Skip to main content

Gene Expression Profiling in Familial Adenomatous Polyposis Adenomas and Desmoid Disease

Abstract

Gene expression profiling is a powerful method by which alterations in gene expression can be interrogated in a single experiment. The disease familial adenomatous polyposis (FAP) is associated with germline mutations in the APC gene, which result in aberrant β-catenin control. The molecular mechanisms underlying colorectal cancer development in FAP are being characterised but limited information is available about other symptoms that occur in this disorder. Although extremely rare in the general population, desmoid tumours in approximately 10% of FAP patients. The aim of this study was to determine the similarities and differences in gene expression profiles in adenomas and compare them to those observed in desmoid tumours. Illumina whole genome gene expression BeadChips were used to measure gene expression in FAP adenomas and desmoid tumours. Similarities between gene expression profiles and mechanisms important in regulating formation of FAP adenomas and desmoid tumours were identified. This study furthers our understanding of the mechanisms underlying FAP and desmoid tumour formation.

Introduction

Familial adenomatous polyposis (FAP) is a rare form of colorectal cancer caused by germline mutations in the adenomatous polyposis coli (APC) gene. Approximately 70–90% of FAP patients have identifiable germline mutations in APC [1, 2]. FAP is clinically characterized by the formation of hundreds to thousands of adenomas that carpet the entire colon and rectum [3]. Although initially benign the risk of malignant transformation increases with age such that, if left untreated, colorectal carcinoma usually develops before the age of 40 years [4].

Loss of APC results in dysregulation of the Wnt signalling pathway that leads to the constitutional activation of the transcription factor Tcf-4, which has been associated with adenoma formation [5]. Alterations in Wnt signalling cause stem cells to retain their ability to divide in the upper intestinal crypt, thereby forming monocryptal adenomas [6]. Eventually the adenomas may acquire metastatic potential, resulting in carcinoma development [7]. Not all adenomas will progress to malignant tumours; however, due to the abundance of adenomas carcinoma development is virtually assured [8].

Apart from the apparent loss of APC function, little is known about the molecular processes involved in adenoma initiation [6]. Similarly, the molecular events occurring during the transformation of adenomas into carcinomas are poorly understood, as are the mechanisms that underlie the development of extra-colonic disease in FAP.

It is well established that FAP patients are susceptible to benign extra-colonic tumours, including desmoid tumours [3]. Although rare in the general population, desmoids occur in approximately 10% of FAP patients and they are the second most common cause of death [9]. Desmoid tumours are poorly encapsulated and consist of spindle-shaped fibroblast cells with varying quantities of collagen [10]. Despite their apparent inability to metastasize, desmoid tumours can be extremely aggressive [11].

It has been speculated that desmoid formation is a result of an abnormal wound healing response [12]. Desmoids can affect surrounding viscera, causing potentially fatal complications [13]. FAP-associated desmoid tumours are usually associated with germline APC mutations [14], but somatic APC mutations have been detected in sporadic desmoid tumours [15].

Microarray technology has an enormous potential for applications in the endeavour to better understand tumours and their development [16]. The ability to detect expression levels of thousands of genes can identify particular genes that are either up- or down-regulated in different tumour types [17]. Tumours that are currently categorized by similar morphology, such as desmoid tumours, may be more usefully divided into subtypes according to their expression profiles [18]. Particular expression profiles in tumours may also be capable of predicting the clinical outcome in specific patients in the early stages of tumour development [18]. In colorectal cancer, gene expression profiles of adenomas and adenocarcinomas have been compared and subsets of genes expressed at common levels in both lesions have been identified as well as expression patterns that are unique to each [19]. Gene expression profiling has the potential to identify factors involved in the malignant transformation of adenomas, and may aid in the diagnosis of benign versus malignant disease.

Although genome-wide expression studies have been reported on FAP adenomas and desmoid tumours, the present one of the first to compare the two tissue types. The first aim of this study was to identify distinct gene expression profiles for colorectal and stomach FAP adenomas and desmoid tumours. The second aim was to determine the similarity between the gene expression profiles in FAP adenomas and desmoid tumours to identify mechanisms important in regulating formation of these lesions. To achieve this, mRNA from normal colon, FAP stomach and colon adenomas and desmoid tumours was measured using whole human genome expression BeadChips (Illumina). The findings of this study further our understanding of the mechanisms underlying FAP and desmoid tumour formation.

Materials and methods

FAP adenoma and tumour tissue and controls

Frozen adenoma tissue from 4 FAP patients was available for this study. Colorectal FAP adenoma A was from an individual aged 40 at the time of surgery. Genetic testing revealed a heterozygous A5465T change in the APC gene, causing a missense change from aspartic acid to valine at position 1822 in the amino acid sequence. The specimen obtained for this study was obtained as a result of a proctocolectomy. The pathology report indicated that over 100 tubulovillous adenomas were present in the original specimen, with no evidence of invasive tumour. Patients B, C and D harboured the same frameshift mutation, a 4 base pair deletion at position 3462–3465 of the APC gene. Patient B was diagnosed with FAP at the age of 11 years, patient C at 13 years of age, and patient D at the age of 37 years. One gastric adenoma was obtained from patient D, in addition to a colonic adenoma. Normal colon tissue from 7 healthy individuals with no history of FAP or desmoid disease was used as a mixed reference sample for this study.

