*Pearson Correlation ¼ RP; **p ¼ value; c ¼ cytoplasm; n ¼ nucleus; c+n ¼ cytoplasm and nucleus.


significant differences (p40.1) except for the compar-ison for b-catenin (c), where the difference was on theborder of statistical significance (p ¼ 0.0756).

The observed patterns of b-catenin staining forparticular cases of the investigated groups are presentedin Tables 5 and 6. Positive cases for b-catenin (c), (n),and (c+n) in the group studied demonstrated differentintensity of staining (+) or (++). Only in case 4, in

extra AD location, staining intensity was (+++) forb-catenin (c+n). Nuclear staining of b-catenin (n) wasdetected for case 13 in E-AD and for cases 2, 5, and 7 inAD. Cytoplasmic staining (c) was noted in 8 E-AD cases(5R, 6, 7, 8P, 8R, 9, 11 and 12) and in 5 AD cases (6, 10,11, 12 and 19). In the remaining cases of the studiedgroups, both cytoplasmic and nuclear staining ofb-catenin was observed.

ARTICLE IN PRESS

Table 8. The results of the comparison of a-, b-catenins and N-cadherin expression in extra-abdominal location (E-AD).

a-catenin (c) b-catenin (c) b-catenin (n) b-catenin (c+n) N-cadherin (c)

a-catenin (c) �0.2119 �0.0363 �0.0363 0.9265

0.5567 0.9207 0.9207 0.2457

b-catenin (c) �0.2119* 0.5064 0.5384 �0.1327

0.5567** 0.0541 0.0384 0.8673

b-catenin (n) �0.0363 0.5064 0.9973 �0.7510

0.9207 0.0541 o0.0001 0.2490

b-catenin (c+n) �0.0363 0.5384 0.9973 �0.7510

0.9207 0.0384 o0.0001 0.2490

N-cadherin (c) 0.9265 �0.1327 �0.7510 �0.7510

0.2457 0.8673 0.2490 0.2490

*Pearson Correlation ¼ RP; **p ¼ value; c ¼ cytoplasm; n ¼ nucleus; c+n ¼ cytoplasm and nucleus.


Positive correlation coefficients were noted forb-catenin, and they were statistically significant betweenthe comparisons: b-catenin (c) and (n) (p ¼ 0.0541);b-catenin (c) and (c+n) (p ¼ 0.0384); b-catenin (n) and(n+c) (p ¼ 0.0001) in E-AD and b-catenin (c) and (n)(p ¼ 0.0212); b-catenin (c) and (n+c) (po0.0001);b-catenin (n) and (c+n) (p ¼ 0.0001) in AD (Tables 7and 8).

N-cadherin-positive cytoplasmic (c) staining ofN-cadherin was found in 4 of 15 (27%) E-AD casesand in 4 of 20 (20%) AD cases. The mean values of thepercentage of positive cells for N-cadherin (c) in thestudied groups are presented in Table 4. The compar-ison of the mean value of the percentage of positivecases for N-cadherin (c) between E-AD vs. AD did notdemonstrate a statistically significant difference(p ¼ 0.6071). In the studied groups, the positive casesfor N-cadherin (c) showed different staining intensityexpressed as (+), (++) or (+++) (Tables 5 and 6).

In E-AD and AD, no statistically significant correla-tion was noted between the expression of N-cadherin(c) and a-catenin (c), (p40.1); N-cadherin (c) andb-catenin (c), (n), (n+c) (p40.1), and in AD locationbetween N-cadherin (c) and APC (p40.1).

b-catenin expression with tumor size and age

Considering that b-catenin expression was observed inall cases in the studied groups, an analysis of thecorrelation between b-catenin (c), (n) and (c+n) andE-AD and AD tumor size was performed. In E-AD,there was no statistically significant correlation between

b-catenin expression (c), (n) and (c+n) and tumor size,respectively (Rp ¼ �0.4518, p ¼ 0.1048), (Rp ¼ �0.3038,p ¼ 0.2910) and (Rp ¼ �0.3180, p ¼ 0.2677) (Fig. 2).Also in AD, there was no statistically significantcorrelation between b-catenin expression (c), (n) and(c,+n) and tumor size, respectively (Rp ¼ �0.0841,p ¼ 0.7480), (Rp ¼ 0.1349, p ¼ 0.6057) and (Rp ¼ �0.0812,p ¼ 0.7565) (Fig. 3).

