bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic Significance of Placenta Growth Factor (PlGF)-1 and PGF-2 Expression in Human Breast Cancer.
Authors: Escudero-Esparza A. , Martin T. , Mansel R. , Jiang W.
Published: December 2010

Background: Placenta growth factor (PlGF) is a member of the vascular endothelial growth factor (VEGf) family, a group of angiogenic growth factors important in cancer development. Over-expression of PlGF is known to be associated with pathological angiogenesis. Recently, PlGF has been shown to have 4 isoforms, -1, -2, -3 and -4. This current study examined the expression of PlGF-1 and -2 in human breast cancer and how changes in expression of these isoforms may be linked to prognosis of the disease.Materials and Methods: Breast cancer primary tumours (n=114) and matched background tissue (n=30) were processed for RNA extraction. RNA was reversed transcribed and quantified before analysis by Q-PCR. The results were expressed as copy number of transcript/50ng RNA and standardised using β-actin. Statistical analysis was carried out using student t test (mean±SD) and Mann-Whitney U test (median/IQR) where appropriate. Immunohistochemistry was also carried out on a number of matched background and tumour sections.Results: Analysis of patients with a 10 year follow-up showed that transcript levels of PlGF-1 were increased in node positive tumours when compared to node negative (104.2±52.2 versus 52.5±13.6, respectively, but this did not reach significance(p=0.34). Prognostic indicators (NPI status, TNM status and grade of tumour) were not associated with significant changes in levels of PlGF-1. Moreover, there was no difference in expression with overall ER status of tumours; however, PlGF-1 was significantly increased in ERβ positive tumours (median values ERβ-ive and ERβ+ive (14.10 vs 68.75 respectively, p=0.014). Most interestingly, there was a significant increase in PlGF-1 expression in patients with poor outcome (median values patients remaining disease-free 16.5, patients who died from breast cancer 78.2, p=0.03). In comparison, the PGF-2 isoform showed little change in expression between tumour and background samples. However, Immunohistochemistry demonstrated a marked increased in PGF-2 expression in tumour sections (staining intensity (mean±SD): Background 102.05± 46.65 vs Tumour 172.85±40.33, n=20, p=0.0001).Conclusions: We can conclude from this data that there is a differential expression of the PlGF isoforms in human breast cancer. Inappropriate expression of PlGF isoforms may play a role in human breast cancer progression.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3041.
Association of Proliferating Macrophages with High Grade, Hormone Receptor Negative Breast Cancer.
Authors: Tonlaar N. , Campbell M. , Garwood E. , Khramtsov A. , Moore D. , Au A. , Baehner F. , Huo D. , David M. , Oluwasola O. , Odetunde A. , Tretiakova M. , Li S. , Gong C. , Tonner E. , Fridlyand J. , Falusi A. , Mcgrath M. , Gray J. , Olopade O. , Esserman L.
Published: December 2010

Macrophages, a key cell in the inflammatory cascade, have been associated with poor prognosis in cancers, including breast cancer. Tumor associated macrophages (TAM) have also been shown to play a role in invasion and metastases. In this study, we investigated the role of a subset of macrophages known as proliferating macrophages (promacs) in breast cancer. We examined the relationship between promacs and clinico-pathologic characteristics such as tumor size, grade, lymph node metastasis, hormone receptor status, molecular subtype, and survival. This study was conducted at two independent institutions (University of California, San Francisco and University of Chicago) using two independent cohorts of patients with breast cancer. Formalin-fixed, paraffin-embedded sections and/or tissue microarrays were double-stained with anti-CD68 (a macrophage marker) and anti-PCNA (a proliferation marker) antibodies. The number of total macrophages as well as promacs per high power field were counted. Molecular subtypes were determined from gene expression array analyses. Outcomes were available for a subset of the cases and Kaplan Meier survival curves were generated.Tumor tissue from two different cohorts of patients with breast cancer were stained and analyzed at two independent institutions. In both studies, promac density was significantly correlated with higher grade, hormone-receptor negative tumors, and a basal-like subtype. In contrast there was no correlation between numbers of promacs and tumor size, stage, or number of involved lymph nodes. The presence of increased promacs was a significant predictor of survival in these treated patients. Proliferating macrophages are more abundant in high grade tumors, and predict a worse outcome, independent of stage and grade. These findings, corroborated at two independent institutions, suggest that the presence of promacs is associated with tumor progression and increased promac density may serve as a prognostic indicator for poor outcomes. Novel therapies that are able to target and halt the activity of promacs may be a promising strategy for the treatment of promac rich tumors.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3042.
Adverse Clinico-Pathological Features in Breast Cancer Patients under the Age of 35.
Authors: Babar M. , Garvin J. , Quinn L. , Dunne B. , Griffin M. , Kennedy J. , Boyle T. , Connolly E.
Published: December 2010

