69 Id2 expression is inversely correlated with tumour grade in breast cancer JP Bury , SS Cross , S Balasubramanian , MW Reed , CE Lewis University <strong>of</strong> Sheffield, Sheffield, United Kingdom Id proteins are members <strong>of</strong> the basic-Helix-Loop-Helix (bHLH) family <strong>of</strong> transcription factors. <strong>The</strong> four known members <strong>of</strong> the family (Id1, Id2, Id3 and Id4) lack a DNA binding domain, and function as dominant negative regulators <strong>of</strong> DNA transcription by other bHLH transcription factors, with which they heterodimerise and thus inactive. Id2 expression has been shown to be required for the maintenance <strong>of</strong> a differentiated phenotype in breast epithelial cells. In this study we sought to compare levels <strong>of</strong> Id2 expression in breast tumours, as assessed immunohistochemically, with tumour grade. 3 triplicate tissue microarrays were created, each containing 170 1mm diameter cores, one from each <strong>of</strong> 170 breast cancers. <strong>The</strong>se arrays were immunostained using a polyclonal antibody against Id2 (Santa Cruz sc-489). Specificity <strong>of</strong> staining was demonstrated using a specific peptide blocker, which blocked all staining. Each core was assigned a score <strong>of</strong> 0 or 1 on the basis <strong>of</strong> the presence or absence <strong>of</strong> Id2 staining. A composite score was calculated for each tumour by adding together the scores for tissue cores from the same tumour in each <strong>of</strong> the three microarrays. <strong>The</strong> mean total score was 1.81 for grade 1 tumours (n=36), 1.68 for grade 2 tumours (n=79) and 1.05 for grade 3 tumours (n=55). This decrease in Id2 expression with increasing tumour grade was statistically significant (p=0.005, Kruskal Wallis test). To our knowledge, this is the largest series <strong>of</strong> breast tumours in which Id2 expression has been correlated with tumour differentiation, and a clear association between tumour de-differentiation and loss <strong>of</strong> Id2 expression is demonstrated. Studies <strong>of</strong> the prognostic significance <strong>of</strong> Id2 expression in breast carcinoma may be valuable. 70 Should random quadrant samples be taken when assessing a cancer mastectomy specimen? CDT Bratten , P Carder , S Lane , A Hanby Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom When assessing a mastectomy specimen, pathologists take great care to accurately locate and extensively sample all macroscopic or radiologically detected lesions. In addition, it is common practice to randomly sample each <strong>of</strong> the four quadrants. <strong>The</strong> utility <strong>of</strong> so doing has only been studied once (Gupta-Dilip et al., 2003, Breast J., 9(4), 307-11). In this US study <strong>of</strong> 78 mastectomy specimens the authors concluded that random quadrant sampling added clinically useful information in a high proportion <strong>of</strong> cases (27%). <strong>The</strong> relatively high number <strong>of</strong> ‘random’ blocks taken (mean 9, range 2 to 19), and the high percentage <strong>of</strong> cases with positive quadrant findings, lead us to believe that this study might not be representative <strong>of</strong> our experience, where traditionally only two to four random blocks are taken and positive findings were thought to be less frequent. Accordingly, we reviewed all mastectomy specimens over a two year period (2001-2002) received by our hospital. In total 388 reports were examined; in 47 cases (12%), random quadrant blocks were positive for malignant disease (17 in-situ disease, 22 invasive, 8 lymphovascular permeation). In all <strong>of</strong> these cases these features had been noted in the main tumour blocks. <strong>The</strong>re was no association with invasive tumour type and positive quadrant findings. However, there were 14 cases (3%) where the random findings were the only indication <strong>of</strong> disease potentially more widespread than the main tumour blocks (i.e. all nodes negative and lymphovascular invasion not seen). 10 <strong>of</strong> these cases identified further in-situ disease and 4 cases invasive disease in the random quadrants. Nevertheless, the clinical impact these extra findings had on patient management is debatable. 71 Audit Of C3 And C4 Breast Fine Needle Aspirate Cases With Histological Correlation I U Nicklaus-Wollenteit , N Singh , RJ Howitt Southampton University Hospitals NHS Trust, Department <strong>of</strong> Cellular Pathology, Southampton, United Kingdom All fine needle aspirates <strong>of</strong> the breast reported as C3 “atypia probably benign“ or C4 “suspicious <strong>of</strong> malignancy“ during 2002 in a large university teaching hospital department have been audited with regard to frequency and histological outcome. In total 982 cases were reported by one <strong>of</strong> four cytopathologists according to the NHSBSP guidelines for cytology. 