Fig. 4A–C illustrate the results of the correlationbetween the expression of b-catenin (c), (n) and (c+n)and patient age in E-AD, AD, and I-AD location(total). A statistically significant correlation was ob-served for b-catenin (n) and (c+n), respectively(Rp ¼ �0.3377, p ¼ 0.0330) and (Rp ¼ �0.3189,p ¼ 0.0449) (Fig. 4B, C). No statistical significance wasdetected for b-catenin (c) (Rp ¼ �0.0523, p ¼ 0.7485)(Fig. 4A).

Discussion

The analysis of expression and interactions ofcadherins, including N-cadherin, with a-catenin andb-catenin forming a functional cadherin–catenin unit(ECCU), and APC protein, has already been subject ofnumerous studies of human neoplasms derived fromdifferent tissues [10,13,19,26,46,48].

So far, there has been a lack of studies analyzing theexpression of a complex of adhesive proteins inaggressive fibromatosis tumor cells using immunohisto-chemical methods. In this study, we analyzed theexpression of APC protein, a-, b-catenin, and

ARTICLE IN PRESS

Fig. 2. Comparison of the expression of b-catenin (c), (n) and (c+n) and tumor size, in extra-abdominal location (E-AD).

*including two recurrent cases.


N-cadherin in desmoid cells of 15 cases in E-AD and of20 cases in AD location. Furthermore, the results of thecorrelation between the expression of the tested proteinsand the clinical data of desmoid patients were estimated.The observed patterns of expression as well as the resultsof comparisons of the percentage of positive cells for theinvestigated proteins in the E-AD and AD groups andcomparisons between E-AD vs. AD were different. Thismight result from certain differences in the biology ofthese two forms of the neoplasm. Individual variabilityin the dynamics of development and progression ofaggressive fibromatosis has also been indicated.

Although AD fibromatosis is indistinguishable fromE-AD fibromatosis as regards macroscopic and micro-scopic structure and infiltrating growth, some differ-ences in clinical pathology should be indicated. ADfibromatosis is most frequently observed in youngwomen, particularly in those who have already had

one or more children. The peak incidence occurs at theage of 20–30 years [54]. The rectus and the internaloblique muscles of the abdomen are the most frequentlyaffected sites. Tumors of abdominal location are usuallyof smaller size and of lesser destructive effect on thesurrounding tissues than tumors of E-AD location. Inturn, E-AD fibromatosis can be located at differentanatomical sites. E-AD tumors most frequently occurbetween puberty and 40 years of age. Peak prevalence isat the age of 25–35 years [54].

APC protein

APC gene mutations have been confirmed both insporadic and in FAP-associated desmoid tumors[3,7,16,18,28,38]. A major function of APC protein isto regulate the b-catenin protein level. In the study of

ARTICLE IN PRESS

Fig. 3. Comparison of the expression of b-catenin (c), (n) and (c+n) and tumor size in abdominal location (AD).


Li et al. [32], transient transfection of the full-lengthAPC gene caused decreased proliferation and decreasedb-catenin protein level in cell cultures from aggressivefibromatosis containing APC mutation. The studies ofAPC protein with the use of immunohistochemicalmethods in cells from normal tissues of different organsdemonstrated certain organ specificity as regards theexpression of this protein and lack of APC expressionin some tissues [36]. So far, in the available literature,only the study of Alman et al. [3] has dealt with APCprotein expression in aggressive fibromatosis tumorswith the use of immunohistochemical methods. Almanet al. analyzed APC protein expression using amino-and carboxyl-terminal antibodies to APC (N-APC andC-APC) in six cases of aggressive fibromatosis ofthe extremities. In two cases, expression was noted onlyfor the antibody detecting the amino-terminal region ofAPC, and in two cases, staining was found for both thecarboxyl-terminal and the amino-terminal region of