IntroductionBreast cancer in younger patients is a unique disease entity, associated with aggressive biologic phenotype and poorer prognosis. The aim of this study was to analyse age as a prognostic factor in pre-menopausal breast cancer specifically in regards to pre-menopausal patients under the age of 35.MethodsA 7 year prospective audit of pre-menopausal patients presenting to the symptomatic breast clinic in St. James's Hospital between 2001 and 2008 was conducted. The Clinico-pathological features of pre-menopausal patients presenting under the age of 35 versus over 35 where compared. All cancer diagnoses were discussed at the breast multidisciplinary conference, and all clinico-pathological treatment details and follow-up information were entered by a full-time data manager.ResultsPre-menopausal patients under the age of 35 (n=55) were more likely to present with higher grade (p<0.05), oestrogen receptor negative (p<0.05), HER-2 receptor negative (p<0.05) and stage IV disease (p<0.05) relative to the older premenopausal group (n=250). This group also had a non-significant increased rate of lymphovascular invasion and negative progesterone receptor status compared to the older premenopausal group (p> 0.05). Pre-menopausal patients over the age of 35 were twice as likely to have lobular carcinoma as opposed to their younger counterparts. Tumour size, nodal status and axillary node burden was not significantly different between groups. Patients under-35 with breast cancer were more likely to be nulliparous.ConclusionThis data demonstrates that breast cancer in pre-menopausal patients under the age of 35 is associated with adverse clinicopathological features relative to the older pre-menopausal patients. Further studies should be aimed at assessing the prognosis, disease recurrence, disease free survival and distinct genetic profile in age related breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3043.
Metabolic Profile (Blood Pressure, Lipids and Body Mass Index) in Relation to Breast Cancer Mortality.
Authors: Emaus A. , Veierød M. , Tretli S. , Finstad S. , Selmer R. , Furberg A. , Bernstein L. , Schlichting E. , Thune I.
Published: December 2010

Background: Metabolic profile (body mass, blood pressure, lipids) may affect biological mechanisms of importance for breast cancer. In the present study we have studied whether these factors are related to breast cancer mortality.Material and Methods: Within the Norwegian Counties Study, a population-based study (1974-2005), we identified 1364 female invasive breast cancer cases.Results: The 1364 cases had a mean age at diagnosis 57.5 years (range 27.1-79.4 years), mean body mass index (BMI) was 24.0 kg/m2, mean systolic blood pressure was 128.1 mmHg, mean diastolic blood pressure 78.6 mmHg, mean serum cholesterol was 6.17 mmol/L and mean HDL-cholesterol was 1.45 mmol/L. A total of 429 cases died during a mean follow-up of 8.2 years. Those with a BMI ≥30 kg/m2 had a 1.45 (CI, 1.07 to 1.95) higher risk of dying during the follow-up than women with a BMI of 18.5–25 kg/m2. Women in the highest tertile of blood pressure had a 43% increase in mortality compared to women in the lowest tertile of blood pressure (HR=1.43, 95% CI, 1.10 to 1.85). Additionally, women in the highest tertile of total cholesterol had a 29% increase in mortality compared to women in the lowest tertile (HR=1.29, 95% CI, 1.01 to 1.65).Conclusion: Our study supports a relationship between total mortality among breast cancer patients and metabolic profile.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3047.
Comparative Validation of the SP6 and MIB1 Antibodies to Ki67 and Their Use in Tissue Microarray (TMA) and Image Analysis for Breast Cancer.
Authors: Zabaglo L. , Zabaglo L. , Salter J. , Salter J. , Anderson H. , Anderson H. , Hills M. , A'Hern R. , Dowsett M. , Dowsett M.
Published: December 2010

Aim: To compare SP6 and MIB1 antibodies against Ki67 for use with visual scoring, image analysis and TMA (tissue microarray) analysis.Background: Immunohistochemical detection of Ki67 has been widely used in assessing the proliferation fraction in breast cancer. Neoadjuvant studies revealed that changes in Ki67 predict benefit from endocrine treatment, and that on-treatment Ki67 measurements are better predictors of long-term outcome than pretreatment levels (Dowsett at al, JNCI 2007; 99:167-170). As a result Ki67 is the primary end-point for some pre-surgical trials. MIB-1 has been the preferred Ki67 antibody for over 10 years. A rabbit monoclonal antibody, SP6, has become available with apparently improved performance, e.g. reduced background non-nuclear staining. The importance of Ki67 led us to conduct a systematic series of studies to validate SP6 in comparison to MIB1.Methods: We used TMAs constructed from (i) 177 archival FFPE blocks of primary or locally recurrent breast tumours from women treated with an aromatase inhibitor (AI) for advanced disease (AI TMAs) and (ii) one set of replicate blocks (200 tumours) from the TransATAC program. The TMAs were of 600µm cores. Three replicate TMAs (A, B, C) were used. Staining was performed using DAKO Autostainer and DAKO REAL Kit Detection System. MIB-1 antibody (DAKO) was used at a dilution of 1:40, whereas 1:100 dilution was used for SP6 antibody (Abcam). Stained sections were examined either visually or using the Ariol image analysis system (Genetix Ltd.).Results: There was a strong correlation between the antibodies in AI TMAs scored manually (n=161, r=0.93, p<0.0001). Coefficient of variation was calculated in all cases with ≥2 available cores. MIB1 showed higher median CV [%] compared with SP6 (43.7 vs. 36.7, respectively) in AI TMAs, but a lower CV in TransATAC TMAs (42.0 vs. 51.4, respectively). In a univariate analysis of manual scores from AI TMAs there was a similar association of Ki67 with time to treatment failure (TTF) with the two antibodies (HR: 1.15; 95% CI: 1.00-1.33; p=0.052 and HR: 1.19; 95% CI: 1.00-1.43; p=0.045 for MIB1 and SP6 respectively). Correlation between manual and image analysis scores was markedly better with the SP6 antibody (r=0.71 and r=0.88 for MIB1 and SP6, respectively).Conclusions: SP6 and MIB1 provide highly comparable measures of Ki67 that predict progression of advanced disease similarly. SP6 is substantially better suited than MIB1 to image analysis, and is now our preferred antibody for future studies.Funded by Breakthrough Breast Cancer and The Mary-Jean Mitchell Green Foundation.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3048.
Outcome of Isolated Sternal Recurrence in Breast Cancer.
Authors: Astrow A. , Desani J. , Sonnenschine M. , Patsiornik Y. , Huang Y. , Klein P. , Citron M. , Reichman B.
Published: December 2010

