bluuebunny/crossref_metadata_2025 huggingface dataset:
P53 functional assay in yeast: evaluation in 1856 patients in a large prospective clinical trial (EORTC 10994/BIG 00-01).
Authors: Bonnefoi H , Zimmer AS , Piccart M , Mauriac L , Campone M , Tubiana-Hulin M , Petit T , Rouanet P , Welnicka-Jaskiewicz M , Cufer T , Blot E , Becette V , Bogaerts J , André S , Cameron D , Bergh J , Iggo R
Published: December 2010

Abstract #1067

Background:Although mutations in the p53 gene can occur in 20-30% of sporadic breast cancer, and are associated with other poor prognostic factors, there is no clear evidence as to whether p53 status should influence therapy. Immunohistochemistry has been demonstrated to be an inferior method compared with sequencing, but the optimal technique to assess the functional p53 status is not known. The yeast test is a functional assay which tests the transcriptional competence of p53, and detects biologically important mutations. Here we present the results of p53 evaluation using this method in a prospective study.  Material and methods: EORTC 10994/BIG 00-01 trial is a randomized trial comparing the neoadjuvant use of an anthracycline-based regimen with a taxane-containing regimen in patients with large operable or locally advanced breast cancers. Fresh frozen tumour biopsies were mandatory before starting chemotherapy. Frozen sections were examined centrally and the p53 test was done if the invasive tumour cell content was above 20%. RNA extract was used to assess p53 status: p53 wild type tumours were defined as tumours whose cDNA transfected in yeast gave less than 20% red colonies (background); p53 mutated tumours gave 20% or more red colonies.  Results: The trial accrual was completed in November 2006 after inclusion of 1856 patients. There was no frozen sample available in 70 patients. In 276 patients the % of tumour cells was <20. The p53 test failed in 62 patients, generally because the quality or quantity of RNA was too low for RT-PCR. P53 was successfully assessed in 1458 patients: 661 (45%) tumours are p53 mutated and 797 (55%) are p53 wt. p53 mutated and wild type tumours were grade III in 40% (265/661) and 19% (153/797), respectively, and estrogen receptor negative in 41% (270/661) and 14.5% (116/797), respectively. We did not see any correlation between p53 status and tumour size or nodal status at diagnosis. HER2 status data are being collected.  Conclusion: P53 status has been successfully evaluated using a yeast assay in 78.5% (1458/1856) of cases in the context of a multicenter trial. In indirect comparisons the frequency of patients with functionally altered p53 status is higher than with sequenced based methods. Our results confirm an association between p53 mutation and high grade or estrogen receptor negative status, but not with age, tumour size or clinical nodal status. More data regarding the quality and quantity of RNA collected will be presented at the meeting.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1067.
Stem cell and epithelial-mesenchymal transition markers on circulating tumor cells in patients with metastatic breast cancer.
Authors: Aktas B , Tewes M , Hauch S , Fehm T , Kimmig R , Kasimir-Bauer S
Published: December 2010

Abstract #107

Background: Stem cell like tumor cells have been suggested to be the active source of metastatic spread in primary tumors. Furthermore, these cells may undergo phenotypic changes, known as epithelial-mesenchymal transition (EMT), which allows them to travel to the site of metastasis formation without getting affected by conventional treatment. Assuming that metastasis requires a dissemination of tumor stem cells or tumor cells showing EMT, it was the purpose of this study to identify these cells among circulating tumor cells (CTC) in blood samples of 28 metastatic breast cancer patients.  Materials and Methods: 5 ml blood was analyzed for EpCAM, MUC-1 and HER-2 transcripts with the AdnaTest BreastCancer (AdnaGen AG). The recovered c-DNA was additionally multiplex tested for three EMT markers [Twist, Akt2, PI3K (TAP)] and separately for the tumor stem-cell marker ALDH1. The analytical sensitivity was determined by the detection of a low number of target cells (5 IGROV cells spiked into 5 ml blood of healthy donors) using the AdnaTest BreastCancer procedure. The identification of EMT markers was considered positive if at least one marker was detected in the sample. Healthy donor samples without spiked tumor cells were used to determine the specificity of the test. The resulting PCR fragments were separated and quantified using the Agilent Bioanalyzer 2100.  Results: Applying an amplicon cut-off value of 0.2 ng/μl for Akt2, 0.15 ng/μl for Twist, 0.25 ng/μl for PI3K and 0.15 ng/μl for ALDH1, 97% of 30 healthy donor samples investigated were negative for TAP and 95% for ALDH1 transcripts. The spiking experiments revealed 80% recovery of the IGROV cells. CTC were detected in 12/28 (43%) breast cancer samples. All samples were further examined for tumor stem cell or TAP markers. In the CTC (+) group 50% were positive for at least one of the TAP markers and 42% for ALDH1. In the CTC (-) group the percentages were 19% and 12%, respectively. The expression of TAP and/or stem cell markers in CTC was compared with the clinical follow up results. In CTC (+) patients diagnosed as non-responders the expression of TAP and ALDH1 was found in 5/12 (40%) samples. In the CTC (+) responder group only one sample expressed TAP and no positive case was found for ALDH1. In the CTC (-) non-responder group 1/14 samples was positive for TAP and ALDH1. 2/14 (14%) of CTC (-) responders were positive for TAP and 1/14 for ALDH1, respectively.  Conclusion: A major proportion of CTC found in the blood of metastatic breast cancer patients shows EMT and tumor stem cell characteristics supporting the hypothesis that these markers are an indicator for therapy resistant tumor cell populations and, therefore, for an inferior prognosis.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 107.
Molecular breast cancer prognosis in node negative early breast cancer.
Authors: Schmidt M , Tramm T , Böhm D , Kyndi M , Lehr H , Alstner J , Kölbl H , Stropp U , von Törne C , Weber K , Hennig G , Gehrmann M , Overgaard J
Published: December 2010