APC. In the two remaining cases, it was not possible tointerpret the results [3]. In our study, lack of theexpression of APC protein was recorded in 15 of 15(100%) E-AD cases and in 16 of 20 (80%) AD cases.The question arises whether the low number of caseswith positive APC staining observed in our immunohis-tochemical analysis resulted from mutational truncationof functional APC region or from low expression ofAPC protein, resulting from the embryonic derivation ofcells of which the desmoid tumor is built. Midgley et al.[36] evaluated the expression of APC protein (amino-and carboxyl-terminal) in skeletal muscle cells as (7).

a-catenin

Decreased expression of a-catenin has been reportedin various carcinomas, e.g. of the bladder [13], breast[19], colorectum [47], prostate [26], synovial sarcoma

ARTICLE IN PRESS

Fig. 4. Comparison of the expression of b-catenin (c), (n) and (c+n) in all the investigated groups and patient age. *Five cases of

I-AD included.


[48], and thyroid [10,46]. In some of the studies, acorrelation was suggested between a-catenin expressionand dedifferentiation, infiltrative growth, and metasta-sis. Due to the lack of other reports concerning theanalysis of a-catenin expression in aggressive fibroma-tosis tumors, it is impossible to make any comparisons.In this study, the lack of a-catenin (c) expression wasobserved in 5 of 15 (33%) E-AD cases and in 7 of 20(35%) AD cases. Bohm et al. analyzed membrane (m)expression of a-catenin in differentiated thyroid carci-noma. They observed abnormal membrane staining fora-catenin in 40% of papillary carcinomas and in 52% offollicular neoplasms. They reported that decreasedexpression of a-catenin in differentiated thyroid carci-nomas was associated with recurrence of these tumors[10]. In our study, a-catenin (c) was not expressed in twocases of recurrent desmoid tumor of E-AD location(cases 5R and 8R) and in two cases of primary tumors

(cases 6 and 11). However, in primary tumors (cases 5Pand 8P), the percentage of cells with a-catenin (c)staining was 60% and 30%, respectively. In case 6, therecurrence of the tumor was detected in the iliac region,and in case 11, the tumor developed after mastectomy.

Lack of a-catenin (c) expression was detected in sixE-AD tumors (cases 1, 2, 12, 13, 17, 18). The duration ofthe follow-up period was 2–6 years after tumor surgery,and within this period, no recurrences were observedexcept for case 17. Kallakury et al. [26] revealed lack ofmembrane staining for a-catenin in 17% of cases withprostatic adenoma, and decreased expression ofa-catenin correlated with high tumor grade and aneu-ploidy. Clairotte et al. [13] noted a highly significantcorrelation between the decreased membrane (m)expression of a-catenin and increased TNM stage inbladder cancer. In our study, summarizing the caseswith extra-abdominal and abdominal location of

ARTICLE IN PRESST. Ferenc et al. / Pathology – Research and Practice 205 (2009) 311–324 321

desmoid tumor, a negative correlation was observedbetween tumor size and expression of a-catenin (c).However, the results of this correlation were notstatistically significant (Rp ¼ �0.0521, p ¼ 0.8274).

b-catenin

In normal fibroblast and endothelial cells, b-cateninstaining is limited to cytoplasm and/or cell membrane[45]. Abnormal nuclear localization of b-catenin and itsassociation with Tcl-Lef were proposed as an importantoncogenic step in various benign and malignantfibroblastic and myofibroblastic tumors [3,8,11,12,43,50,52].In our study, only nuclear b-catenin staining (n) wasobserved in one E-AD and in three AD cases (11% of allexamined cases). However, b-catenin staining (c+n) wasnoted in seven E-AD cases and in 12 AD cases, i.e., in55% of the examined cases. In the study of Bhattachar-ya et al., all the 21 examples of deep fibromatoses(100%) showed nuclear staining for b-catenin. All deepfibromatoses cases also demonstrated focal to diffusecytoplasmic staining [8]. Amary et al. [5] noted that 66(100%) cases of desmoid-type fibromatosis analyzedshowed unequivocal nuclear positivity for b-catenin in asignificant (425%) number of the tumor cells, and over50% of the cases showed cytoplasmic reactivity. Saitoet al. observed nuclear staining in 19 of 38 (50%) casesof sporadic desmoid tumor of E-AD and AD location.They did not detect any statistically significant differ-ence between nuclear expression for b-catenin and thetumor location (abdominal wall or extra-abdominal)[49]. As in our study, the comparison of the mean valuesof the percentage of cells positive for b-catenin (c), (n)and (c+n) between the examined groups did not displayany statistically significant difference (p40.1). In thestudy of Montgomery et al. [39], nuclear b-catenin wasdetected in 50–95% of nuclei in 9 of 10 fibromatoses,including one case associated with FAP. Carlsonand Fletcher detected nuclear immunopositivity forb-catenin in 80% of cases of sporadic desmoidfibromatosis (24/30) and in 67% of tumors in patientswith FAP (8/12). No significant differences in immunor-eactivity were noted between sporadic and familialdesmoid fibromatosis [11]. Ng et al. [43] revealed nuclearstaining for b-catenin in 12 of 17 (71%) desmoid cases.In our study, nuclear expression for b-catenin (n) and(c+n) was observed in 46% of E-AD and in 75% of ADcases. We did not observe any statistically significantcorrelation between the expression of b-catenin (n) and(c+n) and tumor size in E-AD and AD tumor location(p40.1). On the other hand, Gebert et al. [17] found outthat overexpression (420% positive nuclei) of b-cateninwas associated with a decreased event-free survival indeep aggressive fibromatosis.