Desmoid Disease Tissue

Desmoid tumour tissue from two individuals was available for this study. Patient A had FAP-associated desmoid disease. There was a family history of FAP, but no known history of desmoid disease. The individual harboured a 1 bp deletion in exon 15 of the APC gene resulting in a frameshift that introduced a premature stop codon at amino acid position 964. Patient B had a family history of FAP and desmoid disease. This patient harboured a 17 bp duplication in exon 15 of the APC gene, which introduced a premature stop codon at amino acid position 1969. A previously established fibroblast cell line from a healthy individual with no history of FAP or desmoid disease was used as a control for this study. The fibroblast cell line was cultured in 1× Complete DMEM media at 37°C (5% CO2).

RNA Extraction

2–3 mm2 pieces of fresh frozen FAP adenoma and desmoid tumour tissue were cut from the original sample and transferred immediately to 1 ml Trizol reagent (Invitrogen, USA). Similarly, approximately 1–10 × 106 control fibroblast cells were lysed in 1 ml Trizol reagent (Invitrogen, USA). RNA was extracted per manufacturer's instructions. The RNA pellet was washed with 75% ethanol, before being dissolved in 20 μl water.

The total RNA was purified using a Qiagen RNeasy MiniElute Cleanup Kit as per manufacturer's instructions. The concentration of the purified total RNA samples was measured using a Quant-It RiboGreen RNA Assay Kit (Invitrogen, USA) and a fluorometer (Fluostar OPTIMA) as per manufacturer's instructions.

RNA amplification

To synthesise first and second strand cDNA and amplify biotinylated cRNA from the total RNA, an Illumina Totalprep RNA Amplification Kit was used as per manufacturer's instructions.

The purified cRNA samples were quantified to determine the volume required for the BeadChip hybridisation step via the Quant-iT RiboGreen RNA Assay Kit as described previously.

Illumina BeadChip Procedure

Hybridisation to the Illumina Sentrix 8 BeadChip was performed according to the manufacturer's instructions without modification. The Sentrix 8 BeadChips were read using an Illumina Beadarray reader (San Diego, CA, USA).

Data Analysis

Analysis and normalisation of expression data from the 24,000 transcripts was carried out using BeadStudio 2.0 (Illumina, San Diego, CA, USA). The t-test error model and cubic spline normalisation was used for all samples. A differential analysis was applied to all adenoma and tumour samples using the Illumina custom test of significance, utilising the mixed normal colon control as the reference group. GeneSpring 5.0 (Agilant, Santa Clara, CA, USA) used standard correlation and distance to create dendrograms (Experiment trees) to show relationships between gene expression profiles. A second dendrogram (Gene tree) was created for each gene list using standard correlation and distance to show relationships between the expression levels of genes across the groups.

Results

Gene expression data from over 23,000 genes on Illumina HumRef-8 BeadChips was analysed and normalised using Illumina BeadStudio 2.0 software. Cubic spline normalisation and the t-test error model were employed for all the FAP adenoma, normal colon and desmoid tumour samples. Correlation analyses identified the average R2 value of the duplicates for each sample as 0.950 ± 0.04. An average of each duplicate pair was then taken before additional analysis was carried out.

Differential gene expression analysis in FAP adenomas and healthy colon tissue

Differential analysis using the mixed normal colon control as the reference group was applied to all adenoma and tumour samples. Genes in each analysis were excluded if their fluorescence detection score was less than 0.99, and if their differential score was less than 13 (p > 0.05). From the genes that met the exclusion criteria, according to detection and differential scores, lists were generated for genes both up- and down-regulated more than 2-fold in the FAP adenoma samples compared to the mixed normal colon control. The genes commonly up- and down-regulated across all the FAP adenomas are shown in Tables 1 and 2 and genes that were commonly up- or down-regulated across the 4 colorectal FAP adenomas only are shown in Tables 3 and 4 respectively.

Table 1 Genes commonly up-regulated more than 2-fold in all FAP polyps compared to normal colon
Table 2 Genes commonly down-regulated more than 2-fold in all FAP polyps compared to normal colon
Table 3 Genes commonly up-regulated 2-fold or more in colorectal FAP polyps compared to normal colon
Table 4 Genes commonly down-regulated 2-fold or more in colorectal FAP polyps compared to normal colon

Cluster analysis was performed using GeneSpring 5.0 software in order to further characterise the similarity across the FAP samples and to determine if there was differential gene expression compared to healthy colon tissue. The stomach FAP duplicates display profiles slightly distinct from the other FAP adenomas. The normal colon duplicate profiles are unique to all other profiles (Figure 1).