Introduction: Metastatic breast cancer to sternum is rare and its natural history and prognosis are not fully understood. Metastasis to sternum is considered as stage IV, but may represent local extension from internal mammary nodes. Here we present a case series of 7 patients with metastatic breast cancer to sternum with and without internal mammary node involvement.Method: Seven patients were identified by 5 medical oncologists from New York area with busy breast cancer practices, from 1991 to 2008. Patients chart were reviewed and information regarding age, menopausal status, hormone receptor status, Her2/neu status, initial treatment, time to sternal recurrence and treatment of metastatic disease were obtained.Results: Out of 7 patients with stages I to III at diagnosis, 4 were premenopausal. Two patients had triple negative tumors, 3 were ER/PR positive and 2 were Her2 positive. All patients received loco-regional treatment of the primary site with surgery and/or radiation. Five patients received adjuvant or neoadjuvant chemotherapy. Median time to sternal recurrence was 3 years. Two patients had biopsy confirmed metastasis to internal mammary nodes at relapse. All patients with recurrence received either chemotherapy, radiation or hormonal therapy. One patient received autologous bone marrow transplant after sternal recurrence. All 7 patients are alive, 5 of them in remission and 2 patients with stable disease. Median DFS in 5 patients with CR was 7.8 years.Conclusion: Isolated sternal recurrence with/without internal mammary node involvement is relatively uncommon and is classified as stage IV. On our review of 7 cases, 5 remain in continuous remission at a median of 8 years from recurrence. The other two patients have stable disease. These sternal lesions could possibly represent loco-regional recurrence rather than systemic relapse. Hence, isolated sternal recurrence from breast cancer may need to be stratified separately in studies of stage IV disease.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3054.
Genomic Profiling of Breast Cancer (BC) in Mali by Oligonucleotide Comparative Genomic Hybridization (CGH) Array.
Authors: Ly M. , Diallo G. , Valente A. , Diallo D. , Dumke K. , Marty V. , Scott V. , Pusztai L. , Berrada N. , Lazar V. , Job B. , Viehl P. , Bernaudin J. , André F.
Published: December 2010

Background: Epidemiological studies suggest that BC in Africa exhibit specific biological characteristics as compared to BC from US/Europe. In order to identify therapeutic targets that would be specific from Africa, we analyzed 28 samples from Mali by high resolution CGH array, and compared gene gains/losses to the ones observed in USA [André F et al, Clin Cancer Res, 2009]Patients and methods: 71 paraffin-embedded samples of invasive BC from patients treated by surgery at Bamako were retrieved, 28 of them (from 2 pts stage 1; 13 pts stage 2; 11 pts stage 3 and 2 pts stage 4) were eligible for CGH array (1) (Agilent, 4*44K) (cancer cells>50%; undamaged DNA >1µg).Results were compared to 106 samples from MD Anderson, profiled in the sample genomic platform using the same array (1)Results: The median age was 49 year old. Twenty-one samples (75%) were ER-negative. No difference was observed regarding the frequency of gene gains in 17q12-21 (ERBB2) nor 8p11-12 (FGFR1). Six regions were more frequently gained in BC from Mali, as compared to those from US (FDR<5%). Two regions included targetable molecular abnormalities. 9p34 region was gained in 43% of samples and included Abl1, a tyrosine kinase involved in CML, and VAV2, a guanine exchange factor. 22q11-13.1 region was gained in 34% of samples and included bcr, ewsr1, pdgfb and MAF-E. These two regions were gained in less than 5% of BC from US. Finally, a region located in 6p21 was gained in 56% of BC from Mali. This region included a cluster of genes involved in immune response (lymphotoxin A, MICA/MICB).Conclusion: BC from Mali present some candidate specific targetable alterations as gains of Abl1, bcr and pdgf-B. FISH and IHC are being done in order to validate targets.Additional high throughput analyses will be performed to characterize such tumors, including virus arrays, exon arrays and hot spot mutations.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3059.
Screening of the Portuguese BRCA2 Founder Mutation in Breast Cancer Patients Unselected for Family History.
Authors: Opinião A. , Opinião A. , Fragoso S. , Lourenco P. , Sousa P. , Braga S. , Braga S. , Luís A. , Luís A. , Machado P. , Rodrigues P. , Vicente H. , Oliveira J. , Bruges-Armas J. , Vaz F. , Vaz F. , Vaz F.
Published: December 2010