Spontaneous regression of these tumors has beenobserved in sporadic cases, particularly in menarchaland menopausal patients [54]. It is interesting that in thisstudy, a statistically significant correlation for the wholeinvestigated group (including five I-AD patients) be-tween the expression of b-catenin (n) and (c+n) andpatient age was observed (po0.05). The mean value ofthe percentage of b-catenin-positive cells decreased withage.

N-cadherin

N-cadherin plays important roles in various biologicalbehaviors, including morphogenesis and angiogenesisduring development [41] and, as emphasized by numer-ous authors, in epithelial–mesenchymal transition(EMT) during tumor progression [1,14,40,41]. Due tothe lack of reports concerning N-cadherin analysis inaggressive fibromatosis tumors using immunohisto-chemistry, it is impossible to make any comparisons.However, in relation to other neoplasms, Nakashimaet al. detected membrane expression for N-cadherin in46 of 150 (30.7%) non-small-cell lung cancer cases.These authors observed that the frequency of hypervas-cular tumors was significantly higher for N-cadherin-positive carcinomas than for N-cadherin-negativecarcinomas (p ¼ 0.0373) [41]. In the studies of Nakajimaet al. [40], investigating pancreatic carcinoma, cytoplas-mic N-cadherin expression was observed in 13 of 30(43.3%) primary tumors and in 8 of 15 (53.3%)metastatic tumors. Kovacs et al. [30] observed cytoplas-mic N-cadherin expression in 30% of invasive breastcarcinomas and in 4 of 10 ductal carcinamas in situ. Inour study, positive cytoplasmic N-cadherin staining wasfound in 4 of 15 (27%) E-AD cases and in 4 of 20 (20%)AD cases. In the study of Lascombe et al., N-cadherin(m) expression was absent in normal urothelium,appeared in stage pT1, and increased in pT2–pT3urothelial tumors. Progression-free survival and multi-variate analyses revealed that N-cadherin expressionwas an independent prognostic marker for pT1 urothe-lial tumor progression [31].

The results of b-catenin expression obtained in ourstudy confirm the previous findings that nuclearaccumulation of this protein plays a crucial role in thepathogenesis of aggressive fibromatosis. This conceptionmight also be supported by the revealed negativecorrelation between the expression of b-catenin (n) and(c+n) and patient age.

Acknowledgements

This work was supported by the Grant no. 3P05A 03324 from the National Committee for Scientific Research,


Poland. *The authors of this research thank the belowmentioned Heads of Chairs and Departments forproviding paraffin blocks and available data for therealization of these studies: The Chair of Pathomor-phology, Collegium Medicum of Jagiellonian Universityin Krakow; The Department of Neoplasms Pathology,Center of Oncology, M. Skłodowska-Curie Institute,Krakow; The Department of Neoplasms Pathology,Center of Oncology, M. Skłodowska-Curie Institute,Gliwice; The Chair and Department of PathologicalAnatomy, Medical University in Białystok; The Chairand Department of Clinical Pathomorphology,K. Marcinkowski Medical University in Poznan; TheChair and Department of Pathological Anatomy,Medical University in Gdansk; The Chair and Depart-ment of Pathological Anatomy, Silesian MedicalUniversity in Katowice; Department of Pathomorpho-logy, Provincial Hospital in Rzeszow.