Figure 1
figure 1

Cluster analysis of FAP polyps and mixed normal colon. The columns represent the gene expression profiles of each sample. Green – low expression level, yellow – medium expression level, red – high expression level. The relationships between each sample are shown by the upper dendrogram. The colouring in the upper dendrogram represents the sample type: green (left) – normal colon; blue – colorectal FAP polyps; yellow – stomach FAP. 1 – Normal Colon Duplicate; 2 – Normal Colon Duplicate; 3 – Colorectal FAP Polyp A Duplicate; 4 – Colorectal FAP Polyp A Duplicate; 5 – Colorectal FAP Polyp D Duplicate; 6 – Colorectal FAP Polyp D Duplicate; 7 – Colorectal FAP Polyp B Duplicate; 8 – Colorectal FAP Polyp B Duplicate; 9 – Colorectal FAP Polyp C Duplicate; 10 – Colorectal FAP Polyp C Duplicate; 11 – Stomach FAP Polyp D Duplicate; 12 – Stomach FAP Polyp D Duplicate.

Differential gene expression analysis in desmoid tumours and control fibroblasts

The average expression in the desmoid tumours was compared to the control fibroblast cell line and significantly altered expression identified by differential gene expression analysis. Genes in each analysis were excluded if their fluorescence detection score was less than 0.99, and if their differential score was less than 13 (p > 0.05). Genes with differential expression and up- or down-regulated more than 2-fold in the desmoid tumour samples compared to the normal fibroblast cell line were compiled into lists (Tables 5 and 6).

Table 5 Genes commonly up-regulated 2-fold or more in desmoid tumours compared to normal fibroblast cells
Table 6 Genes commonly down-regulated 2-fold or more in desmoid tumours compared to normal fibroblast cells

To reveal any correlation between the expression profiles of desmoid tumours and FAP adenomas, the data from each group were compared. In the upper dendrogram (Figure 2) it can be seen that all the FAP adenomas cluster in the same group. The desmoid tumours and the normal fibroblast cell line clustered in an entirely different group to the FAP samples. The FAP adenomas and the normal colon have distinct gene profiles compared to the desmoid tumours and the normal fibroblasts. Within the FAP adenomas, the stomach adenoma and the normal colon have slightly different gene profiles compared to the colorectal adenomas.

Figure 2
figure 2

Cluster analysis of FAP polyps, normal colon, desmoid tumours and normal fibroblasts. The columns represent the gene expression profiles of each sample. Green – low expression level, yellow – medium expression level, red -high expression level. The relationships between each sample are shown by the upper dendrogram. The colouring in the upper dendrogram represents the sample type: green (left) – normal colon; blue – colorectal FAP polyps; orange – stomach FAP polyp; green (right) – desmoid tumours; purple – fibroblast cell line. 1 – Normal Colon Duplicate; 2 – Normal Colon Duplicate; 3 – Colorectal FAP Polyp A Duplicate; 4 – Colorectal FAP Polyp A Duplicate; 5 – Colorectal FAP Polyp D Duplicate; 6 – Colorectal FAP Polyp D Duplicate; 7 – Colorectal FAP Polyp B Duplicate; 8 – Colorectal FAP Polyp B Duplicate; 9 – Colorectal FAP Polyp C Duplicate; 10 – Colorectal FAP Polyp C Duplicate; 11 – Stomach FAP Polyp D Duplicate; 12 – Stomach FAP Polyp D Duplicate; 13 – Desmoid Tumour A Duplicate; 14 – Desmoid Tumour A Duplicate; 15 – Desmoid Tumour C Duplicate; 16 – Desmoid Tumour C Duplicate; 17 – Fibroblast Cell Line Duplicate; 18 – Fibroblast Cell Line Duplicate.

Discussion

In this study, 24 K Illumina HumRef-8 BeadArrays were used to compare gene expression of FAP adenomas, desmoid tumours and normal fibroblasts. To date there have been a number of small scale gene expression studies on FAP adenoma tissue, the vast majority of which have employed immunohistochemistry (IHC). Most of these studies have been performed on individual genes that include E-cadhein, α-, β- and -catenin, COX-1, COX-2, and c-myc [20–25]. In addition, one study used semi-quantitative RT-PCR to study GKLF [26]. The only report examining global gene expression in human FAP adenoma tissue identified 84 differentially expressed genes in adenomas compared to normal colon tissue [27].

In this study, the gene expression profiles obtained from the FAP adenomas indicate that colorectal adenomas are similar but distinctly different to the stomach adenomas. There were a large number of commonly expressed genes identified across the colorectal FAP adenomas, but when the differentially expressed genes from the stomach adenoma were included in the analysis the number of commonly expressed genes decreased dramatically. The genes that were differentially expressed in the four colonic adenomas and one stomach adenoma were investigated more closely in an attempt to identify common genetic features in FAP. From this analysis genes involved in the cell cycle, transcription and metabolism were the most frequently up-regulated. The most frequently down-regulated genes included those involved in metabolism, cell adhesion, signal transduction, transcription and transport. Since adenomas develop due to a breakdown in the fidelity of the Wnt signalling pathway it was not surprising to observe the over-expression of genes involved in cell cycle progression.

