Table 1.1 Mean Standard error P value

Staging

of cancer

T2 233 206 p¼0.86

T3 34.82 25.7

TNM1 29.8 13.8 P¼0.09

TNM4 3.55 2.34

Nodal metastasis N0 54.4 41.5

N2 3.22 2.22 P¼0.33

Recurrence Non recurrence 156 109 P¼0.31

Recurrence 39.9 34.9

Distant metastasis No distant 181 126 P¼0.20

Distant metastasis 16.8 13.0

1140 ABSTRACTS

HSP 70 proteins promotes breast cancer cell survival, yet the role of BAG-

1 in HER2+ cancer is not yet defined.

Methods: Proliferation assays, Western blot analysis, and co-immuno-

precipitation were performed in breast cancer cell lines. A library of com-

pounds predicted to inhibit BAG-1 protein interactions was tested by in-

vitro GST-BAG-1/HSC70 pull-down assay.

Results: HER2-inhibiting drugs caused upregulation of BAG-1L ex-

pression in HER2+ cell lines. An increase in proliferation was observed

in HER2+ cells overexpressing BAG-1L and BAG-1M compared to

BAG-1S or pcDNA controls. siRNA knockdown of BAG-1 showed a de-

crease in proliferation. These effects were abrogated following treatment

with trastuzumab. Thioflavin S inhibition of BAG-1 resulted in synergistic

killing of HER2+ cells treated with trastuzumab. We have developed an

antibody shown to be specific for BAG-1L by Western blot and co-immu-

noprecipitation assays.

Conclusion: Ongoing work involves correlation of our findings with

BAG-1L expression patterns divulged from immunohistochemical studies

of whole sections and tissue microarrays using a previously described

breast cancer cohort. Identification of compounds inhibiting BAG-1/

HSC70 protein interaction in HER2+ cells may lead to the development

of novel anti-cancer therapies.

22. Geminin: A prognostic marker in breast cancer

Sreekumar Sundara Rajan1,2, Andrew Hanby1,2, Kieran Horgan2,

Val Speirs1

1 University of Leeds, Leeds Institute of Molecular Medicine, Leeds, West

Yorkshire, UK2Leeds Teaching Hospitals NHS Trust, Leeds, West Yorkshire, UK

Introduction: Geminin is an endogenous inhibitor of DNA replication

and has been shown to indicate the rate of cell cycle progression in spo-

radic breast cancer. The aim of our study was to evaluate the prognostic

role of geminin expression in breast cancer patients.

Methods: Breast cancer tissue microarrays (TMA) containing 291 tu-

mours were stained using anti-geminin antibody (NCL-L; Mouse mono-

clonal; Novocastra). Slides were scanned using AperioScanScope� and

labelling index [LI] was calculated for geminin expression. ROC was

used to determine the optimum cut-off value for geminin LI (>2%) and

Kaplan-Meier curves were plotted to determine disease free survival

(DFS) and breast cancer specific survival (BCSS).

Results: Our results showed poor BCSS amongst patients with gemi-

nin positivity (56/186) in comparison to those who were negative (12/

105) (Log Rank c2 ¼ 11.89; HR 2.85 (1.53, 5.32); Mean ¼ 146 months,

SD � 68.13). Similarly, DFS was worse in patients with geminin positivity

(Log Rank c2 ¼11.47; p¼0.001; HR¼2.63 (1.47, 4.71)). The tumour size,

nodal status, geminin LI (>2%), grade, mitotic index, ER and LVI were all

significant prognostic factors predicting poor DFS and BCSS. However, on

Cox multivariate regression analysis, only geminin LI (DFS, p¼0.000 (CI

1.67, 6.26); BCSS, p¼0.000 (CI 1.79, 7.53)), nodal status (N3 vs. N0,

DFS, p¼0.000 (CI 2.38, 10.05); BCSS, p¼0.000 (CI 2.88, 12.6)) and tu-

mour size (DFS, p¼.001(1.006, 1.026); BSCC, p¼0.003 (CI 1.006,

1.026)) were found to be independent predictors.