Abstract #1071

Background: A number of molecular signatures have been published to aid breast cancer prognosis and therapy response prediction. However, most signatures rely on measurements of dozens of genes on an array platform. Here we report the identification and validation of a prognostic algorithm based on a small number of expressed genes reflecting dominant prognostic motifs like steroid hormone receptor expression, proliferation, matrix degradation and B-cell infiltration. The test is designed for mRNA extracted from formalin fixed paraffin embedded tumour tissue (FFPE).  Materials and Methods: 200 fresh frozen tumours from node negative patients, who did not receive systemic therapy after surgery, were profiled on HG-U133a arrays. Primer and probes were designed for genes identified as prognostic relevant and used to profile the corresponding paired FFPE tumour tissue of our identification cohort by kinetic PCR. A genomic algorithm was constructed and tested in FFPE tumor tissue of an independent validation cohort of 240 node negative patients that remained untreated in the adjuvant setting.  Results: Based on the Prognostic-score of our algorithm we classified the patients of the validation cohort into three classes: low risk (25% of patients), intermediate risk (25%) and high risk (50%). The probability to die after having a recurrence within 10 years was 4.7% for the low risk group, 11.9% for the intermediate risk group and 33.1% for the high risk group. Kaplan Meier analysis revealed that the trend for survival (Logrank test for trend P = 0.0002) was highly significant. Adjuvant! Online calculated probabilities to die of cancer were also used to classify patients into three risk groups of comparable size (25/25/50 percent of patients). The observed risk was 9.1% in the low risk, 26.7% in the intermediate and 23.6% in the high risk group. The area under the ROC curve for Adjuvant! Online was 0.619 (95% CI, 0.52 to 0.718; P = 0.031) while for our algorithm it was 0.755 (95% CI, 0.6628 to 0.8473; P < 0.0001).  Discussion: We identified a prognostic algorithm for node negative breast cancer based on kinetic PCR analysis of FFPE tumor tissue. Validation in an independent cohort showed the ability of the algorithm to classify patients into clinically relevant risk classes. Preliminary analysis shows a better prognostic performance than current clinical guidelines.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1071.
Significantly higher expression of vascular endothelial growth factor (VEGF) and shorter survival after recurrences in premenopausal node negative patients with triple negative breast cancer.
Authors: Linderholm BK , Klintman M , Grabau D , Bendahl P , Fernö M , Per M
Published: December 2010

Abstract #1077

Background: Triple-negative breast cancer (TNBC) amounts to approximately 15% of all breast cancer and is defined as lacking expression of oestrogen- and progesterone receptors (ER and PgR) and HER2 leaving chemotherapy as the only treatment option. The aggressive nature of TNBC is reflected by high histological grade, early onset of relapses, and more commonly visceral metastases. Further exploration of biological features in this group could (would) help to find potential targeted therapies. Aim: To compare intratumoural levels of VEGF and survival in patients with TNBC compared with HER2 positive patients and ER and/or PgR positive but HER2 negative patients. Patients and methods: 210 premenopausal patients with lymph-node negative breast cancer included in a prospective clinical trial were studied. (Ref Malmström et al JCO 19:2010-2019, 2001). Levels of VEGF were determined by an enzyme-linked immuno-sorbent assay (ELISA) on lysates from freshly frozen breast tumor material. HER2 amplification was determined by in situ hybridization of tumor with Her2 + or 3+ reactivity with IHC (determined by IHC followed by confirmatory FISH in 2+ or 3+). Results: Thirty-six out of 210 (17%) patients were classified as having a TNBC. The median level of VEGF in the total patient population was 81 ng/mg total protein (range 2.0- 3610). Significantly higher VEGF levels were found in TNBC compared with HER2 pos. or HER2 neg. ER/PgR pos. patients; median levels 250, 157 and 53 ng/mg total protein respectively (p<0.001). Similarly, 84% of TNBC had VEGF above the median compared to 62% in HER2 pos and 41% in the HER2 neg. ER/PgR pos. group (p<0.001). TNBC was significantly associated to high histological grade (89% grade III) (p<0.001), and high S-phase fraction (p=0.001). The highest rate of distant metastases was (seen or) recorded for HER2 pos patients while TNBC and HER2 neg. ER/PgR pos. had similar relapse rates (p<0.001; three-group comparison). After adjustment for chemotherapy, patients with TNBC survived significantly shorter after relapse compared with ER/PgR pos/HER2 neg patients (HR=7.2;95%CI=2.1-25; p=0.002) but not compared to HER2 pos patients (HR=2.3;95%CI=0.73-7.0; p=0.16). Conclusion: We here for the first time show significant higher levels of an angiogenesis stimulating factor, VEGF in TNBC. Whether blockade of angiogenesis can be an effective new targeted treatment option for this poor prognosis group will be elucidated in ongoing trials.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1077.
Breast cancer molecular subtypes and the risk of second malignancies.
Authors: Radecka B , Hudala-Klecha J , Badzio A , Szwiec M , Drosik K , Jassem J
Published: December 2010