Altered Expression of Wnt/β-catenin Target Genes in Colorectal FAP Adenomas

It has been long established that deregulation of the Wnt signalling pathway due to APC mutations plays a major role in the progression of FAP [5]. The Wnt/β-catenin signalling pathway is involved in the control of expression of Sox9, PTTG1 and EphB2, all of which were found to be up-regulated by more than 2-fold in all the colorectal FAP adenomas compared to the normal colon.

PTTG1 is regulated by a TCF binding sequence in its promoter region [28]. The normal function of PTTG1 is to regulate chromosome segregation during cell division [29]. Over-expression of PTTG1 has been reported frequently in various types of cancer, including colorectal, and has been associated with angiogenesis [30–32]. The role of PTTG1 in angiogenesis is thought to be a result of its part in mediating the secretion of the basic fibroblast growth factor into the extracellular matrix, which promotes proliferation and migration of colorectal cancer cells [30, 31].

The Sox9 gene encodes a transcription factor that is required for chondrogenesis and male gonad development [32], which is under the control of the Wnt signalling pathway [33]. The expression of the Sox9 gene in the intestine is dependent on the activity of the β-catenin/TCF-4 complex, although it is unknown whether this complex interacts directly with the Sox9 promoter or through another of its targets [33].

The EphB2 gene encodes the Eph receptor B, which has been shown to be a target of the Wnt signalling pathway [34]. There is evidence to suggest that normal patterning in the epithelium of the intestinal crypts is coordinated by EphB2 and its ligand, ephrin B [34]. Over-expression of EphB2 is often found in colorectal cancers, but there is confusion about its role in tumourigenesis. Many studies on other tumours have reported EphB2 over-expression as a marker of poor prognosis, but recent studies in colorectal cancer have suggested otherwise [35, 36].

Altered Expression of Cell Cycle-Related Genes in Colorectal FAP Adenomas

A number of genes found to be commonly up-regulated in the adenomas used in this study have previously been reported as being over-expressed in various types of cancers. These genes include the cell cycle-related genes Chromosome condensation protein G (HCAP-G), Protein regulator of cytokinesis 1 (PRC1), SMC4 structural maintenance of chromosome 4-like 1 (SMC4L1) and Cyclin B2 (CCNB2) [37–39]. Although these genes are associated with tumour development none have been thoroughly characterized in FAP to date.

Altered Gene Expression in Desmoid Tumours

A limited number of gene expression studies have been performed on desmoid tumours, primarily due to the difficulties in obtaining tissue. Two reports have studied gene expression in desmoid disease using 6.8 K, 19 K and 33 K Affymetrix microarrays [40, 41]. Skubitz and Skubitz (2004) [40] reported that ADAM12, WISP-1, Sox-11 and fibroblast activation protein-a are uniquely expressed in desmoids. Denys et al. (2004) identified 69 differentially expressed genes in desmoid tumour tissue compared to normal fibroblasts, before focusing on the down-regulation of IGFBP-6 [41].

A number of genes that were identified as being differentially expressed in desmoid tumours in this study have been reported previously. The over-expressed genes include transforming growth factor β3 (TGFβ3), a distintegrin and metalloproteinase domain 19 (ADAM19), chimerin 1 (CHN1), and ephrin-B3 (EFNB3) [40, 41]. The under-expressed genes include quiescin Q6 (QSCN6), prostaglandin I2 synthase (PTGIS), proenkephalin (PENK), keratin 18 (KRT18), cytokine receptor-like factor 1 (CRLF1), pentaxin-related gene (PTX3) and endoglin (ENG) [41].

Ephrin-B3, a Wnt Target Overexpressed in Desmoid Tumours

The known Wnt/β-catenin target gene ephrin-B3 [42] has been found in this study to be up-regulated more than 2-fold in desmoid tumours compared to normal fibroblasts. The ephrins are ligands for the EPH receptor family, whose normal function is to organize cell patterning in the intestinal crypts [34]. In addition, more recent observations suggest that ephrins are tumour suppressors, although the mechanism by which this is affected remains to be clarified [3, 43, 44]. Further investigation into the precise role of ephrin-B3 is required before any conclusions can be made regarding its role in desmoid disease.

Wound Healing-Associated Genes Differentially Expressed in Desmoid Tumours

Two genes, transforming growth factor β-3 (TGFβ3) and pleiotrophin (PTN), were found to be differentially expressed in desmoid tumours. Both genes are associated with wound healing and could potentially explain the growth advantage of desmoid tumours [45].

TGFβ3 is a multifunctional protein, having roles in cell proliferation and differentiation during embryogenesis and wound healing [46]. Pleiotrophin has been reported to be strongly expressed in many human cancers, and is thought to promote malignant transformation and angiogenesis [47]. It is also frequently found to be up-regulated during the wound healing process [48].

In this study, three genes associated with negative regulation of the wound response have been identified as being under-expressed in desmoid tumours. The three genes are: signal transducer and activator of transcription 1 (STAT1), mothers against decapentaplegic homolog 3 (MADH3 or Smad3) and mothers against decapentaplegic homolog 6 (MADH6 or Smad6). STAT1 enhances transcription in response to interferon-, an action which has been shown to inhibit the wound healing response by preventing phosphorylation of Smad2 and Smad3 [49]. This in turn inhibits the action of TGFβ on the wound response [50]. The role of Smad3 in the wound response is not entirely understood; however, the absence of Smad3 causes an accelerated healing response, even though its over-expression has also been shown to promote healing [51, 52]. Smad6 is a known inhibitor of TGFβ, and has shown to be down-regulated in keloids [53].