References

[1] M.L. Ackland, D.F. Newgreen, M. Fridman,

M.C. Waltham, A. Arvanitis, J. Minichiello, J.T. Price,

E.W. Thompson, Epidermal growth factor-induced

epithelio-mesenchymal transition in human breast carci-

noma cells, Lab. Invest. 83 (2003) 435–448.

[2] N.R. Alexander, N.L. Tran, H. Rekapally, C.E.

Summers, C. Glackin, R.L. Heimark, N-cadherin gene

expression in prostate carcinoma is modulated by

integrin-dependent nuclear translocation of Twist1,

Cancer Res. 66 (2006) 3365–3369.

[3] B.A. Alman, C. Li, M.E. Pajerski, S. Diaz-Cano,

H.J. Wolfe, Increased b-catenin and somatic APC

mutations in sporadic aggressive fibromatoses (desmoid

tumors), Am. J. Pathol. 151 (1997) 329–334.

[4] B.A. Alman, M.E. Pajerski, S. Diaz-Cano, K. Corboy,

H.J. Wolfe, Aggressive fibromatosis (desmoid tumor) is a

monoclonal disorder, Diagn. Mol. Pathol. 6 (1997)

98–101.

[5] M.F.C. Amary, P. Pauwels, E. Meulemans, G.M.

Roemen, L. Islam, B. Idowu, K. Bousdras, T.C. Diss,

P. O’Donnell, A.M. Flanagan, Detection of b-catenin

mutations in paraffin-embedded sporadic desmoid-type

fibromatosis by mutation-specific restriction enzyme

digestion (MSRED): an ancillary diagnostic tool, Am.

J. Surg. Pathol. 31 (2007) 1299–1309.

[6] F. Andre, B. Janssens, E. Bruyneel, F. van Roy,

C. Gespach, M. Mareel, M. Bracke, Alpha-catenin is

required for IGF-I-induced cellular migration but not

invasion in human colonic cancer cells, Oncogene 23

(2004) 1177–1186.

[7] L. Bertario, A. Russo, P. Sala, M. Eboli, M. Giarola,

F. D’amico, V. Gismondi, L. Varesco, M.A. Pierotti,

P. Radice, Genotype and phenotype factors as determi-

nants of desmoid tumors in patients with familial

adenomatous polyposis, Int. J. Cancer 95 (2001) 102–107.

[8] B. Bhattacharya, P. Dilworth, C. Iacobuzio-Donahue,

F. Ricci, K. Weber, M.A. Furlong, C. Fisher,

E. Montgomery, Nuclear b-catenin expression distin-

guishes deep fibromatosis from other benign and malig-

nant fibroblastic and myofibroblastic lesions, Am. J. Surg.

Pathol. 29 (2005) 653–659.

[9] D. Blanco, S. Vicent, E. Elizegi, I. Pino, M.F. Fraga,

M. Esteller, U. Saffiotti, F. Lecanda, L.M. Montuenga,

Altered expression of adhesion molecules and epithelial–

mesenchymal transition in silica-induced rat lung carci-

nogenesis, Lab. Invest. 84 (2004) 999–1012.

[10] J. Bohm, L. Niskanen, K. Kiraly, J. Kellokoski,

M. Eskelinen, S. Hollmen, E. Alhava, V.M. Kosma,

Expression and prognostic value of a-, b-, and g-cateninsin differentiated thyroid carcinoma, J. Clin. Endocrinol.

Metab. 85 (2000) 4806–4811.

[11] J.W. Carlson, C.D.M. Fletcher, Immunohistochemistry

for b-catenin in the differential diagnosis of spindle cell

lesions: analysis of a series and review of the literature,

Histopathology 51 (2007) 509–514.

[12] S.S. Cheon, A.Y. Cheah, S. Turley, P. Nadesan, R. Poon,

H. Clevers, B.A. Alman, b-catenin stabilization dysregu-

lates mesenchymal cell proliferation, motility, and inva-

siveness and causes aggressive fibromatosis and

hyperplastic cutaneous wounds, Proc. Natl. Acad. Sci.