Abstract #1078

Background: Particular molecular subtypes of breast cancer (BC) have been shown to manifest various clinical behaviors. In a large consecutive series of pts we compared treatment outcomes of four major BC subtypes, with particular focus on the risk of second malignancies.  Materials and methods: Triple negative breast cancer (TNBC) was characterized by negativity of hormone receptors (HR) and HER2 (0-2+ by immunohistochemistry). Other subtypes included HR+/HER2-, HR+ /HER2+ and HR-/HER2+. All pts underwent curative surgery from January 2003 to December 2007, followed by radiation and/or systemic therapy according to current standards. Univariate risks of relapse and death were estimated using Kaplan-Meier survival curves and compared with log-rank test, and the multivariate analysis used stratified Cox model.  Results: Of the 783 tumors, 520, 50, 48 and 165 were classified as HR+/HER2-, HR+ /HER2+, HR-/HER2+ and TNBC, respectively. The TNBC and HR-/HER2+ subtypes were associated with a higher tumor grade (p<0.001), higher T stage (p<0.001) and higher proportion of medullary type (p<0.001) compared to other two subgroups. Other characteristics, such as age and menopausal status showed no difference. The probability of relapse-free survival (RFS) and overall survival (OS) in TNBC and HR-/HER2+ groups were similar, and lower than in other two types (TNBC vs. HR+/HER2- and HR+ /HER2+; p=0.002 and 0.03 for RFS and OS). Impaired prognosis of TNBC and HR-/HER2+ pts remained significant after correction by T and N stage. TNBC was associated with almost doubled risk of second malignancy (12.7 vs. 6.5%) compared to all other types (p=0.008). This risk was significantly increased for contralateral BC and endometrial cancer (p=0.002 and 0.024, respectively).  Conclusion: TNBC and HR-/HER2+ subtypes are associated with worse prognosis. Additionally, TNBC carries an increased risk of second malignancies, most likely due to higher proportion of hereditary BC.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1078.
Improving NPI for breast cancer prognosis by including PR and HER-2 expression: own data and external validation set.
Authors: Van Belle V , Brouckaert O , Van Huffel S , Schlichting E , Synnestvedt M , Naume B , Christiaens M , Neven P
Published: December 2010

Abstract #1080

The Nottingham Prognostic Index (NPI) classifies primary operable breast cancer patients into good (NPI ≤ 3.4) , intermediate (NPI 3.4-5.4) and poor (NPI ≥ 5.4) prognostic risk groups [Br J Cancer 1987; 56: 489–92]. Studies indicate a significant short term prognostic role for estrogen (ER), progesterone (PR) and HER-2 receptor on disease free survival (DFS). We proposed at SABCS 2007 to add PR and HER-2 expression to the NPI called improved NPI (iNPI) defined as (NPI) + (HER-2) – (PR), with HER-2 and PR '1' in case of amplified HER-2 and any PR-expression and '0' otherwise.  Internal validation of the iNPI was performed on 1928 breast cancer patients, primary operated between Jan 2000 and June 2005 at Univ Hospital Leuven. All patients were classified into any of the three NPI or iNPI risk group. Mean follow-up was 3.5 yrs and DFS was defined as previously stated [Breast Cancer Research and Treatment 2007; 106: S247-S247]. Out of the 601 patients classified as NPI low risk, 59 shifted towards the intermediate iNPI group. Comparison of DFS for the 59 shifted with the non-shifted patients from the low NPI class did not reveal a significant difference (p=0.129, log-rank test). 57 patients out of 935 and 136 out of 392 respectively classified as intermediate and high NPI group became high and intermediate risk according to the iNPI. In both cases, DFS of the shifted patients differed significantly from DFS of the non-shifted patients (p=<.001 en p=0.029). DFS of the shifted patients with the iNPI group they where shifted to was not/marginally significant (p=0.856, p=0.462 and p=0.047 for low, intermediate and high NPI respectively).  For external validation a dataset from the Breast Cancer Micrometastasis group in Oslo with 677 patients and known NPI, PR and HER-2 was used. Figure 1 confirms a significant difference between DFS of the shifted and the non-shifted patients within the highest NPI risk group (p=0.021). The difference in DFS between shifted and non-shifted patients was marginally non-significant (p=0.068) in the intermediate and non-significant in the low NPI group (p=0.116). The non-significant difference between DFS of the shifted groups with the DFS of the group they were shifted to was confirmed, with exception of the low NPI group (difference in systemic treatment) (p=0.040, p=0.621 and p=0.076 for low, intermediate and high NPI respectively).  We confirm the results from an internal study at the Univ Hospital in Leuven on an external dataset. Inclusion of the PR and HER-2 status into the NPI results in more homogenate risk groups concerning DFS.   