The abundance of wound response-related genes found to be deregulated in the desmoid tumours in this study adds to the notion that desmoid formation is an abnormal wound response. The finding of over-expressed genes involved in fibroblast proliferation and migration could explain the abnormal proliferation and local invasiveness of desmoid tumours. The down-regulation of angiogenesis-associated genes could account for the poor vascularisation of desmoids.

The limiting factor in this study of desmoid tumours is the small number of desmoids available. In order to reach more conclusions regarding the exact molecular nature of desmoids and their growth mechanisms, a much larger sample size would be required.

Comparison of FAP Adenoma and Desmoid Tumour Molecular Profiles

It has long been recognized that desmoid tumours occur with a much higher frequency in FAP patients than in the general population. The apparent role of aberrant Wnt signalling in both diseases could indicate a molecular similarity between the two. Although Wnt target genes were identified as being up-regulated in both tumour types in this study, the specific genes were different in the two groups. The finding of different Wnt targets could be attributed to the use of different control groups for the FAP adenomas and desmoid tumours. Nevertheless, the molecular profiles obtained using cluster analysis clearly demonstrated that FAP adenomas and desmoid tumours display distinctly different gene expression profiles.

References

  1. Ponz de Leon M, Roncucci L: The cause of colorectal cancer. Dig Liver Dis 2000, 32: 426–439. 10.1016/S1590-8658(00)80265-0

    Article  CAS  PubMed  Google Scholar 

  2. Sieber OM, Tomlinson IP, Lamlum H: The adenomatous polyposis coli (APC) tumour suppressor – genetics, function and disease. Mol Med Today 2000, 6: 462–469. 10.1016/S1357-4310(00)01828-1

    Article  CAS  PubMed  Google Scholar 

  3. Giardiello F, Brensinger J, Petersen G: AGA technical review on hereditary colorectal cancer and genetic testing. Gastroenterology 2001, 121: 198–213. 10.1053/gast.2001.25581

    Article  CAS  PubMed  Google Scholar 

  4. Eccles DM, Luijt R, Breukel C, Bullman H, Bunyan D, Fisher A, Barber J, du Boulay C, Primrose J, Burn J, Fodde R: Hereditary desmoid disease due to a frameshift mutation at codon 1924 of the APC gene. Am J Hum Genet 1996, 59: 1193–1201.

    PubMed Central  CAS  PubMed  Google Scholar 

  5. Bonk T, Humeny A, Sutter C, Gebert J, von Knebel Doeberitz M, Becker C-M: Molecular diagnosis of familial adenomatous polyposis (FAP): genotyping of adenomatous polyposis coli (APC) alleles by MALDI-TOF mass spectrometry. Clin Biochem 2002, 35: 87–92. 10.1016/S0009-9120(02)00279-5

    Article  CAS  PubMed  Google Scholar 

  6. Preston SL, Wong WM, Chan AO, Poulsom R, Jeffery R, Goodlad RA, Mandir N, Elia G, Novelli M, Bodmer WF, Tomlinson IP, Wright NA: Bottom-up histogenesis of colorectal adenomas: origin in the monocryptal adenoma and initial expansion by crypt fission. Cancer Res 2003, 63: 3819–3825.

    CAS  PubMed  Google Scholar 

  7. Cruz-Correa M, Giardiello FM: Familial adenomatous polyposis. Gastrointest Endosc 2003, 58: 885–894. 10.1016/S0016-5107(03)02336-8

    Article  PubMed  Google Scholar 

  8. Leslie A, Carey FA, Pratt NR, Steele RJ: The colorectal adenoma–carcinoma sequence. Br J Surg 2002, 89: 845–860. 10.1046/j.1365-2168.2002.02120.x

    Article  CAS  PubMed  Google Scholar 

  9. Scates D, Clark S, Phillips R, Venitt S: Lack of telomerase in desmoids occurring sporadically and in association with familial adenomatous polyposis. Br J Surg 1998, 85: 965–969. 10.1046/j.1365-2168.1998.00720.x

    Article  CAS  PubMed  Google Scholar 

  10. Reitamo J, Scheinin T, Hayry P: The desmoid syndrome. New aspects in the cause, pathogenesis and treatment of the desmoid tumor. Am J Surg 1986, 151: 230–237. 10.1016/0002-9610(86)90076-0

    Article  CAS  PubMed  Google Scholar 

  11. Shields C, Winter D, Kirwan W, Redmond H: Desmoid Tumours. Eur J Surg Oncol 2001, 27: 701–706. 10.1053/ejso.2001.1169