Background and Objective: The identification of BRCA2 c.156_157insAlu as a founder mutation of Portuguese origin and the optimization of its screening method, allows for fast genetic diagnosis in positive families. Although prevalent in Portuguese families with hereditary breast cancer risk (45% of the families with identified mutations in our Institute are positive for BRCA2 c.156_157insAlu), its prevalence in the general population of Portuguese breast cancer patients unselected for family history is unknown. Our objective was to determine the frequency of c.156_157insAlu in general Portuguese breast cancer (BC) population.Material and methods: After Ethics approval, recruitment started in April 2008.Two centres started the study: the Cancer Institute in Lisbon that observes patients from the Centre and South of the country, and the Hospital of Terceira Island in Azores (patients recruited in Terceira Island were originated from all Azorean Islands). All these patients had the diagnosis of invasive breast cancer, and after counselling a peripheral blood sample was collected and BRCA2 c.156_157insAlu screened by a three-step PCR method (Machado PM et al; J Clin Oncol 25:2027-2034, 2007). Post-test counselling was given at the Breast Cancer Risk Evaluation Clinic, where all patients had their family history analyzed. If at high risk for hereditary cancer, women were advised to enter general screening of BRCA1/2 mutations, when negative for BRCA2 c.156_157insAlu.Results: Three hundred twenty three BC patients (226 from Azores; 97 from the Continent) were recruited for this study and have already been screened. Eighty-three patients from Azores met criteria for familial breast cancer (37%). In the Continent only 10 high-risk patients have been identified so far. Average age at diagnosis was 55.8 yrs; 13 patients had bilateral BC (4%); 18 (6%) patients (all from Azores) had a second cancer diagnosis. Two women (1% of all, and 2% of the patients from the Continent) were positive for the BRCA2 c.156_157insAlu mutation: 1- breast cancer diagnosis at age 45 and a maternal aunt with breast cancer at age 60; 2- bilateral BC (ages 35 and 37), no family history for breast and/or ovarian cancer. The 10 high-risk patients BRCA2 c.156_157insAlu negative accepted to proceed with general BRCA1/2 screening.Patient recruitment is still open.Conclusions: Although we cannot conclude that c.156_157insAlu screening is relevant in the unselected population, one of the positive patients did not meet criteria for counselling outside this study. Our data also suggest that, even for familial BC, c.156_157insAlu may not be as prevalent in the Azorean population.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3061.
Waiting Time for Breast Conserving Surgery Patients in a Public and a Private University Affiliated Hospital in Atlanta.
Authors: Mosunjac M. , Park J. , Strauss A. , Birdsong G. , Du V. , Rizzo M. , Okoli J. , Bumpers H. , Gabram-Mendola S.
Published: December 2010

Background: The time it takes for a patient to undergo the entire breast cancer pathway treatment varies for diverse patient populations in different health care settings. In this study we analyzed delay in breast cancer treatment (DBCT) for a defined population of breast cancer patients. All of the patients had identical cancer care trajectories of breast-conserving therapy (BCT) for infiltrating ductal carcinoma (IDC) or ductal carcinoma in situ (DCIS), followed by neo-adjuvant therapy in two different hospital settings; including a University-based inner-city hospital and a University-affiliated private practice hospital.Materials and Methods: A retrospective chart review of 214 patients treated from 2004 to 2008 was conducted. Five consecutive diagnostic and treatment events in a total of four time intervals (see Figure 1 below) were defined and correlated with demographic factors such as age, race, marital status, distance traveled to visit, insurance status type, and hospital type. Non-parametric Wilcoxon Rank-Sum test was used for statistical analysis.Results: The mean ages of the patients in both hospitals were similar (59.2 yrs public hospital vs. 61.9 yrs private hospital). Patients treated in the public hospital experienced greater DBCT compared to patients who were treated in the private institution (125 vs. 88 days, p < 0.001). Overall, the largest delay was the time period from diagnostic core biopsy to surgery (50 days). The only time interval that showed greater delay for African-American (AA) women compared to Caucasian women was time from final pathology diagnosis to medical oncology evaluations (26 vs. 33 days, p=0.036). Patients who were married or insured experienced less DBCT compared to patients who were not (80 vs. 117 days and 83.3 vs.167 days respectively, p < 0.001). Medicaid patients had shorter wait times in the public compared to in the private hospital (136 vs. 153 days, ns). Patients who experienced less than 90 days of delay to undergo the entire treatment traveled an average 27.3 miles (sd=76.6) for their hospital visit, while patients who experienced greater than 90 days of delay to undergo the same treatment traveled an average 14.2 miles (sd=19.9) for their hospital visit.Discussion: Differences in DBCT are multi-factorial and arise from system-based issues that vary among hospital settings along with demographic factors such as marital status and race that are independent of the hospital setting. More effective scheduling for surgical treatment and follow up appointments may reduce the wait time. In order to significantly shorten DBCT further, prospective studies are needed to evaluate the intricate connection of psychosocial and system barriers to breast cancer treatment.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3070.
A Multi Centre Prospective Longitudinal Study Establishing Level II Evidence of Health Related Quality of Life after Types of Immediate Latissimus Dorsi (LD) Breast Reconstruction.
Authors: Winters Z. , Mills J. , Haviland J. , Reece-Smith A. , Greenslade M. , Benson J. , Galea M. , MacMannus P. , Nicholson S. , Weiler-Mithoff E. , Rayter Z. , Thomson H.
Published: December 2010