USA 99 (2002) 6973–6978.

[13] A. Clairotte, I. Lascombe, S. Fauconnet, F. Mauny,

S. Felix, M.P. Algros, H. Bittard, B. Kantelip, Expression

of E-cadherin and a-, b-, g-catenins in patients with

bladder cancer, Am. J. Clin. Pathol. 125 (2006) 119–126.

[14] O. De Wever, W. Westbroek, A. Verloes, N. Bloemen,

M. Bracke, C. Gespach, E. Bruyneel, M. Mareel, Critical

role of N-cadherin in myofibroblast invasion and migra-

tion in vitro stimulated by colon-cancer-cell-derived

TGF-b or wounding, J. Cell Sci. 117 (2004) 4691–4703.

[15] P. Galiatsatos, W.D. Foulkes, Familial adenomatous

polyposis, Am. J. Gastroenterol. 101 (2006) 385–398.

[16] J.F. Gebert, C. Dupon, M. Kadmon, M. Hahn,

C. Herfarth, M. von Knebel Doeberitz, H.K. Schackert,

Combined molecular and clinical approaches for the

identification of families with familial adenomatous

polyposis coli, Ann. Surg. 229 (1999) 350–361.

[17] C. Gebert, J. Hardes, C. Kersting, C. August, H. Supper,

W. Winkelmann, H. Buerger, G. Gosheger, Expression of

b-catenin and p53 are prognostic factors in deep

aggressive fibromatosis, Histopathology 50 (2007) 491–497.

[18] M. Giarola, D. Wells, P. Mondini, S. Pilotti, P. Sala,

A. Azzarelli, L. Bertario, M.A. Pierotti, J.D. Delhanty,

P. Radice, Mutations of adenomatous polyposis coli

(APC) gene are uncommon in sporadic desmoid tumours,

Br. J. Surg. 78 (1998) 582–587.

[19] C.E. Gillet, D.W. Miles, K. Ryder, D. Skilton,

R.D. Liebman, R.J. Springall, D.M. Barnes,

A.M. Hanby, Retention of the expression of E-cadherin

and catenins is associated with shorter survival in grade

III ductal carcinoma of the breast, J. Pathol. 193 (2001)

433–441.

[20] J. Goldblum, J.A. Fletcher, Pathology and genetics of

tumors of soft tissue and bone, in: C.D.M. Fletcher,

K.K. Unni, F. Mertens (Eds.), World Health Organiza-

tion Classification of Tumors, IARC Press, Lyon, 2002,

pp. 83–84.

ARTICLE IN PRESST. Ferenc et al. / Pathology – Research and Practice 205 (2009) 311–324 323

[21] R.B. Hazan, R. Qiao, R. Keren, I. Badano, K. Suyama,

Cadherin switch in tumor progression, Ann. N. Y. Acad.

Sci. 1014 (2004) 155–163.

[22] R. Heymann, I. About, U. Lendahl, J.C. Franquin,

B. Obrink, T.A. Mitsiadis, E- and N-cadherin distribution

in developing and functional human teeth under normal

and pathological conditions, Am. J. Pathol. 160 (2002)

2123–2133.

[23] H.S. Hosalkar, E.J. Fox, T. Delaney, J.T. Torbert,

C.M. Ogilvie, R.D. Lackman, Desmoid tumors and

current status of management, Orthop. Clin. N. Am. 37

(2006) 53–63.

[24] M. Ilyas, I.P.M. Tomlinson, The interactions of APC,

E-cadherin and b-catenin in tumour development and

progression, J. Pathol. 182 (1997) 128–137.

[25] S. Islam, T.E. Carey, G.T. Wolf, M.J. Wheelock,

K.R. Johnson, Expression of N-cadherin by human

squamous carcinoma cells induces a scattered fibroblastic

phenotype with disrupted cell–cell adhesion, J. Cell Biol.

135 (1996) 1643–1654.

[26] B.V.S. Kallakury, C.E. Sheehan, E. Winn-Deen, J. Oliver,

H.A. Fisher, R.P. Kaufman Jr., J.S. Ross, Decreased

expression of catenins (a and b), p120 CTN, and

E-cadherin cell adhesion proteins and E-cadherin gene

promoter methylation in prostatic adenocarcinomas,

Cancer 92 (2001) 2786–2795.