    Article  CAS  PubMed  Google Scholar 

  12. Cheon SS, Cheah AY, Turley S, Nadesan P, Poon R, Clevers H, Alman BA: beta-Catenin stabilization dysregulates mesenchymal cell proliferation, motility, and invasiveness and causes aggressive fibromatosis and hyperplastic cutaneous wounds. Proc Natl Acad Sci USA 2002, 99: 6973–6978. 10.1073/pnas.102657399

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Brueckl WM, Ballhausen WG, Fortsch T, Gunther K, Fiedler W, Gentner B, Croner R, Boxberger F, Kirchner T, Hahn EG, Hohenberger W, Wein A: Genetic testing for germline mutations of the APC gene in patients with apparently sporadic desmoid tumors but a family history of colorectal carcinoma. Dis Colon Rectum 2005, 48: 1275–1281. 10.1007/s10350-004-0949-5

    Article  PubMed  Google Scholar 

  14. Couture J, Mitri A, Lagace R, Smits R, Berk T, Bouchard HL, Fodde R, Alman B, Bapat B: A germline mutation at the extreme 3' end of the APC gene results in a severe desmoid phenotype and is associated with overexpression of beta-catenin in the desmoid tumor. Clin Genet 2000, 57: 205–212. 10.1034/j.1399-0004.2000.570306.x

    Article  CAS  PubMed  Google Scholar 

  15. Alman BA, Li C, Pajerski ME, Diaz-Cano S, Wolfe HJ: Increased beta-catenin protein and somatic APC mutations in sporadic aggressive fibromatoses (desmoid tumors). Am J Pathol 1997, 151: 329–334.

    PubMed Central  CAS  PubMed  Google Scholar 

  16. Cole KA, Krizman DB, Emmert-Buck MR: The genetics of cancer – a 3D model. Nat Genet 1999,21(1 Suppl):38–41. 10.1038/4466

    Article  CAS  PubMed  Google Scholar 

  17. Bucca G, Carruba G, Saetta A, Muti P, Castagnetta L, Smith CP: Gene expression profiling of human cancers. Ann N Y Acad Sci 2004, 1028: 28–37. 10.1196/annals.1322.003

    Article  CAS  PubMed  Google Scholar 

  18. Golub TR, Slonim DK, Tamayo P, Huard C, Gaasenbeek M, Mesirov JP, Coller H, Loh ML, Downing JR, Caligiuri MA, Bloomfield CD, Lander ES: Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. Science 1999, 286: 531–537. 10.1126/science.286.5439.531

    Article  CAS  PubMed  Google Scholar 

  19. Lin Y-M, Furukawa Y, Tsunoda T, Yue C-T, Yang K-C, Nakamura Y: Molecular diagnosis of colorectal tumors by expression profiles of 50 genes expressed differentially in adenomas and carcinomas. Oncogene 2002, 21: 4120–4128. 10.1038/sj.onc.1205518

    Article  CAS  PubMed  Google Scholar 

  20. Azumaya M, Kobayashi M, Ajioka Y, Honma T, Suzuki Y, Takeuchi M, Narisawa R, Asakura H: Size-dependent expression of cyclooxygenase-2 in sporadic colorectal polyps relative to adenomas in patients with familial adenomatous polyposis. Pathol Int 2002, 52: 272–276. 10.1046/j.1440-1827.2002.01350.x

    Article  CAS  PubMed  Google Scholar 

  21. Brabletz T, Herrmann K, Jung A, Faller G, Kirchner T: Expression of nuclear beta-Catenin and c-myc is correlated with tumor size but not with proliferative activity of colorectal adenomas. Am J Pathol 2000, 156: 865–870.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Brosens LA, Iacobuzio-Donahue CA, Keller JJ, Hustinx SR, Carvalho R, Morsink FH, Hylind LM, Offerhaus GJ, Giardiello FM, Goggins M: Increased cyclooxygenase-2 expression in duodenal compared with colonic tissues in familial adenomatous polyposis and relationship to the -765G -> C COX-2 polymorphism. Clin Cancer Res 2005, 11: 4090–4096. 10.1158/1078-0432.CCR-04-2379

    Article  CAS  PubMed  Google Scholar 

  23. El-Bahrawy MA, Talbot IC, Poulsom R, Jeffery R, Alison MR: The expression of E-cadherin and catenins in colorectal tumours from familial adenomatous polyposis patients. J Pathol 2002, 198: 69–76. 10.1002/path.1168

    Article  CAS  PubMed  Google Scholar 

  24. Jungck M, Grunhage F, Spengler U, Dernac A, Mathiak M, Caspari R, Friedl W, Sauerbruch T: E-cadherin expression is homogeneously reduced in adenoma from patients with familial adenomatous polyposis: an immunohistochemical study of E-cadherin, beta-catenin and cyclooxygenase-2 expression. Int J Colorectal Dis 2004, 19: 438–445. 10.1007/s00384-003-0575-z

    Article  CAS  PubMed  Google Scholar 

  25. Khan KN, Masferrer JL, Woerner BM, Soslow R, Koki AT: Enhanced cyclooxygenase-2 expression in sporadic and familial adenomatous polyposis of the human colon. Scand J Gastroenterol 2001, 36: 865–869. 10.1080/003655201750313405