Introduction: NICE recommends that the majority of women undergoing mastectomy should be offered immediate breast reconstruction with its potential benefits to improve health related quality of life (HRQL). A systematic review shows poor and conflicting evidence with a lack of 'hard' data to best inform both clinicians and their patients. Our aim was to evaluate the effects of implant-assisted LD (LDI) versus autologous LD (ALD) breast reconstruction on HRQL over 36 months. Recently, with the exception of the USA Breast-Q module there is a lack of a validated Breast reconstruction questionnaire.Methods: An MREC approved prospective longitudinal cohort study involving 6 centres commenced in early 2007. Serial patient reported outcome measures using the EORTC C30 (global QoL, physical functioning, fatigue, pain), BR-23 (breast and arm symptoms), 10 item Body Image Scale (BIS) and HADS, were completed pre-operatively and at 3, 6, 12, 24 and 36 months after surgery. Overall cosmetic satisfaction of the breast reconstruction was measured by a study-specific question on a 5 point Likert scale. Generalised estimating equations were used to assess change over time and differences between treatment groups.The Spearman's rank correlation coefficient was used to assess associations between patient reported cosmetic satisfaction with BIS and breast symptoms.Results: 118 patients (65 – ALD, 53 – LDI) were recruited to the study with a mean age of 50 years (range 22-70). Compliance with questionnaires at all time points was between 85-90%. There were no significant differences in HRQL domains between LDI and ALD, except a tendency for worse pain with ALD (p=0.06). Significant improvements over time were seen for overall HRQL (p<0.001), fatigue (p<0.001), breast symptoms (p=0.04), arm symptoms (p=0.001), as well as anxiety (p=0.017) and depression (p<0.001). Preliminary data shows that overall cosmetic satisfaction was significantly correlated with better body image and fewer breast symptoms. The study is ongoing in terms of 24 and 36 months follow-up and the evaluation of RT.Conclusion: There is an important need for cumulative clinical evidence in this field on which to base patient informed consent and clinical recommendations. Further analyses will assess whether there is an independent effect of RT on HRQL. A validated EORTC breast reconstruction module is in development.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3106.
Extended Indications for Skin Conserving of the Nipple Areola Complex during Subcutaneous Mastektomie.
Authors: Niemeyer M. , Rainer B. , Kiechle M. , Paepke D. , Plattner B. , Paepke S.
Published: December 2010

Background: Conventional breast cancer surgery in case of a non breast conserving procedure still includes the routine excision of the nipple areolar complex (NAC) optional followed by nipple reconstruction. Numerous authors consider multiple predictive factors to decide whether the Nipple Areola Complex (NAC) can be conserved or not during mastectomy. These are: tumour-nipple distance, tumour size, axillary lymph-node status and lymph vascular invasion. Thus only a very limited percentage of patients can keep their NAC.If the breast gland tissue and all galactophoric ducts can be separated completely from the NAC during subcutaneous mastectomy (SCM), conservation of the NAC is feasible even in large, central and retroareola tumours.Patients and methods: (www.clinicaltrials.com ID: NCT00641628) From July 2003 to Mai 2006 we performed 109 SCMs on 96 patients. 94 of these breasts showed indications for modified radical mastectomy (MRM) or ablatio simplex. 78 due to invasive carcinoma (MRM indication) and 16 of them had extensive ductal carcinoma in situ (DCIS), (indication for simple ablation). At least 33 of the breasts had malignancy underneath the skin within the areola margin (central located tumours). After dissection of the complete mammary gland tissue, the skin envelope with the areola is folded inside out an all galactophoric ducts and any subjacent tissue on the areola base are precisely dissected under the surgeons' visual control. Of this skin-closest retroareola tissue, frozen sections and HE-histopathologic examinations are requested to decide whether the NAC can be preserved or not.Results: We found the Need to dissect the NAC in 13 of 109 breasts (12%), altering the procedure to a skin sparing mastectomy (SSM). Necrosis of the NAC which had to be subjected to surgical intervention occurred in only one of the conserved 96. After a follow-up of 20-54, median 34 months no recurrence within the NAC was observed. One local recurrence on the thoracic wall and one axillary recurrence were detected. 2 of 96 patients developed distant metastases. One death was recorded. Occasionally, partial necrosis of the nipple occurred, leaving a depigmentation of the skin but maintaining a good or excellent cosmetic result in most cases. Necrosis of the NAC which had to be subjected to surgical intervention occurred in only one patient.Conclusion: SCM, including NAC conservation, may be performed according to MRM indications if an intraoperatively frozen section (and the corresponding HE-histopathology) of the NAC closest tissue is free of tumour. The remaining contraindications for SCM are: extensive tumour involvement of the skin, inflammatory breast cancer and clinical suspicious nipple.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3107.
Prolonged Injection Site Mass Can Occur with Methylene Blue but Not Lymphazurin Blue after the Sentinel Node Procedure.
Authors: Komenaka I. , Shirah G. , Bouton M.
Published: December 2010

Objectives: The purpose of this study is to evaluate the differences in outcomes and complications of the sentinel node dissection for breast cancer using Methylene blue and Lymphazurin blue.Methods: From 2004-2008, 160 patients underwent the sentinel node procedure at a county hospital, teaching institution. The sentinel node procedure was performed using the combined technique of blue dye and radioactive isotope. From 2004 to June 2006, Lymphazurin blue (LB) was used. From July 2006 to 2008, Methylene blue (MB) was used. Axillary dissection was performed if the sentinel node procedure revealed positive lymph nodes or failed.Results: Fifty patients underwent the sentinel node procedure with LB and 110 underwent the procedure with MB. The patients were of similar age and stage of presentation. More patients in the MB group were Hispanic and underwent preoperative chemotherapy. The sentinel node identification rate was 100% in the LB group and 99% in the MB group (P = 1.0). The mean numbers of sentinel lymph nodes were similar (LB 2.2 vs. MB 2.7). In the LB group 34% of patients were lymph node positive compared to 40% in the MB group.There were no allergic reactions in either group. The infection rate in the breast was similar in both groups LB (2%) and MB (1.8%). Previous studies have reported skin manifestations of necrosis, ulceration, tattooing, erythema, rash, or telangiectasia/vascular change at the site of injection with methylene blue. These findings were not seen in any of the patients in this study. In ten of the 110 patients (9%) who underwent the procedure with MB, however, a local inflammatory reaction occurred which resulted in a palpable mass at the site of injection. No injection site reactions were noted in LB group. In the MB group, the masses were clearly more prominent than the surrounding breast tissue and identified either at the one week postoperative check (6) or at the first 6-month followup (4). Ultrasound of the injection sites did reveal a hypoechoic mass in one patient. Biopsies were performed on the first four patients, but revealed only inflammation and non-atypical hyperplasia. Nearly all of the masses resolved by one year, however, one resolved after 18 months.Conclusion: Methylene blue is a reasonable alternative to lymphazurin blue in the sentinel node procedure for breast cancer. Awareness of injection site reactions to MB are important, as a new dominant mass is always reason for concern in breast cancer patients. Palpable mass at the site of MB injection can occur but typically resolve over a one year period.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3115.
Recovery of the Upper Limb Function after Sentinel Lymph Node Biopsy or Axillary Dissection.
Authors: Utsumi T. , Kobayashi N.
Published: December 2010

