P57Meta-clustering <strong>of</strong> Small Intestinal, Gastric and ColorectalAdenocarcinomas Based on DNA Copy Number Pr<strong>of</strong>iles.JC Haan 1 ,TE Buffart 1 , MA Van De Wiel 1 , B Carvalho 1 ,CPostma 1 , NCT Van Grieken 1 , ID Nagtegaal 2 ,CJ Mulder 1 ,K Maude 3 , P Howdle 3 ,PQuirke 3 , H Grabsch 3 ,BYlstra 1 ,GA Meijer 11 VU University Medical Centre, Amsterdam, <strong>The</strong> Netherlands, 2 UniversityMedical Centre St. Radboud, Nijmegen, <strong>The</strong> Netherlands, 3 St James’sUniversity Hospital, <strong>Leeds</strong>, UKAdenocarcinomas <strong>of</strong> the stomach, small intestine and colorectum share severalclinical and phenotypical characteristics. In addition, the majority <strong>of</strong> theseadenocarcinomas show chromosomal instability, which results in DNA copynumber aberrations that can be measured by array comparative genomichybridization (aCGH). To which extent these three tumour types share DNAcopy number pr<strong>of</strong>iles is unknown. We and others evaluated the chromosomalgains and losses by array CGH <strong>of</strong> colorectal and gastric tumours. Smallintestinal cancer is rare in comparison to colorectal and gastric cancer and highresolution aCGH data were not previously reported to our knowledge.Previously, our group succeeded to classify 373 epithelial tumours according totheir organ <strong>of</strong> origin based on their aCGH pr<strong>of</strong>iles by hierarchical clustering(Jong et al., Oncogene 2007). Within this meta-cluster <strong>of</strong> primarily epithelialtumours, the gastrointestinal pr<strong>of</strong>iles split into a cluster with primarily colontumours and one cluster with both gastric and colon tumours. No smallintestinal tumours were included.<strong>The</strong> aim <strong>of</strong> the present study is to investigate DNA copy number pr<strong>of</strong>iles <strong>of</strong>small intestinal tumours in comparison to gastric and colorectal.Materials: aCGH data from 35 adenocarcinomas <strong>of</strong> each organ were selectedfrom different in house datasets, based on array quality and clinical data.Smoothed data were called by the calling algorithm CGHcall. Called data wereanalyzed with hierarchical clustering and with frequencies <strong>of</strong> aberrations perorganResults: First results indicate substantial overlap <strong>of</strong> DNA copy number pr<strong>of</strong>ilesbetween small intestinal and colorectal cancer and less overlap with gastriccancer.P59Fragile Histidine Triad (Fhit) and Ki67 Expression inAberrant Crypts and Colorectal Carcinomas.KVaiphei 1 ,ARanga 1 ,JDev Wig 1 , K Joshi 11 Post Graduate Institute <strong>of</strong> Medical Education and Research, Chandigarh,India.Background: Abnormal Expression <strong>of</strong> Fhit Protein is Observed in DifferentTypes <strong>of</strong> Human Malignancies. <strong>The</strong>re Are Limited Literature Analysing Fhitand Ki67 Co-expression in Colorectal Lesions. Objective: ComparativeAnalysis <strong>of</strong> Fhit and Ki67 Expressions by Colorectal Aberrant Crypts (ACF)and Adeno Carcinoma (Adeno Ca). by Immunohistochemistry. Methods:Ninety-five Colonic Adeno Ca were Examined to Identify ACF in MucosaAdjacent to Tumour. Sections Bearing ACF and Tumour were Used for theStudy. Immunohistochemistry for Fhit and Ki67.Results: All had Hyperplastic ACF (HACF) in Adjoining Mucosa, 31% hadDysplastic ACF (DACF). Normal Crypt Epithelium Showed StrongCytoplasmic Expression for Fhit, Stronger along Surface Epithelium, SimilarPattern Observed in HACF, with Stronger Expression by Surface Epithelium.28.57% <strong>of</strong> DACF were Negative and 71.43% Showed Weak to ModerateIntensity. Six <strong>of</strong> Adeno Ca Showed Faint Cytoplasmic Positivity. HACF,DACF and Adeno Ca had Significantly Different Fhit Expression (p65 years (60%), colonic tumours (61%)and Dukes A/B disease (59%). Characteristics were similarto Klintrup's population (DukesA/Bs 61%). Univariate analysis: age (p