    Article  CAS  PubMed  Google Scholar 

  26. Dang DT, Bachman KE, Mahatan CS, Dang LH, Giardiello FM, Yang VW: Decreased expression of the gut-enriched Kruppel-like factor gene in intestinal adenomas of multiple intestinal neoplasia mice and in colonic adenomas of familial adenomatous polyposis patients. FEBS Lett 2000, 476: 203–207. 10.1016/S0014-5793(00)01727-0

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Salahshor S, Goncalves J, Chetty R, Gallinger S, Woodgett JR: Differential gene expression profile reveals deregulation of pregnancy specific beta1 glycoprotein 9 early during colorectal carcinogenesis. BMC Cancer 2005, 5: 66. 10.1186/1471-2407-5-66

    Article  PubMed Central  PubMed  Google Scholar 

  28. Hlubek F, Pfeiffer S, Budczies J, Spaderna S, Jung A, Kirchner T, Brabletz T: Securin (hPTTG1) expression is regulated by beta–catenin/TCF in human colorectal carcinoma. Br J Cancer 2006, 94: 1672–1677.

    PubMed Central  CAS  PubMed  Google Scholar 

  29. Pfleghaar K, Heubes S, Cox J, Stemmann O, Speicher MR: Securin is not required for chromosomal stability in human cells. PLoS Biol 2005, 3: e416. 10.1371/journal.pbio.0030416

    Article  PubMed Central  PubMed  Google Scholar 

  30. Fujii T, Nomoto S, Koshikawa K, Yatabe Y, Teshigawara O, Mori T, Inoue S, Takeda S, Nakao A: Overexpression of pituitary tumor transforming gene 1 in HCC is associated with angiogenesis and poor prognosis. Hepatology 2006, 43: 1267–1275. 10.1002/hep.21181

    Article  CAS  PubMed  Google Scholar 

  31. Heaney AP, Singson R, McCabe CJ, Nelson V, Nakashima M, Melmed S: Expression of pituitary-tumour transforming gene in colorectal tumours. Lancet 2000, 355: 716–719. 10.1016/S0140-6736(99)10238-1

    Article  CAS  PubMed  Google Scholar 

  32. Cho-Rok J, Yoo J, Jang YJ, Kim S, Chu IS, Yeom YI, Choi JY, Im DS: Adenovirus-mediated transfer of siRNA against PTTG1 inhibits liver cancer cell growth in vitro and in vivo. Hepatology 2006, 43: 1042–1052. 10.1002/hep.21137

    Article  PubMed  Google Scholar 

  33. Drivdahl R, Haugk KH, Sprenger CC, Nelson PS, Tennant MK, Plymate SR: Suppression of growth and tumorigenicity in the prostate tumor cell line M12 by overexpression of the transcription factor SOX9. Oncogene 2004, 23: 4584–4593. 10.1038/sj.onc.1207603

    Article  CAS  PubMed  Google Scholar 

  34. Blache P, Wetering M, Duluc I, Domon C, Berta P, Freund JN, Clevers H, Jay P: SOX9 is an intestine crypt transcription factor, is regulated by the Wnt pathway, and represses the CDX2 and MUC2 genes. J Cell Biol 2004, 166: 37–47. 10.1083/jcb.200311021

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Batlle E, Henderson JT, Beghtel H, Born MM, Sancho E, Huls G, Meeldijk J, Robertson J, Wetering M, Pawson T, Clevers H: Beta-catenin and TCF mediate cell positioning in the intestinal epithelium by controlling the expression of EphB/ephrinB. Cell 2002, 111: 251–263. 10.1016/S0092-8674(02)01015-2

    Article  CAS  PubMed  Google Scholar 

  36. Nakada M, Niska J, Miyamori H, McDonough WS, Wu J, Sato H, Berens ME: The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells. Cancer Res 2004, 64: 3179–3185. 10.1158/0008-5472.CAN-03-3667

    Article  CAS  PubMed  Google Scholar 

  37. Wu Q, Suo Z, Risberg B, Karlsson MG, Villman K, Nesland JM: Expression of Ephb2 and Ephb4 in breast carcinoma. Pathol Oncol Res 2004, 10: 26–33. 10.1007/BF02893405

    Article  CAS  PubMed  Google Scholar 

  38. Jager D, Stockert E, Jager E, Gure AO, Scanlan MJ, Knuth A, Old LJ, Chen YT: Serological cloning of a melanocyte rab guanosine 5'-triphosphate-binding protein and a chromosome condensation protein from a melanoma complementary DNA library. Cancer Res 2000, 60: 3584–3591.