Conclusion: Expression of geminin in breast cancer is a strong and in-

dependent predictor of adverse outcome.

23. MTSS1 expression in human colorectal cancer and its significance

e A cohort study

Afaq Siddiqu, Ye Lin, Rachael Hargest, Mahir Al-Rawi, Weng Jiang1 University Hospital of Wales, Cardiff, UK2Glan Clwyd Hospital, Rhyl, UK

Introduction: Colorectal cancer is the third most common cancer in

the UK. Each year, 38,000 patients are diagnosed with colorectal cancer.

Death among these patients is due to metastasis both locally and system-

ically. Recently, a new molecule has been reported to be a potential metas-

tasis related gene, namely MTSS1. The gene was found to be reduced in

aggressive tumour cell lines. Clinical studies, though limited, have indi-

cated that it may be a metastasis suppressing molecule. This study is to in-

vestigate the MTSS1 expression in normal and cancerous tissue and its

correlation with patient outcome.

Method: The expression of MTSS1 was examined in colorectal cancer

cohort using quantitative gene transcript analysis and immunohistochemi-

cal method.

Results: Analysis of MTSS1 expression levels in IHC colon cancer tu-

mour tissue (n¼94) cohort was significantly reduced compared to non-tu-

mour tissue (n¼80) (normal mean 131.3165+/-SD33.2086 vs. tumour

mean 84.79012+/-SD28.98874 p<0.05). Further MTSS1 transcript levels

were found to be reduced in advance stage of disease, nodal, distant me-

tastasis and local recurrence. (Table1.1) These results are not of statistical

significance because of the small numbers in our cohort. Kaplan eMeier

survival analysis showed patients with higher expression levels of

MTSS1 had longer disease-free survival (p<0.05).

Discussion: In our cohort, low level ofMTSS1 expressionwas found to be

reduced in colorectal tumour samples, in aggressive tumours, nodal metasta-

sis, recurrence and distantmetastasis. The current data suggestMTSS1may be

a putative tumour suppressor gene involved in metastasis of the colon cancer.

24. KRAS and BRAF mutations are rare in anal squamous cell

carcinoma, justifying the use of EGFR targeted therapies

Paul Ziprin, Kathleen Elliott, Temi Lampejo, Ben Poskitt, Robert

Goldin, Susan Cleator, Michael Osborn

Imperial College, London, UK

Introduction: The incidence of anal squamous cell carcinoma (SCC) has

increasedmarkedly and it continues to have a poor prognosis. 36% of patients

with loco-regional disease experience a recurrence,while only 10%of patients

withmetastases survive for 2 years ormore; thus new treatment approaches are

required. Assessment of the frequency of KRAS and BRAF mutation in anal

SCC is essential when assessing the potential therapeutic value of epidermal

growth factor receptor (EGFR) inhibition with monoclonals.

Methods: Patients treated for anal cancer at a regional centre were

identified from an anal cancer database. Analysis of KRAS and BRAF mu-

tation status was performed using pyrosequencing on formalin-fixed, par-

affin-embedded clinical material. Codons 12, 13 and 61 on the KRAS gene

and 599, 600 and 601 on the BRAF gene were analysed.

Results: Twenty-seven samples were analysed. Analysis for KRAS

failed for codon 61 in one sample and for codons 12/13 in a second sam-

ple. In 1/26 (3.8%) a N/c.34G>A KRAS mutation was detected on se-

quencing and a small amount of mutant product detected on

conventional PCR methods confirmed the mutation. 4/27 (15.4%) samples

were found to have BRAF mutations, all V600E.

Conclusion: KRAS and BRAF mutations are uncommon in anal SCC,

justifying the potential use of anti-EGFR receptor therapy. Such a treatment

approach requires assessment in clinical trials. KRAS assessment is not

routinely undertaken prior to treatment of head and neck SSC with anti-

EGFR monoclonal antibodies and we have not demonstrated a definite

need in the context of anal SCC.