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1080.
Benefit of the 70-gene profile for widely used guidelines: an answer to increased selection for adjuvant chemotherapy in breast cancer.
Authors: Knauer M , Retèl V , Bueno-de-Mesquita J , Rutgers E , van Harten W , Rodenhuis S , van de Vijver M , van 't Veer L , Linn S
Published: December 2010

Abstract #1084

Introduction:  Risk assessment for breast cancer patients differs substantially among treatment guidelines. The NCCN, St.Gallen, Adjuvant!Online, and Dutch 2008 guidelines are less restrictive in comparison to the 2004 Dutch guidelines and Nottingham Prognostic Index, when selecting patients for adjuvant systemic treatment. The Dutch Institute for Healthcare Improvement (CBO) has introduced slight changes in the concept 2008 guidelines. Adjuvant systemic treatment is only advised when the absolute 10-years survival benefit is 5% or more. The new recommendations for adjuvant systemic treatment are based on survival tables used in the Adjuvant! software. The changes affect only node-negative patients aged 35 years: since 2008 patients with G2/G3 tumors >1cm or every tumor >2cm are advised to undergo adjuvant systemic therapy, while in 2004 this was the case for tumors G3>1cm, G2>2cm or every tumor >3cm.  Patients and Methods:  Risk was assessed for 427 lymph-node negative (LNN) patients in the prospective RASTER-study (Bueno-de-Mesquita, 2007) and 151 LNN patients in the NEJM-series (van de Vijver, 2002). Clinical risk was calculated using the 2004 and 2008 Dutch guidelines. Genetic risk was assigned according to the result of the Amsterdam 70-gene signature. Survival analyses were done according to the univariate Kaplan-Meier-method.  Results:  Instead of 57% in 2004, in 2008 only 24% of patients were assigned to clinical low risk in the RASTER study, and 141 patients (33%) changed from low to high risk (p<0.001). The rate of discordant findings between clinical assessment and 70-gene signature increased from 30% in 2004 to 41% in 2008 (p<0.001, table 1). Similar results were obtained for the 151 patients of the NEJM-series (30% and 35% discordance for the 2004 and 2008 CBO guidelines, respectively). At 10 years follow-up, differences between the high and low risk categories according to the 2004, 2008 guidelines and the 70-gene profile, were best predicted by the latter: distant-disease-free survival (DDFS) log-rank p=0.002, p=0.11 and p<0.001 respectively.    Conclusion:  As adjuvant systemic treatments become more effective, guidelines become less restrictive, resulting in more patients being selected for adjuvant therapy. Since newer treatment guidelines do not better predict survival, the magnitude of the 70-gene profiles benefit, which is defined as proportion of patients in whom over- and undertreatment can be avoided, depends on present guidelines for risk assessment and shows a remarkable increase.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1084.
Metaplastic breast carcinoma: clinicopathologic features and prognostic value of triple-negativity.
Authors: Lim K , Oh D , Chie E , Han W , Im S , Kim T , Park I , Nho D , Ha S , Bang Y
Published: December 2010

Abstract #1085

Backgroud Metaplastic breast carcinomas (MBC) are a rare type of breast cancer comprising <1% of all invasive breast cancers and are generally characterized by hormone receptor and human epidermal growth factor receptor 2 (HER2) negativity. There is a paucity of information on prognosis according to hormone receptor and HER2 expression for these rare tumors. The aim of this study was to compare the clinical features and prognosis, between triple negative metaplastic carcinoma (TNMC) and non-triple negative metaplastic carcinoma (NTNMC).  Material and Methods We retrospectively analyzed MBC patients treated at Seoul National University Hospital between 1996 and 2006. Pathologic, immunohistochemical findings and clinical outcome information were reviewed.  Results fifty-one patients were identified. The median age at presentation was 45.8 years (range: 27.3-83.8). Median tumor size at diagnosis was 3.0 cm (range: 0.8-12.0). 34 cases (66.7%) were node-negative, 16 (31.4%) node-positive, and 1 (2.0%) were missing. Estrogen receptor (ER)/progesterone receptor (PR) yielded negative results in 49 cases (96.1%) and 48 cases (94.1%), respectively. HER2 overexpression by immunohistochemistry was negative in 41 of 51 (80.4%). At median follow-up of 40.8 months, the 5-year overall survival rate and disease free survival were 73.9% and 64.9%, respectively. Non-triple negativity (p=0.031) correlated significant with overall survival in multivariate analysis.  Of the 51 patients, 39 (76.5%) were TNMC, and 12 (23.5%) were NTNMC. In TNMC and NTNMC group, median ages were 45.6 and 51.5 years and tumor sizes were 3.0 cm and 3.0 cm, respectively. There were 12 patients (30.8%) in TNMC and 4 patients (33.3%) in NTNMC for lymph node metastasis. Two groups did not differ significantly by age, tumor size, or nodal status. Positive rates for ER, PR and HER2 were 16.7%, 25% and 83.3% in NTNMC. The 3-year overall survival rates in TNMC and NTNMC were 91.4% and 61.9%, respectively (p=0.029). As regards to 3-years disease-free survival, there was no statistically significant difference between TNMC and NTNMC (p=0.694, 75.8% versus 75.0%).  Discussion MBC mainly has triple negative features. However, in subgroup analysis of MBC, non-triple negative group displayed a poor prognosis compared with triple negative group, which is contrary to the case of invasive ductal carcinoma of breast. Further research exploring mechanism of this result.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1085.
Ki-67 gives additional prognostic information on St Gallen 2007 and Adjuvant! Online risk categories in early breast cancer.
Authors: Jung S , Han W , Lee J , Yu J , Ko E , Kim E , Moon H , Park I , Hwang K , Oh D , Kim T , Noh D
Published: December 2010