Introduction:Sentinel lymph node biopsy (SLNB) is widely accepted as an excellent method in the management of early breast cancer in patients with clinically negative axillary lymph nodes. The aim of this study was to evaluate the relationship between the type of breast cancer surgery and early postoperative arm mobility.Materials and methods:The study group consisted of 290 consecutive patients aged 30 to 86 years (median age 58) with breast cancer, operated upon in the Department of Breast Surgery and evaluated in Department of Rehabilitation Medicine afterward, between February 2005 and December 2008. Out of the 290 patients, 126 (43.4% - group 1) underwent breast conserving therapy with SLNB, 65 (22.4% - group 2) simple mastectomy with SLNB, 41 (14.1% - group 3) breast conserving therapy with axillary lymph node dissection (ALND) and 58 (20% - group 4) modified radical mastectomy. Shoulder mobility was evaluated after 1month and after 3 months.Results:After 1 month, patients in the ALND group (group 3 and 4) had more limitations in shoulder mobility than patients in the SLNB group (group1 and 2).After 3 months, shoulder mobility was not different between the ALND group (n=52) and the SLNB group (n=32) (shoulder flexion: 161.6±22.5 degree vs. 167.8±19.5 degree, abduction: 165.2±27.5 degree vs. 164.4±28.9 degree, respectively).Conclusion:In terms of upper limb functional status, the benefits of SLNB over ALND was observed at the early postoperative time. However, long-term effects have to be confirmed by further, larger series.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3118.
Final Results of a Randomized Phase II Study Demonstrating Efficacy and Safety of BSI-201, a Poly (ADP-Ribose) Polymerase (PARP) Inhibitor, in Combination with Gemcitabine/Carboplatin (G/C) in Metastatic Triple Negative Breast Cancer (TNBC).
Authors: O'Shaughnessy J. , O'Shaughnessy J. , O'Shaughnessy J. , Osborne C. , Osborne C. , Osborne C. , Pippen J. , Pippen J. , Pippen J. , Patt D. , Patt D. , Rocha C. , Ossovskaya V. , Sherman B. , Bradley C.
Published: December 2010

Background: TNBC is an aggressive breast cancer subtype which shares molecular and pathologic features with BRCA1-related breast cancers. BRCA-deficient cells are sensitive to inhibition of PARP1, a critical enzyme of cell proliferation and DNA repair, and thus represent a rational target of PARP inhibitor-based cancer therapy. The objectives of this study were to evaluate BSI-201, a potent PARP1 inhibitor, in combination with G/C in subjects with metastatic TNBC. Methods: Eligible subjects had measurable disease and received ≤2 prior cytotoxic regimens for ER-, PR-, and HER2-negative metastatic breast cancer. Patients were randomized (1:1) to G/C alone or G/C + BSI-201. Gemcitabine (1000 mg/m2; IV) and carboplatin (AUC = 2; IV) were given on days 1 and 8, and BSI-201 (5.6 mg/kg; IV) on days 1, 4, 8, and 11 every 21 days. Endpoints were clinical benefit rate (CBR = CR + PR + SD ≥6 months), objective response rate (ORR), progression-free survival (PFS) and overall survival (OS). Results: Preliminary analyses of all 116 dosed patients showed that, based on all patient visits through March 2009, BSI-201 + G/C improved CBR, ORR, median PFS, and median OS, compared with G/C alone. The frequency and nature of adverse events did not differ between arms. G/CG/C+BSI-201Hazard Ratio  (n=57)(n=59)(95% CI)P valueCBR, %2162 0.0002ORR, %1648 0.002Median PFS, months3.36.90.34 (0.20-0.58)<0.0001Median OS, months5.79.20.35 (0.19-0.65)0.0005 Conclusions: This analysis demonstrates that BSI-201 + G/C improved patient's outcomes, as measured by CBR, ORR, PFS and OS, compared with G/C alone. BSI-201 + gemcitabine/carboplatin was well tolerated and did not potentiate chemotherapy-related toxicities. Final data on CBR and ORR, as well as updated analyses of PFS and OS for all 116 dosed patients will be presented based on all patient visits through September 2009. Exploratory correlative analyses of PARP expression and overall clinical response will also be presented, as well as initial correlative analyses between number of metastatic therapies and clinical response.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3122.
Cisplatinum Sensitivity of BRCA1-Mutated HCC1937 Breast Cancer Cells Is Linked to Impairment of Notch Signaling and Is Increased by γ-Secretase Inhibitors.
Authors: Ventura M. , Di Martino M. , Arbitrio M. , Propato M. , Perri G. , Bulotta A. , Calimeri T. , Pacicca M. , Tassone P. , Tagliaferri P.
Published: December 2010