[27] A. Kikuchi, Regulation of b-catenin signaling in the Wnt

pathway, Biochem. Biophys. Res. Commun. 268 (2000)

243–248.

[28] A.L. Knudsen, S. Bulow, Desmoid tumour in familial

adenomatous polyposis. A review of literature, Fam.

Cancer 1 (2001) 111–119.

[29] K.A. Knudsen, L. Myers, S.A. McElwee, A role for the

Ca2+-dependent adhesion molecule, N-cadherin, in myo-

blast interaction during myogenesis, Exp. Cell Res. 188

(1990) 175–184.

[30] A. Kovacs, J. Dhillon, R.A. Walker, Expression of

P-cadherin, but not E-cadherin or N-cadherin, relates to

pathological and functional differentiation of breast

carcinomas, J. Clin. Pathol.: Mol. Pathol. 56 (2003)

318–322.

[31] I. Lascombe, A. Clairotte, S. Fauconnet, S. Bernardini,

H. Wallerand, B. Kantelip, H. Bittard, N-cadherin as

a novel prognostic marker of progression in

superficial urothelial tumors, Clin. Cancer Res. 12

(2006) 2780–2787.

[32] C. Li, B. Bapat, B.A. Alman, Adenomatous polyposis coli

gene mutation alters proliferation through its b-catenin-regulatory function in aggressive fibromatosis (desmoid

tumor), Am. J. Pathol. 153 (1998) 709–714.

[33] A. Lugli, I. Zlobec, P. Minoo, K. Baker, L. Tornillo,

L. Terracciano, J.R. Jass, Prognostic significance of the

Wnt signaling pathway molecules APC, b-catenin and

E-cadherin in colorectal cancer – a tissue microarray-

based analysis, Histopathology 50 (2007) 453–464.

[34] B. Lustig, J. Behrens, The Wnt signaling pathway and its

role in tumor development, J. Cancer Res. Clin. Oncol.

129 (2003) 199–221.

[35] Y. Maeno, S. Moroi, H. Nagashima, T. Noda,

H. Shiozaki, M. Monden, S. Tsukita, A. Nagafuchi,

a-catenin-deficient F9 cells differentiate into signet ring

cells, Am. J. Pathol. 154 (1999) 1323–1328.

[36] C.A. Midgley, S. White, R. Howitt, V. Save,

M.G. Dunlop, P.A. Hall, D.P. Lane, A.H. Wyllie,

V.J. Bubb, APC expression in normal human tissues,

J. Pathol. 181 (1997) 426–433.

[37] M. Miettinen, J. Kopczynski, H.R. Makhlouf,

M. Sarlomo-Rikala, H. Gyorffy, A. Burke, L.H. Sobin,

J. Lasota, Gastrointestinal stromal tumors, intramural

leiomyomas, and leiomyosarcomas in the duodenum, Am.

J. Surg. Pathol. 27 (2003) 625–641.

[38] M. Miyaki, M. Konishi, R. Kikuchi-Yanoshita,

M. Enomoto, K. Tanaka, H. Takahashi, M. Muraoka,

T. Mori, F. Konishi, T. Iwama, Coexistence of somatic

and germ-line mutations of APC gene in desmoid tumors

from patients with familial adenomatous polyposis,

Cancer Res. 53 (1993) 5079–5082.

[39] E. Montgomery, M.S. Torbensen, M. Kaushal, C. Fisher,

S.C. Abraham, b-catenin immunohistochemistry sepa-

rates mesenteric fibromatosis from gastrointestinal stro-

mal tumor and sclerosing mesenteritis, Am. J. Surg.

Pathol. 26 (2002) 1296–1301.

[40] S. Nakajima, R. Doi, E. Toyoda, S. Tsuji, M. Wada,

M. Koizumi, S.S. Tulachan, D. Ito, K. Kami, T. Mori,

Y. Kawaguchi, K. Fujimoto, R. Hosotani, M. Imamur,

N-cadherin expression and epithelial–mesenchymal tran-

sition in pancreatic carcinoma, Clin. Cancer Res. 10

(2004) 4125–4133.