Abstract #1086

Purpose: We sought to determine the significance of Ki-67, one of the tumor cell proliferation indexes, as a useful prognostic factor in early breast cancer.  Patients and Methods: 1,080 consecutive patients with stage I or II breast cancer operated between 1998 and 2003 were enrolled. Patients were categorized based on the 2007 St. Gallen consensus and Adjuvant! Online. The expression of Ki-67 in the tumor was assayed using immunohistochemistry (cut-off value: 10%).  Results: Univariate analysis revealed that tumor size, lymph node involvement, histologic grade, estrogen receptor, progesterone receptor, bcl-2, and Ki-67 (≥10%) were significant for both overall survival (OS) and distant metastasis-free survival (DFS). Of them, lymph node involvement and high Ki-67 expression were identified as independent prognostic factors for OS at multivariate analysis. The survivals of intermediate- and high-risk groups according to 2007 St Gallen consensus were further separated by Ki-67 expression level (5-yr DFS rate=93.3% vs 86.6% for Ki-67<10% and ≥10%, respectively in intermediate-risk group (p=.001); 5-yr DFS rate=83.1% vs 61.5% for Ki-67<10% and ≥10%, respectively in high-risk group (p=.006)). The survivals of low- and high-risk groups according to Adjuvant! Online were further separated by Ki-67 expression level [5-yr DFS rate=97.8% vs 89.5% for Ki-67<10% and ≥10%, respectively in low-risk group (p=.010); 5-yr DFS rate=90.4% vs 82.6% for Ki-67<10% and ≥10% in high-risk group (p=.005)).  Conclusion: Ki-67 was an independent prognostic factor for DFS and OS in early breast cancer, and could give additional prognostic information on the risk grouping by 2007 St Gallen consensus and Adjuvant! Online.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1086.
Vascular endothelial growth factor receptor 2 is a significant negative prognostic biomarker in triple-negative breast cancer: results from a controlled randomised trial of premenopausal breast cancer.
Authors: Ryden L , Ferno M , Stal O , Linderholm B , Ostman A , Jirstrom K
Published: December 2010

Abstract #1087

Background: Triple-negative breast cancer (TNB) is today an established phenotype of primary breast cancer recognized by estrogen (ER) negativity, progesterone (PgR) negativity and (human epidermal growth factor) HER2 negativity. The prognosis is poor and moreover patients with TNB dont benefit from established targeted drugs with antiestrogen therapies or trastuzumab. The role of other putative targets in TNB, like angiogenic markers and additional tyrosine kinase receptors is therefore a topic of interest in order to find effective targeted therapies improving the prognosis.  Patients and methods: In a cohort of premenopausal breast cancer (n=564) randomized to adjuvant tamoxifen or no adjuvant treatment a tissue microarray was prepared from available primary tumors (n=500) and immunhistochemical (IHC) staining was performed including ER, PgR, HER2, vascular endothelial growth factor A (VEGF-A), vascular endothelial growth factor receptor 2 (VEGFR2), epidermal receptor growth factor (EGFR) and platelet derived growth factor ß (PDGFR ß). HER2 status was additionally defined using fluorescence in situ hybridisation (FISH). Data regarding Nottingham histological grade (NHG) and Ki67, node status, tumour size and age was also included. All included patients were used for descriptive analysis, whereas only untreated patients were included in survival analysis.  Results: In the whole cohort 96 patients had a TNB phenotype characterized by low age, high NGH (p<0.001), high Ki67-index (p<0.001), T2 tumors (p=0.002), N-negativity (p=0.004. EGFR-positivity (p<0.001) and tumor-specific VEGFR2 expression (p=0.03). TNB phenotype was not associated with VEGF-A or stromal expression of PDGFR ß, and was a significant prognostic factor for 10-year BCSS (HR 2.1 CI: 1.4-3.3) by univariate analysis in the untreated arm. The prognosis was further explored among TNB patients and VEGFR2 was a significant negative prognostic factor (HR 2.5 CI; 1.1-5.5) for 10-year BCSS, whereas VEGF-A , EGFR and PDGFR ß were not.  Discussion: In a controlled randomized trial of premenopausal breastcancer, TNB was associated with worse prognosis. VEGFR2 was a significant prognostic biomarker for TNB patients which makes VEGFR2 a putative therapeutic target for this specific phenotype. Additional analysis of EGFR, VEGF-A and PDGFR ß did not add any prognostic information for TNB patients in this study.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1087.
HER2 status determined on pre operative biopsies and in operative specimens: concordance among immunohistochemistry (IHC) fluorescence in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) techniques in the CRITHER study.
Authors: Arnould L , Roger P , MacGrogan G , Lannes B , Hayem C , Campana F , Balaton A , Penault-Llorca F
Published: December 2010

Abstract #1089

Background: In breast cancer, HER2 status is generally assessed by IHC or FISH on operative tissue sample and in situ hybridization techniques are standard tools for determining equivocal IHC 2+ overexpression and deciding the correct treatment for HER2-positive breast cancer patients. For the management of their patients, oncologists or surgeons sometimes ask for assessment of HER2 status on per cutaneous core biopsy (CB).The primary objective of the CRITHER study was to investigate concordance between the results of CISH, FISH, and IHC techniques on pre-operative CB samples for HER2 status compared with HER2 status determined by FISH on surgical specimen from a population of previously untreated non-metastatic breast cancer patients in France. Methods: Analysis was performed on samples from 260 patients recruited at 24 pathology centers for the period 2003 to 2006 inclusive. Recruitment was made on the basis of IHC obtained from operative tissue samples in order to obtain around 50% HER2-positive (3+), 30% HER2-negative (0/1+), and 20% HER2-equivocal (2+) cases. CISH and IHC were performed in parallel on the pre-operative biopsy and matched surgical specimen in the participating laboratories, and FISH on both specimens in each of four reference pathology centers. IHC staining was analyzed according to different scoring systems. Results: The rate of discordance between pre-operative CISH and operative FISH was 1.8% with 1.5% false-positives and 2.2% false-negatives (k=0.963; p<0.001); the corresponding rate for pre-operative IHC (excluding equivocal IHC 2+ cases) and operative FISH was 1.5% with 1.6% false-positives and 1.4% false-negatives (k=0.953; p<0.001). Comparison of pre-operative and operative samples showed expected low rates of discordance for CISH (0.85%; k=0.982; p<0.001) and FISH (0.45%; k=0.991; p<0.001), with no false positives in either case. Results for the IHC scoring system analyses on concordance will be presented at the meeting. Discussion: As expected, concordance for both CISH and FISH between pre-operative and operative results was excellent. There was a high rate of concordance for pre-operative CISH and IHC HER2 status determination on CB (excluding equivocal IHC 2+ cases) compared with operative FISH results in patients with previously untreated non-metastatic breast cancer.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1089.
Meningeal carcinomatosis in patients with breast cancer: clinical features and prognostic factors in 91 patients from a single institution.
Authors: Gautier H , Guilhaume M , Bidard FC , Dieras VC , Pierga J , Livartowski A , Mignot L
Published: December 2010

