cyclin-d1 and Fibromatosis--Aggressive

Sporadic desmoid-type fibromatosis (DTF) is a rare soft tissue tumor of mesenchymal origin. It is characterized by local invasive growth and unpredictable growth behavior. Three distinct mutations involving the CTNNB1 (β-catenin) gene have been identified in the vast majority of DTF tumors, which cause activation of the Wnt signaling pathway and impact prognosis. This study examines whether the different CTNNB1 mutants (T41A, S45F) occurring in DTF tumors differentially affect Wnt signaling activity, which might explain the different disease course between DTF patients harboring different CTNNB1 mutations.. Real-time polymerase chain reaction (RT-PCR) on 61 formalin fixed paraffin embedded DTF samples with known CTNNB1 status was used to measure the relative mRNA expression level of Wnt target genes AXIN2, DKK1 and CCND1. Additionally, publicly available mRNA expression data retrieved from the Gene Expression Omnibus of 128 DTF samples were used for an unsupervised cluster analyses based on the expression of a selection of Wnt targets.. No statistically significant difference in relative expression levels of Wnt target genes AXIN2, DKK1 and CCND1 was identified between either CTNNB1 wild-type, S45F or T41A mutated DTF samples. Moreover, the hierarchical cluster analyses using selected Wnt targets did not discriminate between different CTNNB1 mutation types.. No differences in the expression levels of Wnt target genes were observed between the different CTNNB1 mutation types in DTF tumors. Further studies are needed to decipher the mechanism accounting for the diverse disease courses between DTF patients with different CTNNB1 variants.

Topics: Adolescent; Adult; Aged; Axin Protein; beta Catenin; Child; Child, Preschool; Cyclin D1; Female; Fibromatosis, Abdominal; Fibromatosis, Aggressive; Gene Expression; Humans; Infant; Intercellular Signaling Peptides and Proteins; Male; Middle Aged; Mutation; RNA, Messenger; Soft Tissue Neoplasms; Wnt Signaling Pathway; Young Adult

Estrogen receptor signaling and cyclin D1 have a major role in tumor cell proliferation in breast cancer. Desmoid tumors are rare neoplasms that may respond to endocrine treatment. The present study aimed to investigate the expression levels and the clinical relevance of estrogen receptor beta (ERβ) and cyclin D1 in desmoid tumors.. This study consists of 83 patients with a surgically treated desmoid tumor. ERβ and cyclin D1 expression was examined by immunohistochemistry in tissue microarrays. Cyclin A and Ki67 were studied in our previous work.. Median ERβ expression was 10.8%. ERβ expression correlated with expression of the proliferation antigens Ki67 (r. ERβ and cyclin D1 immunopositivity correlated with high proliferation in desmoid tumors. High ERβ expression might be predictive for postoperative recurrence.

Topics: Adult; Biomarkers, Tumor; Cell Growth Processes; Cyclin D1; Estrogen Receptor beta; Female; Fibromatosis, Aggressive; Humans; Immunohistochemistry; Male; Neoplasm Recurrence, Local; Predictive Value of Tests; Retrospective Studies; Tissue Array Analysis

Desmoid tumors are benign mesenchymal neoplasms with a locally aggressive nature. The mutational status of β-catenin gene (CTNNB1) is presumed to affect the tumorous activity of the cells. In this study, we isolated three kinds of desmoid cell with different CTNNB1 status, and compared their characteristics. Cells were isolated from three patients with abdominal wall desmoid during surgery, all of which were resistant to meloxicam treatment. The mutational status of the CTNNB1 exon 3 was determined for both parental tumor tissues and isolated cultured cells. β-catenin expression was determined with immunohistochemistry. Responsiveness to meloxicam was investigated with MTS assay together with COX-2 immunostaining. mRNA expressions of downstream molecules of Wnt/β-catenin pathway were determined with real-time RT-PCR. Three kinds of cell isolated from desmoid tumors harboring different CTNNB1 mutation status (wild type, T41A, and S45F), all exhibited a spindle shape. These isolated cells could be cultured until the 20th passage with unchanged proliferative activity. Nuclear accumulation of β-catenin was observed in all cultured cells, particularly in those with S45F. Proliferating activity was significantly suppressed by meloxicam (25 μmol/L, P < 0.007) in all three cell cultures, of which parental desmoid was resistant to meloxicam clinically. The mRNA expressions of Axin2, c-Myc, and Cyclin D1 differently increased in the three cultured cell types as compared with those in human skin fibroblast cells (HDF). Inhibitors of Wnt/β-catenin pathway downregulated Axin2, c-Myc, and Cyclin D1 significantly. Isolated and cultured desmoid tumor cells harboring any one of the CTNNB1 mutation status had unique characteristics, and could be useful to investigate desmoid tumors with different mutation status of CTNNB1.