BRCA1 plays a critical role in DNA-damage repair mechanisms elicited by cell exposure to anti-tumor agents. HCC1937 is a BRCA1-defective breast cancer cell line which discloses higher sensitivity to cisplatinum (CDDP) as compared to the BRCA1 full-length cDNA transfected clone, HCC1937/wtBRCA1. By cDNA microarray whole gene expression profile and Ingenuity Pathway Analysis (IPA) software, we found a differential modulation of Notch signaling after CDDP exposure in HCC1937 cell line as compared to BRCA1-reconstituted clone with a strong down-regulation of Notch1, 2 and 3 expression together with other genes involved in the Notch signaling network, including Hes1 and Jag1. Notch pathway was not affected by CDDP exposure in BRCA1-reconstituted cells. By Western Blot analysis, we next analyzed if the different trascriptional response of the two cell lines to CDDP occurred together with down-regulation of Notch protein in BRCA1-defective cells. We found enhanced expression of BRCA1 in BRCA1-transfected cells. We then analyzed the Notch 3 protein's expression in HCC1937 and HCC1937/wtBRCA1. By Western blot analysis, we demonstrated a significant reduction of Notch 3 protein's expression following CDDP exposure at the IC50 dose for 12, 18 and 24 hours, in HCC1937 as compared to HCC1937/wtBRCA1, consistently with gene expression results. In the aim of evaluating the role of Notch signaling in the regulation of the pharmacological response to CDDP in BRCA1-defective cells, we exposed HCC1937 cells to the pan-Notch inhibitor, γ-secretase inhibitor XII (GSI-XII), alone or in combination with CDDP, and we elvaluated, by MTT assay, in vitro growth effects on cell survival. We observed a time and dose dependent decrease of cell growth of HCC1937 following GSI-XII exposure as compared to BRCA1-reconstituted clone, with an IC50 between 25 and 35 microM as single agent, while HCC1937/wtBRCA1 where highly resistant to the drug. The combination of GSI-XII plus CDDP produced a significant sinergistic antitumor effect at 48 hours in HCC1937 which did not occurred in HCC1937/wtBRCA1. Our findings suggest that the high sensitivity of BRCA1-defective cells to CDDP exposure may be related not only to depression of the DNA-damage repair machinery but also to down-modulation of the Notch survival pathway. Moreover, our data indicate that the antitumor activity of CDDP is enhanced by combination with the Notch inhibitor GSI-XII suggesting that γ-secretase inhibitors may represent a potential and novel therapeutic tool in this specific setting.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3128.
Examining the Role of ErbB2 in a Mouse Model of Type I Insulin like Growth Factor Receptor-Induced Mammary Tumourigenesis.
Authors: Campbell C. , Moorehead R.
Published: December 2010

Background: The type I insulin-like growth factor receptor (IGF-IR) and epidermal growth factor receptor 2 (ErbB2) are both receptor tyrosine kinases extensively implicated in human breast cancer. The IGF-IR is involved in cell proliferation and survival and is overexpressed in a large proportion of breast cancers. Similarly, ErbB2 (Her2) has been identified as a key contributor to a relatively large subset of aggressive breast cancer and is currently the basis for clinical therapeutics. To examine how the IGF-IR contributes to breast cancer and to study this receptor as a potential therapeutic target, we have previously created an inducible transgenic mouse model of IGF-IR overexpression in the mammary gland. In this model, tumours rapidly form upon induction of the transgene, with metastasis to the lung occurring in approximately 25% of animals. In addition, the RM11A cell line was isolated from tumour tissue and was shown to maintain inducible IGF-IR overexpression. The purpose of this study was to examine the interaction between IGF-IR and ErbB2 using our model.Materials and Methods: Protein expression was studied through Western blotting. ErbB2 expression was manipulated through the use of siRNA (for downregulation) and through the use of a plasmid encoding ErbB2 (for upregulation). Stable overexpression of ErbB2 was achieved through transfection of this plasmid with subsequent selection of stable integrants. Tumourigenicity was assessed by direct injection of these cells into the mammary gland. MTT assays and Ki67 staining (through immunofluoresence or immunohistochemistry) were used to examine cell survival and proliferation.Reults: In RM11A cells, survival was impaired upon downregulation and inhibition of ErbB2 in vitro. In our transgenic mouse model, ErbB2 was upregulated in primary tumours overexpressing the IGF-IR and was downregulated upon removal of IGF-IR induction and also in IGF-IR-independent recurrent tumours. In vivo, overexpression of ErbB2 conferred a more aggressive phenotype to RM11A cells. However, ErbB2 overexpression did not enhance proliferation in these cells in vitro or in vivo. The mechanism through which ErbB2 enhances tumourigenesis in or model is currently being investigated.Discussion: ErbB2 appears to have a role in IGF-IR-induced mammary tumourigenesis and can enhance tumour growth. Also, the IGF-IR appears to function in the regulation of ErbB2 in our model. Understanding how these two potent oncogenes interact to promote mammary tumourigenesis will ultimately help in designing treatment regimes specifically targeting the IGF-IR and/or ErbB2.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3156.
Identification of Interferon Regulatory Factor (IRF)-5 Tumor Suppressor Function in Human Breast Cancer.
Authors: Bi X. , Barnes B.
Published: December 2010