Abstract #1090

Background  Breast cancer is the leading solid tumor responsible for meningeal carcinomatosis (MC) and the incidence is rising. To date, there is no standard of care and few prospective trials. Intrathecal treatment is provided but the modalities remain controversial.  Methods  From our data base we recorded 91 patients with breast MC admitted to Institut Curie between 2000-2007. Lumbar punction was performed in all patients for cerebrospinal fluid (CSF) cytology, protein level and dosage of CYFRA 21. Intra-thecal treatment: methotrexate 15 mg day1 to 5 and hydrocortisone acetate day 1 with oral folinic acid rescue day 1 to 5 every two weeks.  Results  Median age was 53 years (range 30-78). Tumor type was ductal carcinoma (63%), lobular (28%), ER positive 74%, HER2 overexpressed 10% and triple negative 31%. All patients presented other metastatic site: bone (67%), liver (42%) lung (29%), brain (38%). The median interval between primary tumor and diagnosis of MC was 60 months.  Amongst the 80 patients treated (87%) with intra-thecal MTX, clinical response was observed in 88% of cases. There was a strong correlation between clinical response and survival (p<0.001). Normalization of cerebrospinal fluid cytology (CSFC) was observed in 13 % of patients and was associated with better survival: 13 months versus 2 months (p=0.003). Improvement of other biologic parameters as CSF protein and CYFRA 21 was observed in 17% and 80% of patients respectively. The median survival was 4.5 months (0-53). In multivariate analysis, prognostic factors were: performance status 3-4 (p<0.0001), more than 3 chemotherapy regimens before MC diagnosis (p=0.04), lack of ER (p=0.04) and high CYFRA 21 at diagnosis (p=0.01). With these factors a prognostic score may select patients with a true benefit to IT MTX (20% survival > 1 year) or patients for whom treatment will have no impact on survival (< 3 months).  Conclusion: This score has to be confirmed in prospective study and new modalities of treatments need to be evaluated according to the biology of disease.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1090.
Men attending a breast cancer risk evaluation clinic.
Authors: Vaz FH , Opinião AS , Rodrigues PC , Machado PM , Fragoso S , Santos S , Silva J , Moura C , Oliveira J
Published: December 2010

Abstract #1103

Background- Counselling for hereditary breast cancer risk has been mostly directed to women and their female counterparts. However, BRCA1/2 male carriers are at risk for several cancers and their motivation for counselling and follow up is mostly unknown.  Material and methods- Review of individual and family records from all male pts counseled in our clinic from Jan2001-Dec2007.  Results: eighty one male pts were counseled, 32 previously affected with cancer (mean age 61yrs) and 49 healthy men (mean 41 yrs). Mostly, all affected pts were refered to or invited to as probands for BRCA2 screenning. All but 2 of these accepted screening after counseling, being the concern with their offspring the main motivation for testing. With 3 exceptions all unaffected men were unvited for counselling due to the identification of BRCA2 mutations in their families: 6/49 refused or postponed genetic testing after counselling (mean age 39 yrs). All male BRCA carriers (total of 32) are BRCA2 positive: 12 index probands affected with Breast (10 cases, 3 bilateral) or prostate cancer ( 2 cases) and 20 healthy carriers under increased surveillance (yearly observations at the Breast Clinic + Dermatology and Urology). Psychosocial support is also available but only one male BRCA2 carrier, a prostate cancer survivor with previous depressive symptons, has this support regularly.  Male carriers did adhere to increased surveillance and 5 neoplasias were diagnosed in 4 BRCA2 male breast cancer survivors: 3 cases of asymptomatic prostate cancer (Gleasons 9, 7 and 6 respectivly) 1 case of contralateral breast cancer and 1 case of gastric cancer. All 4 pts are alive without evidence of disease, after appropriate treatment. Conclusions: men belonging to BRCA families are at risk for several cancers and seem to adhere to genetic testing and surveillance. Guidelines for hereditary breast cancer management should be comphreensive and include orientations to counsell male individuals at risk.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1103.
Identification of urinary biomarkers to distinguish tumor bearing and control rats in the methylnitrosourea (MNU) – induced model of mammary carcinogenesis: use of label-free, comparative, ultra-high resolution nano-LC mass spectrometry.
Authors: Townsend RR , Rohrs H , Leduc R , Yi Y , Yao R , Malone JP , Edrman-Gilmore P , Grubbs CJ , You M , Lubet RA
Published: December 2010