P61Proportion <strong>of</strong> Tumour Cells and Stroma – An Inexpensive butReliable Predictor <strong>of</strong> Patient Survival in Colorectal CancerM Dattani 1 , P McShane 2 , D Treanor 1 , G Hutchins 1 , J Grabsch 1 ,JM Brown 2 , H Thorpe 2 , DG Jayne 3 , PJ Guillou 3 ,WMueller 4 ,PQuirke 1 , H Grabsch 11 Pathology and Tumour Biology, <strong>Leeds</strong> Institute <strong>of</strong> Molecular Medicine,University <strong>of</strong> <strong>Leeds</strong>, UK, 2 Clinical Trials Research Unit, University <strong>of</strong><strong>Leeds</strong>, UK, 3 Academic Unit <strong>of</strong> Surgery, St James's University Hospital,<strong>Leeds</strong> Teaching Hospitals NHS Trust, UK, 4 GemeinschaftspraxisPathologie, Starnberg, GermanyRecent studies indicate that tumour growth is determined by cancer cellsthemselves as well as by tumour stroma. Although expression studies haveshown that stroma-like gene expression patterns are related to cancerprogression, tumour components have not been quantified in a large series <strong>of</strong>colorectal cancer (CRC).Performing point counting on virtual HE stained slides, we analysed therelative proportion <strong>of</strong> tumour and stroma in two independent CRC series: CRC-G (n=147, consecutive cases from one German hospital) and CLASICC(n=150, cases from a UK randomised clinical trial). Results were comparedwith clinicopathological parameters and patient survival. Cut <strong>of</strong>fs for Kaplan-Meier analyses were established using ROC curve analyses.High tumour cellularity was associated with prolonged survival in CRC-G(p=0.011) and in CLASICC (p=0.033) and high proportion <strong>of</strong> stroma wasrelated to poor patient survival in CRC-G (p=0.002) and in CLASICC(p=0.015). No significant associations were found with any otherclinicopathological parameters.This is the first study that objectively quantified the morphological tumourcomponents in a large series <strong>of</strong> CRC. It demonstrates that this inexpensivemorphometric method can reproducibly predict patient prognosis in CRCconfirming data from a previous much smaller study in CRC. Although, wecurrently do not fully understand the molecular mechanisms that regulate theproportion <strong>of</strong> stroma in a tumour, the morphological quantification <strong>of</strong> tumourcomponents may be a useful marker for patient stratification to therapiestargeting tumour stroma .P63Routine Staining <strong>of</strong> Gastrointestinal Biopsies in the UnitedKingdomMKoenig 1 , JB Sch<strong>of</strong>ield 21 Brighton & Sussex University Hospitals NHS Trust, 2 Maidstone &Tunbridge Wells NHS TrustA survey was conducted into the routine staining <strong>of</strong> gastrointestinal biopsies inhistopathology departments within the NHS. We aimed to compare the sole use<strong>of</strong> HE staining with the use <strong>of</strong> HE combined with special stains according to thebiopsy site. This project has been endorsed by the Pathology Section <strong>of</strong> theBritish <strong>Society</strong> <strong>of</strong> Gastroenterologists.167 histopathology departments in the United Kingdom were contacted usingan email questionnaire. <strong>The</strong> following question was asked: Which stains areroutinely used in your department when staining biopsies from the oesophagus,stomach, duodenum, small bowel and large bowel?Valid return rate was 92 <strong>of</strong> 167 departments (55%). Approximately two thirds<strong>of</strong> departments only employ HE for assessment <strong>of</strong> oesophageal biopsies. <strong>The</strong>distribution with regard to staining <strong>of</strong> gastric biopsies was approximately equalwith 50% HE-only to 50% HE combined with special stains (mostly AB-PASand Giemsa). Duodenal, small and large bowel biopsies are mostly stained withHE-only. No significant difference could be identified between the use <strong>of</strong>special stains in district general hospitals as opposed to university, teaching orexpert centres.