    CAS  PubMed  Google Scholar 

  39. Li C, Lin M, Liu J: Identification of PRC1 as the p53 target gene uncovers a novel function of p53 in the regulation of cytokinesis. Oncogene 2004, 23: 9336–9347. 10.1038/sj.onc.1208114

    Article  CAS  PubMed  Google Scholar 

  40. Sarafan-Vasseur N, Lamy A, Bourguignon J, Le Pessot F, Hieter P, Sesboue R, Bastard C, Frebourg T, Flaman JM: Overexpression of B-type cyclins alters chromosomal segregation. Oncogene 2002, 21: 2051–2057. 10.1038/sj.onc.1205257

    Article  CAS  PubMed  Google Scholar 

  41. Skubitz KM, Skubitz AP: Gene expression in aggressive fibromatosis. J Lab Clin Med 2004, 143: 89–98. 10.1016/j.lab.2003.10.002

    Article  CAS  PubMed  Google Scholar 

  42. Denys H, Jadidizadeh A, Amini Nik S, Van Dam K, Aerts S, Alman BA, Cassiman JJ, Tejpar S: Identification of IGFBP-6 as a significantly downregulated gene by beta-catenin in desmoid tumors. Oncogene 2004, 23: 654–664. 10.1038/sj.onc.1207160

    Article  CAS  PubMed  Google Scholar 

  43. Katoh Y, Katoh M: Comparative integromics on Ephrin family. Oncol Rep 2006, 15: 1391–1395.

    CAS  PubMed  Google Scholar 

  44. Jubb AM, Zhong F, Bheddah S, Grabsch HI, Frantz GD, Mueller W, Kavi V, Quirke P, Polakis P, Koeppen H: EphB2 is a prognostic factor in colorectal cancer. Clin Cancer Res 2005, 11: 5181–5187. 10.1158/1078-0432.CCR-05-0143

    Article  CAS  PubMed  Google Scholar 

  45. Guo DL, Zhang J, Yuen ST, Tsui WY, Chan AS, Ho C, Ji J, Leung SY, Chen X: Reduced expression of EphB2 that parallels invasion and metastasis in colorectal tumours. Carcinogenesis 2006, 27: 454–464. 10.1093/carcin/bgi259

    Article  CAS  PubMed  Google Scholar 

  46. Lee CH, Bang SH, Lee SK, Song KY, Lee IC: Gene expression profiling reveals sequential changes in gastric tubular adenoma and carcinoma in situ. World J Gastroenterol 2005, 11: 1937–1945.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  47. Cox DA: Transforming growth factor-beta 3. Cell Biol Int 1995, 19: 357–371. 10.1006/cbir.1995.1082

    Article  CAS  PubMed  Google Scholar 

  48. Riegel AT, Wellstein A: The potential role of the heparin-binding growth factor pleiotrophin in breast cancer. Breast Cancer Res Treat 1994, 31: 309–314. 10.1007/BF00666163

    Article  CAS  PubMed  Google Scholar 

  49. Christman KL, Fang Q, Kim AJ, Sievers RE, Fok HH, Candia AF, Colley KJ, Herradon G, Ezquerra L, Deuel TF, Lee RJ: Pleiotrophin induces formation of functional neovasculature in vivo. Biochem Biophys Res Commun 2005, 332: 1146–1152. 10.1016/j.bbrc.2005.04.174

    Article  CAS  PubMed  Google Scholar 

  50. DaFonseca CJ, Shu F, Zhang JJ: Identification of two residues in MCM5 critical for the assembly of MCM complexes and Stat1-mediated transcription activation in response to IFN-gamma. Proc Natl Acad Sci USA 2001, 98: 3034–3039. 10.1073/pnas.061487598

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  51. O'Kane S, Ferguson MW: Transforming growth factor beta s and wound healing. Int J Biochem Cell Biol 1997, 29: 63–78. 10.1016/S1357-2725(96)00120-3

    Article  PubMed  Google Scholar 

  52. Falanga V, Schrayer D, Cha J, Butmarc J, Carson P, Roberts AB, Kim SJ: Full-thickness wounding of the mouse tail as a model for delayed wound healing: accelerated wound closure in Smad3 knock-out mice. Wound Repair Regen 2004, 12: 320–326. 10.1111/j.1067-1927.2004.012316.x

    Article  PubMed  Google Scholar 

  53. Sumiyoshi K, Nakao A, Setoguchi Y, Okumura K, Ogawa H: Exogenous Smad3 accelerates wound healing in a rabbit dermal ulcer model. J Invest Dermatol 2004, 123: 229–236. 10.1111/j.0022-202X.2004.22730.x

    Article  CAS  PubMed  Google Scholar 

  54. Yu H, Bock O, Bayat A, Ferguson MW, Mrowietz U: Decreased expression of inhibitory SMAD6 and SMAD7 in keloid scarring. J Plast Reconstr Aesthet Surg 2006, 59: 221–229. 10.1016/j.bjps.2005.06.010

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported in part by funds from the NBN Childhood Cancer Research Group, the University of Newcastle, the Clive and Vera Ramaciotti Centre for Gene Function Analysis, and the Hunter Medical Research Institute (HMRI).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rodney J Scott.

Rights and permissions

Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution 2.0 International License (https://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and permissions

About this article

Cite this article

Bowden, N.A., Croft, A. & Scott, R.J. Gene Expression Profiling in Familial Adenomatous Polyposis Adenomas and Desmoid Disease. Hered Cancer Clin Pract 5, 79 (2007). https://doi.org/10.1186/1897-4287-5-2-79

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/1897-4287-5-2-79

Keywords