Abstract #1109

In the MNU induced model of mammary cancer, rats given a single dose of MNU (75 mg/kg BW), via the jugular vein, at 50 days of age develop multiple ER+ mammary cancers. These cancers appear similar to highly differentiated ER+ human breast cancers, and have been shown to be sensitive to many of the same chemotherapeutic agents that are effective in women. When a female rat developed its first palpable cancer, it was placed in a metabolic cage and urine collected overnight on dry ice. The urine from these rats were compared to those obtained from control rats. Using a new sample preparation protocol, peptides were prepared from the urines of control (n = 4 individual animals) and rats bearing induced mammary tumors (n = 4). The peptide pools were analyzed using nano-LC-LTQ-FT-MS. The 8 LC-MS analyses were aligned and the integrated accurate mass signals were quantified using Rosetta ElucidatorTM software. The ion currents from 4430 peptides (charge state > +2) were analyzed between the two groups using ANOVA. Two hundred and twenty nine peptides were found to be significantly different (P< 0.01) between the two groups, with clear distinction between the urines from control and experimental animals being observed using hierarchical clustering and principal component analysis. The protein identifications that were significantly increased in the tumor-bearing urines were confirmed by targeted tandem mass spectrometry. Further studies relating the altered expression of these peptides to differences in tumor size, the effects of therapeutic agents, etc will also be presented.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1109.
Preventive effects of the EGFr inhibitor erlotinib in ER+ methylnitrosourea (MNU) induced and in ER- MMTV/Neu/P53KO mammary cancer models.
Authors: Grubbs CJ , Iwata KK , Buck E , Juliana M , Lubet RA
Published: December 2010

Abstract #1112

The preventive/therapeutic efficacies of Erlotinib in both ER+ and ER- models of breast cancer in rats and mice, respectively, were evaluated. For the ER+ model, female Sprague-Dawley rats were administered MNU at 50 days of age. Beginning five days later, Erlotinib was administered 7x/week for the remainder of the study. Daily administration of Erlotinib in a prevention setting at 4 and 1.33 mg/kg BW/day reduced tumor multiplicity by 72 and 35%, respectively; doses >6 mg/kg BW/day decreased cancers by >90%. When rats with palpable mammary cancers were treated for six weeks with 4 mg/kg BW/day of Erlotinib, most of the tumors showed highly significantly regressions. Similarly, when rats bearing palpable mammary cancers were treated with Erlotinib for 5 days (3 or 1 mg/kg BW/day), large decreases in tumor cell proliferation and alterations in expression of phosphorylated EGFr, AKT, ERK and SHC were observed. In additional studies, the preventive effect of Erlotinib in a MMTV/Neu/P53KO bitransgenic model was examined. These mice develop ER- cancers which overexpress Neu and have an alteration in P53. These changes are characteristic of ER- human breast cancers. An initial experiment showed that Erlotinib (50 mg/Kg BW/day) was also effective as a preventive agent in this model. Effects of this agent on proliferation and altered expression of phosphorylated EGFr, AKT, ERK and SHC are in progress. Our results showing efficacy of the EGFr inhibitor Erlotinib in ER+ mammary cancers are in agreement with a clinical study of early stage human ER+ breast cancer showing that the EGFr inhibitor Gefitinib is highly effective in ER+/EGFr+ tumors (Polychronis, et al., Lancet Oncolo.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1112.
Expression profile of CYP19 in human breast cancer cell lines treated with indole-3-carbinol, 3,3'-diindolylmethane, and sulforaphane.
Authors: Licznerska BE , Matuszak I , Baer-Dubowska W
Published: December 2010

Abstract #1113

Breast cancer is the most frequently diagnosed cancer in Western women. Most cases (up to 75%) are estrogen-dependent. The estrogen-independent cases are more invasive, more difficult to treat, and they are connected with worse prognosis. The biologically active estrogens important in the proliferation of breast cancer are produced from inactive precursors by the action of aromatase encoded by CYP19 gene. Conventional treatment (e.i. hormonal therapy) is expensive and for advanced cancer ultimately unsuccessful. That is why early diagnosis and prevention of this disease is urgently needed. A new hope is brought by chemoprevention – the concept of early intervention by usage of different agents inhibiting the development of invasive cancer. Recent epidemiological migrant studies showed that consumption of raw or short cooked cabbage and sauerkraut is connected with significant reduction of breast cancer incidences. Indole-3-carbinol (I3C), 3,3'-diindolylmethane (DIM) – a major in vivo acid-catalyzed condensation product of I3C, and sulforaphane (SUL) are examples of numerous active chemicals in cabbage.  The aim of the present study was to investigate the effect of I3C, DIM and SUL on the profile of CYP19 expression in human breast cancer estrogen-dependent MCF-7 cell line. Cells were treated with these compounds at the concentrations relevant to those observed in human plasma. The total RNA extraction from cell cultures after 144 hours of incubation with different substances concentrations and reverse transcriptase reaction preceded the cDNA synthesis. The screening of cDNA from aromatase mRNA was performed using real-time PCR assay. The results showed that I3C in the dose of 30 μM reduced the expression of CYP19 in MCF-7 cell line. In contrast, higher dose of I3C (50 μM) induced its expression. The latter results confirmed the observations of the other authors indicating reciprocal I3C effect on carcinogenesis. SUL in the doses of 5μM and 20μM diminished the level of CYP19 mRNA. Similar trend was observed after treatment with 5 μM and 10 μM of DIM. Since the down-regulation of CYP19 gene causes the reduction in estrogens synthesis the usage of I3C, DIM, and/or SUL could be well-founded as preventive agents against breast carcinoma development. Further investigation on estrogen-independent MDA-MB-231 cell line which is under way are necessary to compare the possible application of I3C, DIM, and SUL as a chemopreventive agents depending on the hormonal character of the tumour.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1113.
Treatment of early breast cancer in women over 65: what is important? Retrospective trial.
Authors: Kim K , Jung Y , Hur L , Kim S
Published: December 2010