Background: Transcription factors of the IRF family participate in the early host response to pathogens, hematopoietic differentiation and immunomodulation. IRFs can regulate cell growth and apoptosis thereby affecting the susceptibility to and progression of cancer. IRF-1 was recently identified as a tumor suppressor in breast cancer. Early evidence from our lab and others suggests that IRF-5 may also have tumor suppressor activities. Although loss of IRF-5 expression has recently been shown to enhance transformation by Ha-ras, very little is known of IRF-5 tumor suppressor function(s).Material and Methods: Using paraffin-embedded formalin-fixed human breast tissue specimens from 64 patients with varying stages of ductal carcinoma, we analyzed and compared the expression of IRF-1 and IRF-5 by immunofluorescence (IF). Clinical data, such as histological subtype, stage, and receptor expression (ER, PR Her-2) were available for clinicopathological correlations. Protein levels were also determined in normal human breast epithelial MCF-10A, and -12A cells, and in cancer cell lines MCF-7, MDA-MB-231, -468, and T47D by immunoblot and IF. By retroviral infection, we overexpressed IRF-5 in breast cancer cell lines or knocked-down IRF-5 in normal MCF-10A and -12A cells and examined cell growth, apoptosis, metastasis and sensitivity to cytotoxic stress. In vivo tumor suppressor function was determined in NCr nu/nu mice inoculated with MCF-7 or MDA-MB-231 cells expressing or lacking IRF-5.Results: We confirmed recent data on IRF-1 revealing that it is expressed less frequently in tumor tissue of high-grade ductal carcinoma in situ (DCIS) or invasive DC compared with normal breast tissue. Data from patient samples indicate a significant difference between IRF-1 and IRF-5 expression that was distinctly associated with breast cancer disease stages, including early-to-late stage DCIS, suggesting their utilization as prognostic indicators.Similar findings were made in breast cancer cell lines. We demonstrate that forced expression of IRF-5 in breast cancer cells inhibits colony formation, sensitizes cells to cytotoxic stress-induced apoptosis, inhibits metastasis, and tumor formation in nude mice. Conversely, knockdown of IRF-5 in MCF-10A and -12A cells resulted in resistance to stress-induced cell death.Discussion: Data from this study strongly support a tumor suppressor function for IRF-5 in breast cancer. Ultimately, the study is expected to lead us to the identification of new prognostic indicators and therapeutic approaches for the treatment of breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3160.
p53 Mutation in Inflammatory Breast Cancer Cell Lines.
Authors: Li C. , Lee B. , Woodward W. , Ueno N. , Robertson F. , Reuben J. , Cristofanilli M.
Published: December 2010

Background: Inflammatory breast cancer (IBC) is the most aggressive manifestation of primary breast cancer and represents 1% to 2% of primary breast cancer in the United States. IBC is characterized by an acute inflammation of the skin of the affected breast generally believed to be caused by blockage of the dermal lymphatics by tumor emboli. Wild type (WT) p53 is a tumor suppressor gene, which induces apoptosis and p53 mutations are associated with poor prognosis in breast cancer. Compared with locally advanced breast cancers, IBC patients have higher levels of mutated p53 protein that has been associated with more aggressive tumors, anthracycline resistance, shorter progression free survival, shorter overall survival, and less favorable long-term outcome. The aim of this study was to determine if there are unique genetic variations in IBC cell lines that would provide specific genetic p53 mutations that could be exploited for targeted therapy with the intent of improving response to treatment and overall survival in IBC.Materials and Methods: Genomic DNA was extracted from six breast cancer cell lines (MDA-453, SUM149, MCF-7, KPL4, MDA321, and SUM190) and the immortalized human mammary epithelial cells (HMLE) using the Qiagen DNA Blood Mini Kit (Valencia, CA). Among the 6 breast cancer cell lines, KPL-4, SUM149 and SUM190 are IBC cell lines; MDA231 and SUM149 have basal-like phenotype; MCF-7 has wild-type p53; SUM190 and KPL-4 are Her2 amplified. The DNA purity and concentration were determined by spectrophotometric measurements of absorbance at 260nm and 280 nm. Polymerase chainreaction (PCR) was performed to amplify the fragments of exons 2-11 of the p53gene using consensus primers. The PCR products were scanned and identified using the Agilent Bioanalyzer 2100. DNA sequencing was performed on PCR products in the ABI PRISM 310 Genetic Analyzer. The BLAST search was used to identify p53 mutations compared with the reference sequence, X54156, from Genbank.Results: We screened 2-11 exon sequences of the p53 gene in the 7 human breast cell lines. We identified two IBC cell lines (SUM149, SUM-190) with a p53 gene alteration that predicted a change in the encoded protein, SUM149 at exon 7 (ATG to ATA, Met-237-Ile) and SUM190 at exon 9 (CAG to TAG, Gln -317-stop). Both mutations have been previously reported. Five nonsense mutations were identified in two other cell lines, MDA-453 and MDA321. No mutations were identified in KPL4 and HMLE cells.Conclusions: The p53 mutation profile in breast cancer cell lines suggests an additional biological feature for the characterization of IBC. Furthermore, these data support the previously reported association between p53 status and chemo- and radioresistance in this disease responsible for poor prognosis. Therapies directed to restore p53 function should be explored in IBC models and in clinical trials.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3161.