<strong>The</strong> results <strong>of</strong> the survey show routine use <strong>of</strong> special stains to be variable,especially for oesophageal and gastric biopsies. A review <strong>of</strong> the literature hasshown special stains to enhance sensitivity and specificity for the detection <strong>of</strong>pathological abnormalities, especially metaplasia and infections. <strong>The</strong> diversity<strong>of</strong> routine staining practice within the NHS highlights the need to discussguidelines. A consensus on best practice is desirable.P62A Novel SNP in the 5'UTR <strong>of</strong> ATP5A1 is Associated withReduced Gene Expression in Colorectal Cancer Cell LinesJ Keeley 1 ,RSeth 1 ,SCrook 1 , D Jackson 1 , MIlyas 11 Department <strong>of</strong> Pathology, QMC, NUH. NottinghamIntroductionAllelic imbalance (AI) at 18q is a common event in colorectal cancer (CRC).Recently, a loss <strong>of</strong> function mutation in the gene Atp5a1 has been found to beassociated with increased risk <strong>of</strong> tumour progression in Min (Multiple IntestinalNeoplasia) mice. <strong>The</strong> human ortholog maps to chromosome 18q and thus weinvestigated whether ATP5A1 could be a target <strong>of</strong> 18q allelic loss.MethodsEighteen CRC cell lines were studied for ATP5A1 mutation using PCR andHigh resolution Melting analysis. Samples showing aberrant melting curves onthe HRM were sequenced. <strong>The</strong> expression <strong>of</strong> ATP5A1 mRNA was investigatedusing real-time quantitative PCR.ResultsHRM analysis identified several previously described SNPs but loss <strong>of</strong>function somatic mutations were not found within ATP5A1. A novel A to Gtransition was discovered in exon 1. This change is located in the 5’UTR <strong>of</strong>ATP5A1 and is present in LOVO/HCT116 (heterozygous) andRKO/SW480/SW620 (homozygous). All cell lines expressed ATP5A1 mRNAand further analysis demonstrated that cell lines carrying this change expressedlower levels <strong>of</strong> ATP5A1 mRNA than those with the wild type sequence(p = 0.05).ConclusionSomatic mutations <strong>of</strong> ATP5A1 were not found in this study. <strong>The</strong> identicalnature <strong>of</strong> the sequence change in the 5’UTR in 5 different cell lines suggest thatthis is more likely a novel SNP than a somatic change. <strong>The</strong> lower level <strong>of</strong> geneexpression associated with this change is <strong>of</strong> uncertain significance but may givean increased risk <strong>of</strong> tumour progression.P64Perforated Amoebic Colitis Complicating Cortico-steroid andChemotherapy for Metastatic Adenocarcinoma <strong>of</strong> UnknownOriginN Momtahan 1 ,DScott 2 , JP Narain 21 Bradford Royal Infirmary, 2 Harrogate District HospitalFulminant amoebic colitis is a rare form <strong>of</strong> amoebiasis in which thetrophozoites <strong>of</strong> Entamoeba histolytica invade the colonic wall resulting inmassive necrosis <strong>of</strong> wide segments <strong>of</strong> large bowel. It begins as a mucosal ulcer,which expands within the submucosa.Fulminant amoebic colitis is associated with a high mortality despiteaggressive treatment with anti- microbial agents. It has been reported in patientswith asymptomatic amoebiasis who are given corticosteroid for anotherindication and should be considered in the differential diagnosis <strong>of</strong> patients whodevelop fever and diarrhoea after starting such therapy. In fact, corticosteroiduse is the one factor that predicted the likelihood <strong>of</strong> multi-organ failure inpatients with fulminant amoebic colitis. Despite a high incidence <strong>of</strong> colonicperforation (some series reporting incidences as high as 87%, the majority <strong>of</strong>these having multiple perforations), the finding <strong>of</strong> a s<strong>of</strong>t compressible abdomenis present in as many as 65% <strong>of</strong> cases <strong>of</strong> fulminant amoebic colitis. Earlylaparatomy may decrease mortality from nearly 100% to 40%. Demonstration<strong>of</strong> erythrophagocytic amoebae in the stool by microscopic examination is 100%specific for invasive amoebiasisWe hereby present a case that developed fulminant amoebic colitis andcolonic perforation after receiving corticosteroid and chemotherapy formetastatic adenocarcinoma <strong>of</strong> unknown origin.<strong>Summer</strong> <strong>Meeting</strong> (194 th ) 1–4 July <strong>2008</strong> Scientific Programme47
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