Topics: Adult; Antineoplastic Agents; Apoptosis; Axin Protein; beta Catenin; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclooxygenase 2; DNA Mutational Analysis; Fibromatosis, Aggressive; Gene Expression; Gene Expression Regulation, Neoplastic; Genes, myc; Humans; Meloxicam; Mutation; RNA, Messenger; Thiazines; Thiazoles; Tumor Cells, Cultured; Wnt Signaling Pathway; Young Adult

Desmoid tumors (DTs), the commonest extra-intestinal manifestation of familial adenomatosis polyposis (FAP), are monoclonal neoplasms demonstrating fibroblastic - myofibroblastic differentiation; they are locally invasive without metastatic capacity. FAP-associated DT natural history knowledge is limited; we examined patient and tumor characteristics for a FAP-DT cohort and evaluated anti-DT therapy molecular target expression levels (immunohistochemical analyses, FAP-DT tissue microarray; TMA). Forty-four patients were classified as intra-abdominal (IA; n=26), abdominal wall (AW)/extra-abdominal (EA; n=12) or concomitant IA/AW (n=6) based on DT primary diagnosis location. Positive family histories were found in 62% of FAP versus 10% of DT patients. Surgery was the mainstay therapy for AW/EW patients, whereas IA DTs received surgery, chemotherapy, radiotherapy, tamoxifen, NSAIDs, and/or imatinib. Eight of 20 completely resected DTs in the IA and AW/EA groups recurred; 12 of 38 patients in these groups (33%) developed secondary lesions elsewhere. Two intestinal mesenteric DT patients died of disease, three from other cancers, 27 are alive with disease and 12 are alive without disease. All evaluable FAP-DT exhibited nuclear β-catenin, 65% were positive for cyclin D1, and 66% expressed nuclear p53. No ERα expression was observed, but ERβ was expressed in 72%. COX2 was expressed in all evaluable FAP-DTs. KIT was rarely found in DTs but both PDGFRs and their ligands were expressed. Comparing biomarker expression (IA vs. EA DTs), only nuclear ER-ß staining was significantly higher in EA lesions (p=0.0070); no other markers were site informative. Enhanced knowledge of FAP-DT molecular underpinnings will facilitate development of novel therapeutic strategies.

Topics: Abdominal Neoplasms; Adenomatous Polyposis Coli; Adolescent; Adult; beta Catenin; Biomarkers, Tumor; Cohort Studies; Cyclin D1; Cyclooxygenase 2; Estrogen Receptor alpha; Estrogen Receptor beta; Female; Fibromatosis, Aggressive; Humans; Immunohistochemistry; Male; Middle Aged; Platelet-Derived Growth Factor; Proto-Oncogene Proteins c-kit; Receptors, Platelet-Derived Growth Factor; Retrospective Studies; Tissue Array Analysis; Tumor Suppressor Protein p53; Young Adult

To explore the role of abnormalities of chromosome 8, APC and beta-catenin genes in tumorigenesis of aggressive fibromatosis.. Trisomy 8 was detected by interphase fluorescence in situ hybridization (FISH). The APC gene and beta-catenin gene mutations were detected by denaturing high performance liquid chromatography (DHPLC) and direct sequence analysis after the PCR transition.. The rate of trisomy 8 in recurrent tumors (62.5%, 5/8) was significantly higher than that in the primary tumors (8.3%, 1/12). Somatic substitution of APC gene was found in 18 of 69 (26.1%) aggressive fibrometases. Somatic transition of beta-catenin gene was detected in 13 of 69 (18.8%) and mutation at codon 41 in exon 3 involving threonine residues implicated in the degradation of beta-catenin. The abnormal expression of beta-catenin had no significant correlation with the mutation of APC or beta-catenin gene. The group with positively expressed beta-catenin protein showed a significant higher c-myc protein expression than those without (P = 0.001). The Ki-67 index was extremely low in all the lesions. The apoptosis index (AI) of the groups with positively expressed c-myc and cyclin D1 showed significantly lower AI than those without.. Trisomy 8 may serve as a useful predictor of recurrence in aggressive fibromatosis. There are somatic mutations of the APC and beta-catenin genes in the aggressive fibromatosis, and there are abnormalities in the Wnt signaling pathway. These abnormalities may result in the aberrances of cell proliferation and apoptosis, which are likely to be import factors in the tumorigenesis.