74 cases (7.5%) were reported as C3 <strong>of</strong> which 61 proceeded to biopsy, and 42 cases (4.3%) were reported as C4 <strong>of</strong> which 41 had a histological diagnosis. 18 <strong>of</strong> the 61 C3 cases (29.5%) were malignant on subsequent histology: 4 low or intermediate grade DCIS; 11 grade 1 or 2 invasive carcinomas, including lobular, mucinous and adenoid cystic subtypes; and 3 invasive grade 3 carcinomas. Of the 42 C4 cases 31 were confirmed as histologically malignant (73.8%): 25 grade 1 or 2 invasive carcinomas; 4 grade 3 invasive carcinomas; and 2 cases <strong>of</strong> pure DCIS. Fibroadenomas and epithelial hyperplasia accounted for a high proportion <strong>of</strong> benign lesions reported as C4. Comparison with a previous study shows similar results. <strong>The</strong> C3 and C4 categories have distinct histological counterparts, and when used appropriately allow the classification <strong>of</strong> challenging cases into clinically distinct groups with different likelihood <strong>of</strong> malignancy. 72 <strong>The</strong> Retrospective Reconstruction Of Prognosis In Breast Cancer: <strong>The</strong> Use Of <strong>The</strong> Nottingham Prognostic Index In <strong>The</strong> Medico-Legal Arena. C Elston 1 , M Blamey 1 , NA Wright 2 1 Department <strong>of</strong> Histopathology, City Hospital, Nottingham, United Kingdom, 2 Histopathology Unit, Cancer Research (UK), London, United Kingdom INTRODUCTION: <strong>The</strong> Nottingham Prognostic Index (NPI) is well-established in comparative use in clinical trials and for assessing prospective prognosis. Here we assess both its application and success in the retrospective determination <strong>of</strong> prognosis in cases <strong>of</strong> litigation for delayed diagnosis <strong>of</strong> breast cancer. METHODS: the usual request is to reconstruct the prognosis <strong>of</strong> a case <strong>of</strong> breast cancer if diagnosis and treatment had occurred at some time previously, which varies between cases. <strong>The</strong> usually available data are (i) the diameter <strong>of</strong> the tumour at resection; (ii) the grade <strong>of</strong> the tumour at resection and (iii) the nodal status after axillary sampling or clearance. Using published data for the doubling time <strong>of</strong> breast carcinomas, the diameter <strong>of</strong> the tumour at the time in question can usually be calculated within confidence intervals. Published data relating the size <strong>of</strong> the tumour to the probability <strong>of</strong> nodal metastases can be used to assess nodal status at this time for unit grade <strong>of</strong> tumour. <strong>The</strong> NPI can then be calculated for the time(s) <strong>of</strong> alleged misdiagnosis. RESULTS: In many cases, the dataset is sufficient to provide values for the NPI and thus survival probability, and in a considerable number <strong>of</strong> cases the NPI has been accepted by the Court since the index case <strong>of</strong> Judge v Quick, which we believe was the first to show causation in an alleged case <strong>of</strong> missed breast cancer. <strong>The</strong> success <strong>of</strong> this approach can be assessed now from a dataset which includes several hundred cases. Problems which have been encountered include cases where (a) the diameter <strong>of</strong> the diameter at diagnosis/excision cannot be assessed; (b) the nodal status is unknown and (c) where there are metachronous tumours. Areas where disputes in expert evidence arose included disagreements about the growth rate <strong>of</strong> breast carcinoma and the significance <strong>of</strong> the size <strong>of</strong> nodal and systemic metastases. CONCLUSION: the NPI has found an unlooked for niche within the medico-legal field which makes it very valuable so long as the assumptions and constraints implicit in the method are remembered. 44
73 Alterations in SEPT9 Gene Expression in Ovarian Epithelial Neoplasia: an Archival Tissue-Based Analysis M Scott , SEH Russell , PA Hall Queen's University Belfast, Belfast, United Kingdom Several lines <strong>of</strong> evidence indicate that the cytoskeletal septin gene SEPT9 may be involved in human neoplasia as a type II cancer gene where alterations in levels <strong>of</strong> expression are key. SEPT9 has a complex genomic architecture through alternative splicing. Information on the expression <strong>of</strong> SEPT9 in normal and neoplastic tissues is limited. We have therefore developed a semiquantitative reverse transcription-PCR strategy to study SEPT9 expression in formalin-fixed paraffin-embedded tissue. Intron-spanning PCR primers produce small amplicons ranging from 61 to 88 base pairs. Laser microdissection allows identification <strong>of</strong> SEPT9 transcripts from
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