[41] T. Nakashima, C. Huang, D. Liu, K. Kameyama,

D. Masuya, S. Kobayashi, M. Kinoshita, H. Yokomise,

Neural-cadherin expression associated with angiogenesis

in non-small-cell lung cancer patients, Br. J. Cancer 88

(2003) 1727–1733.

[42] I.S. Nathke, The adenomatous polyposis coli protein,

J. Clin. Pathol.: Mol. Pathol. 52 (1999) 169–173.

[43] T.L. Ng, A.M. Gown, T.S. Barry, M.C. Cheang,

A.K. Chan, D.A. Turbin, F.D. Hsu, R.B. West,

T.O. Nielsen, Nuclear beta-catenin in mesenchymal

tumors, Mod. Pathol. 18 (2005) 68–74.

[44] G. Peterschulte, T. Lickfeld, G. Moslein, Das Desmoid-

problem, Chir. 71 (2000) 894–903.

[45] D. Rakheja, K.H. Molberg, C.A. Roberts, V.R. Jaiswal,

Immunohistochemical expression of b-catenin in solitary

fibrous tumors, Arch. Pathol. Lab. Med. 129 (2005)

776–779.

[46] A.S. Rocha, P. Soares, R. Seruca, V. Maximo, X. Matias-

Guiu, J. Cameselle-Teijeiro, M. Sobrinho-Simoes,Abnormalities of the E-cadherin/catenin adhesion com-

plex in classical papillary thyroid carcinoma and in its

diffuse sclerosing variant, J. Pathol. 194 (2001) 358–366.

[47] K.M. Ropponen, M.J. Eskelinen, P.K. Lipponen,

E.M. Alhava, V.M. Kosma, Reduced expression of acatenin is associated with poor prognosis in colorectal

carcinoma,

J. Clin. Pathol. 52 (1999) 10–16.

[48] T. Saito, Y. Oda, A. Sakamoto, S. Tamiya, N. Kinukawa,

K. Hayashi, Y. Iwamoto, M. Tsuneyoshi, Prognostic value

of the preserved expression of the E-cadherin and catenins

families of adhesion molecules and of b-catenin mutations

in synovial sarcoma, J. Pathol. 192 (2000) 342–350.


[49] T. Saito, Y. Oda, K. Tanaka, S. Matsuda, S. Tamiya,

Y. Iwamoto, M. Tsuneyoshi, b-catenin nuclear expression

correlates with cyclin D1 overexpression in sporadic

desmoid tumours, J. Pathol. 195 (2001) 222–228.

[50] K. Shitoh, F. Konishi, T. Iijima, T. Ohdaira, K. Sakai,

K. Kanazawa, M. Miyaki, A novel case of a sporadic

desmoid tumour with mutation of the b-catenin gene,

J. Clin. Pathol. 52 (1999) 695–696.

[51] N.J.H. Strut, S.K. Clark, Current ideas in desmoid

tumours, Fam. Cancer 5 (2006) 275–285.

[52] S. Tejpar, C. Li, C. Yu, R. Poon, H. Denys, R. Sciot,

E. Van Cutsem, J.J. Cassiman, B.A. Alman, Tcf-3

expression and b-catenin mediated transcriptional activa-

tion in aggressive fibromatosis (desmoid tumour),

Br. J. Cancer 85 (2001) 98–101.

[53] R.A. Walker, Quantification of immunohisto-

chemistry – issues concerning methods, utility and

semiquantitative assessment I, Histopathology 49 (2006)

406–410.

[54] S.W. Weiss, J.R. Goldblum, Enzinger and Weiss’s Soft

Tissue Tumors, fourth ed., St. Luis, Mosby, 2001,

pp. 309–346.

[55] N.A. Wong, M. Pignatelli, b-catenin – a linchpin in

colorectal carcinogenesis?, Am. J. Pathol. 160 (2002)

389–401.

[56] J. Yu, M.P.A. Ebert, S. Miehlke, H. Rost, U. Lendeckel,

A. Leodolter, M. Stolte, E. Bayerdorffer, P. Malferthei-

ner, a-catenin expression is decreased in human gastric

cancers and in the gastric mucosa of first degree relatives,

Gut 46 (2000) 639–644.

Documents

Analysis of APC, α-, β-catenins, and N-cadherin protein expression in aggressive fibromatosis (desmoid tumor)