Topics: Adenomatous Polyposis Coli Protein; Apoptosis; beta Catenin; Chromosomes, Human, Pair 8; Cyclin D1; Fibromatosis, Aggressive; Genes, APC; Humans; Ki-67 Antigen; Neoplasm Recurrence, Local; Point Mutation; Proto-Oncogene Proteins c-myc; Signal Transduction; Trisomy; Wnt Proteins

The abnormalities of the Wnt signalling pathway in desmoid-type fibromatosis were analysed, with the purpose of exploring the mechanism of tumorigenesis and progression.. The clinical and histopathological features of 96 cases were analysed. Beta-catenin, cyclin-D1, c-myc, and Ki-67 proteins were detected in 69 cases using formalin-fixed, paraffin-embedded tissues. Using the same materials, apoptosis of the tumour cells was investigated by terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling (TUNEL) testing. Polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) assay, and sequencing were performed to detect abnormalities of the adenomatous polyposis coli (APC) and beta-catenin genes.. APC gene mutations were found in 18 cases (26.1%, 18/69). Somatic mutations of codon 41 in exon 3 of beta-catenin were detected in 13 cases (18.8%, 13/69). No correlation of beta-catenin abnormal expression with the mutations of APC gene or beta-catenin gene was identified (p>0.05). The cases with abnormal beta-catenin expression showed a higher level of c-myc protein expression (69.7%, 23/33) than those without (22.2%, 8/36, p = 0.001). The apoptotic indices (AIs) were significantly lower in cyclin-D1 positive cases and c-myc positive cases (p = 0.015, p = 0.007).. There are somatic mutations of the APC and beta-catenin gene in desmoid-type fibromatosis, and there are abnormalities in the Wnt signalling pathway. These abnormalities may result in aberrant cell proliferation and apoptosis, which are likely to be important factors in tumorigenesis and progression.

Topics: Apoptosis; Base Sequence; beta Catenin; Chromatography, High Pressure Liquid; Cyclin D1; Fibromatosis, Aggressive; Genes, APC; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Ki-67 Antigen; Mutation; Polymerase Chain Reaction; Proto-Oncogene Proteins c-myc; Signal Transduction; Wnt Proteins

We report a case of intrathoracic desmoid tumor without familial adenomatous polyposis and demonstrate beta-catenin mutation of exon 3. A 15-year-old male presented with a desmoid tumor after having sustained an assault. In an examination for a mutation of the beta-catenin gene, an activating mutation from ACC (Thr) to GCC (Ala) at codon 41 was found. Immunohistochemical staining showed that accumulated beta-catenin protein was predominantly localized in the nuclei of desmoid cells, and cyclin D1 protein was also overexpressed. These findings might suggest that an activating mutation of the beta-catenin gene affected regulation of the cyclin D1 gene, resulting in the generation of intrathoracic sporadic desmoid tumor, which arose at the site of posttraumatic injury.

Topics: Adolescent; beta Catenin; Cyclin D1; Exons; Fibromatosis, Aggressive; Gene Expression Regulation; Humans; Male; Mutation; Thoracic Neoplasms; Tomography, X-Ray Computed

Desmoid tumors arising in the lung and pleura are extremely rare and can resemble other, more common neoplasms native to these sites. Alterations of the adenomatous polyposis coli/beta-catenin pathway have been detected in sporadic desmoid tumors and have been associated with nuclear accumulation of beta-catenin and overexpression of cyclin D1.. To analyze the expression of beta-catenin and cyclin D1 in desmoid tumors and solitary fibrous tumors (SFTs), and to compare the utilities of these substances for distinguishing between these entities with those of other, more commonly used stains.. Formalin-fixed, paraffin-embedded sections of 4 desmoid tumors (1 pulmonary, 1 pleural, 2 pleural/chest wall), and 5 benign and 6 malignant SFTs of the pleura were immunostained for beta-catenin, cyclin D1, ALK1, CD34, vimentin, desmin, smooth muscle actin, muscle-specific actin, S100, and pancytokeratin. Staining intensity and the percentage of stained tumor cells were assessed semiquantitatively.. Diffuse moderate or strong nuclear staining for beta-catenin was found in all desmoid tumors, 4 of 5 benign SFTs, and 2 of 6 malignant SFTs. All cases except 1 benign SFT showed concurrent cytoplasmic staining. Nuclear and cytoplasmic cyclin D1 staining was increased in all groups. The best distinction between desmoid tumors and SFTs was provided by CD34 (desmoid tumors, 0/4; SFTs, 8/11) and smooth muscle actin (desmoid tumors, 4/4; SFTs, 0/11).. Our findings suggest that alterations in the adenomatous polyposis coli/beta-catenin pathway and cyclin D1 dysregulation may contribute to the pathogenesis of pleuropulmonary desmoid tumors and SFTs. CD34 and smooth muscle actin stains are particularly useful for differentiating between pleuropulmonary desmoid tumors and SFTs.