Abstract #1125

Introduction-Background-Objective  There are some demonstrations for the under-treatment of elderly (age 65 or older) women with early breast cancer. However, a few reports are available for overall survival as well as treatment patterns. To characterize treatment patterns and to determine the impact of under-treatment of early breast cancer in the elderly population compared to younger population.  Methods  We analyzed data of 4,990 early (stage I, II) breast cancer patients aged ≥ 50 years old postmenopausal women, registered in the Korean Breast Cancer Society Registration Program (KBCSRP) between 1992 and 2001 retrospectively. We compared elderly group (age 65 or older) with younger group (age 50 or older, and under age 65) in cancer stage, hormonal receptor status, p53 status, HER-2 status, breast conservation rate, radiation therapy omission rate after breast conservation surgery, chemotherapy rate, hormonal therapy rate and overall survival.  Results  942(19%) patients were in the elderly group and 4,048(81%) patients were in the younger group. We followed up to 2004 and median follow up period were 5.7 years. There were no differences in cancer stage, HER-2 status, and hormonal therapy rate.  In elderly group, tumor had two better prognostic factors: more hormonal receptor positivity (63.3% vs. 59.3%; p=0.001) and less p53 positivity (37.3% vs. 49.5%: p<0.001). Overall survival, however, was lower than younger group: 83% vs. 91% in 5 year survival rate, 68% vs. 81% in 10 year survival rate, respectively. Elderly group had less breast conservation rate (16.4% vs. 20.4%; p<0.001), more radiation omission rate after breast conservation surgery (59.8% vs. 15.3%; p<0.001) and less chemotherapy rate (43% vs. 70.6%; p<0.001). Using log-rank analysis, overall survival was affected by under-treatment of disease: women who underwent breast conservation surgery without radiation therapy experienced a significant decreased survival compared to women with radiation therapy (68% vs. 84% in 10 year survival rate; p<0.001). Total mastectomy was not superior to breast conservation surgery plus radiation therapy (p=0.2856). Women who underwent chemotherapy, however, revealed no survival gains, but women with hormonal therapy showed superior survival rate (p=0.0019).  Conclusions  Our data revealed that elderly patients had better prognostic tumor characteristics, but lower survival rate. Deviated under-treatment patterns in elderly early breast cancer patient could be dangerous choice to patients, therefore, radiation therapy after breast conservation surgery should not be omitted and hormonal therapy should be recommended especially.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1125.
Phase III randomized adjuvant study of tamoxifen alone versus sequential tamoxifen and anastrozole in postmenopausal women with hormone-responsive breast cancer: N-SAS BC03 study.
Authors: Aihara T , Takatsuka Y , Osumi S , Aogi K , Hozumi Y , Imoto S , Mukai H , Iwata H , Watanabe T , Shimizu C , Nakagami K , Tamura M , Ito T , Masuda N , Ogino N , Hisamatsu K , Mitsuyama S , Abe H , Yamaguchi T , Ohashi Y
Published: December 2010

Abstract #1138

Background. Clinical trials conducted in Western countries have shown that improved disease-free survival (DFS) is associated with the use of aromatase inhibitors instead of tamoxifen in postmenopausal women with early breast cancer. Pharmacogenetic differences in drug metabolizing genes may cause ethnic differences in the response or tolerability to aromatase inhibitors, as well as tamoxifen. In fact, improved DFS by extending the use of letrozole after tamoxifen for 5 years has not been confirmed in women from minority groups (Ann Oncol. 2006 17(11):1637-43). Therefore, assessment of the efficacy of aromatase inhibitors in non-white women is warranted. We report the results of switching adjuvant therapy from tamoxifen to anastrozole in postmenopausal Japanese women with breast cancer enrolled in an open-label, randomized clinical trial (N-SAS BC03 study). Patients and Methods. From November 2002 to December 2005, 706 postmenopausal women with hormone-receptor-positive breast cancer who were receiving adjuvant therapy with tamoxifen for 1 to 4 years were randomly assigned to either continue tamoxifen or to switch to anastrozole; the total duration of treatment was 5 years. Primary endpoints were DFS and adverse events. Secondary endpoints included relapse-free survival (RFS), overall survival (OS) and QOL. Results. After a median follow-up of 42 months (range, 3.2-60), there were 26 events in the anastrozole group as compared with 37 in the tamoxifen group related to DFS, and 15 events in the anastrozole group as compared with 28 in the tamoxifen group related to RFS. The unadjusted hazard ratio was 0.69 (95 percent confidence interval, 0.42 to 1.14; P=0.06 by the log-rank test) for DFS and 0.52 (95 percent confidence interval, 0.28 to 0.98; P=0.02 by the log-rank test) for RFS, both in favor of anastrozole. There was no difference in OS (p=0.49). Conclusion. Switching from tamoxifen to anastrozole was confirmed to be associated with a lower disease recurrence rate than continuing tamoxifen in postmenopausal Japanese women with breast cancer.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1138.