Topics: Actins; Adult; Aged; Antigens, CD34; beta Catenin; Cell Nucleus; Child, Preschool; Cyclin D1; Cytoplasm; Diagnosis, Differential; Female; Fibromatosis, Aggressive; Humans; Immunohistochemistry; Leiomyoma; Lung Neoplasms; Male; Middle Aged; Muscle, Smooth; Pleural Neoplasms; Staining and Labeling

We screened for genetic alterations of adenomatous polyposis coli (APC) and beta-catenin genes in 17 frozen specimens from 12 cases of sporadic desmoid tumors and then subdivided these cases into two groups according to the results of mutational analysis. We further examined mRNA expression of beta-catenin and cyclin D1 by TaqMan PCR and compared the mRNA expression within both groups. Single-strand conformation polymorphism analysis followed by DNA direct sequencing revealed beta-catenin mutation in 3 of 12 cases (6 of 17 specimens), whereas no APC missense mutations in the mutation cluster region were found. TaqMan PCR revealed extremely higher mRNA expression of beta-catenin and cyclin D1 in desmoid tumors, compared with those of normal skeletal muscles. In the beta-catenin mutated group, cyclin D1 mRNA expression was significantly higher than that of the beta-catenin wild-type group (p = 0.0120, Mann-Whitney U test). In addition, in the beta-catenin mutated group, beta-catenin mRNA expression was also significantly higher than that of the beta-catenin wild-type group (p = 0.0036, Mann-Whitney U test). All cases of desmoid tumors showed detectable beta-catenin nuclear expression immunohistochemically. These results suggest that a continuously elevated beta-catenin protein level caused by the beta-catenin mutation itself may have a stronger power that can transactivate transcription in vivo. Furthermore, the results provide a possible association between higher beta-catenin mRNA expression and mutated beta-catenin in sporadic desmoid tumors. This may suggest that the beta-catenin gene may be up-regulated by mutated or continuously elevated beta-catenin protein, that is, the beta-catenin gene may also be one of the targeted genes in the APC-beta-catenin-Tcf pathway.

Topics: Adolescent; Adult; beta Catenin; Child; Cyclin D1; Cytoskeletal Proteins; Disease-Free Survival; Female; Fibromatosis, Aggressive; Gene Expression Regulation, Neoplastic; Genes, APC; Humans; Kinetics; Male; Middle Aged; Mutation; Polymerase Chain Reaction; Recurrence; RNA, Messenger; Taq Polymerase; Time Factors; Trans-Activators; Transcription, Genetic

The immunohistochemical expression of beta-catenin, cyclin D1, Ki-67 and PCNA was Examined in 38 cases of sporadic extra-abdominal or abdominal-wall desmoid tumours without familial adenomatous polyposis (FAP), to evaluate the hypothesis that the accumulated beta-catenin within the nuclei could affect the regulation of the cyclin D1 gene. There was a statistically significant correlation between beta-catenin accumulation and cyclin D1 overexpression (p=0.029). Each group with beta-catenin accumulation or cyclin D1 overexpression showed a higher PCNA-LI than those without, the difference being statistically significant (p=0.007, p=0.004, respectively). Differential PCR was also performed to detect amplification of the cyclin D1 gene and mutational analysis was undertaken for exon 3 of the beta-catenin gene. Amplification of the cyclin D1 gene was observed in 13 out of 22 cases (59.1%). There were nine-point mutations in 7 out of 18 cases (38.9%). The distribution of beta-catenin mutation fell within a wide range, from codon 21 to codon 67. In conclusion, beta-catenin nuclear expression correlated with cyclin D1 overexpression in sporadic desmoid tumours, which could be an in vivo model system for the APC-beta-catenin-Tcf pathway. In addition, beta-catenin mutations in desmoid tumours occurred at an unusually wide range of sites within the gene.

Topics: Adolescent; Adult; Aged; beta Catenin; Cell Nucleus; Child; Cyclin D1; Cytoskeletal Proteins; Female; Fibromatosis, Abdominal; Fibromatosis, Aggressive; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Mutation, Missense; Neoplasm Proteins; Proliferating Cell Nuclear Antigen; Trans-Activators