bluuebunny/crossref_metadata_2025_split huggingface dataset:
P2-01-26: TIMPv: a novel downstream transcriptional target gene that underpins CD146-suppressed breast tumor invasion
Authors: Ouhtit A , Abdelmageed Z , Fernando A , Gaur RL , Raj MHG 
Published: June 2012

Abstract
Background: CD146, a marker of endothelial cells, promotes tumor progression of many cancers including melanoma and prostate. Strikingly, several lines of evidence suggest that it is a suppressor of breast cancer (BC) progression. Not only the ligand(s) has not been identified, but CD146-downstream signaling mechanisms remain unknown.
Material and Methods: Here, we report the functional importance of CD146 and that of a novel transcriptional target of CD146-signaling, variant of tissue inhibitor of metalloproteinases (TIMPv) identified by microarray analysis, in underpinning the suppression of BC invasion, using novel validated Enhanced Green Fluorescent Protein (EGFP)-inducible systems of CD146 expression both in vitro and in vivo. Results: In functional experiments, induction of CD146 inhibited BC cell migration and invasion. TIMPv was identified by expression profiling as a novel transcriptional target of CD146-signaling, an association validated by quantitative PCR and immunoblotting experiments in a range of breast and melanoma cancer cells. However, siRNA inhibition of CD146 in the SKMel-28 melanoma cell line increased TIMPv expression, suggesting that while TIMPv is a positive transcriptional target of CD146 in BC cells, it is negatively regulated in melanoma cells. Furthermore, the functional relevance of TIMPv to CD146-suppressed metastasis was demonstrated by selective suppression of TIMPv in CD146-expressing BC inducible cells using RNAi. More interestingly, induction of CD146 expression in vivo using mouse EGFP-inducible system of CD146 expression resulted in suppression of breast tumor growth.
Discussion: Our study is the first report to provide a functional molecular link of TIMPv to cancer via unique axis that underpin CD146-suppressed BC progression, thus identifying TIMPv as a potential target for guiding the development of novel therapeutic strategies for BC.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-26.
P2-05-01: Acquired Tamoxifen Resistance Promotes Angiogenic Responses in ER+ Breast Cancer Cells.
Authors: Hayes E , Smith C , Nicholson RI , Gee J , Hiscox S 
Published: June 2012

Abstract
Background: In the treatment of pre-menopausal women with oestrogen positive (ER+) breast cancer, tamoxifen represents a first line of adjuvant treatment with demonstrable benefits. Despite this, resistance is frequently acquired to tamoxifen with an associated poor prognosis. Breast cancer cell models have revealed the importance of growth factor signalling networks in sustaining growth of endocrine-resistant cancers and, more recently, their ability to promote a highly migratory and invasive phenotype, together with the expression of genes with pro-angiogenic ontology. The potential of endocrine resistant cells to elicit angiogenic responses, however, remains unknown.
Materials and Methods: Real-time PCR was used to validate results from preliminary Affymetrix-based gene profiling of pro-angiogenic gene expression in endocrine-sensitive MCF7 cells and their tamoxifen-resistant (TamR) counterparts. The expression of pro-angiogenic factors in conditioned media (CM) from these cells was assessed by ELISA. The proliferative and migratory effects of MCF7 and TamR CM on vascular endothelial cells (HUVEC and HECV cells), was determined by MTS cell proliferation assay, wound closure assays and Matrigel tubule formation assays. Changes in endothelial cell migration following co-culture with endocrine-resistant cells were examined using Boyden-chamber chemotaxis assays. Growth factor signalling and migration pathway activation in endothelial cells in response to CM was determined by Western blotting. Results: TamR cells were found to express high levels of HIF-1α, IL-8 and VEGF-A at an mRNA level compared with expression in MCF7 cells. High levels of VEGF-A protein were also confirmed in the conditioned media from TamR cells versus their endocrine-sensitive counterparts. TamR conditioned media promoted endothelial cell proliferation, migration and the formation of tubules to a greater extent than that seen in MCF7 CM treated cells. TamR conditioned media was found to stimulate VEGFR2 phosphorylation and downstream activation of MAPK in endothelial cells compared to MCF7 CM. Pharmacological inhibition of VEGFR2 activity in endothelial cells suppressed TamR-induced endothelial cell proliferation and VEGFR phosphorylation. Further pharmacological manipulation of erbB receptors and intracellular kinases in TamR cells revealed an ERGF/Her2-Src kinase dependent mechanism of VEGF-A production in these cells.
Discussion: These data suggest acquired tamoxifen resistance is accompanied by development of an erbB receptor/Src kinase-dependant pro-angiogenic phenotype which, if recapitulated in vivo, may promote tumour progression. Therapeutic targeting of erbB/Src axis may prove beneficial in such cases.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-01.
P2-05-08: Expression of VE-Statin/egfl7 in Breast Cancer.
Authors: Lauridant-Philippin G , Baranzelli M-C , Fournier C , Samson C , Soncin F , Bonneterre J 
Published: June 2012

Abstract
Patients and methods: Specimens of breast cancer (174 invasive and 37 in situ) were obtained from 205 patients (pts) treated in our institution in 2005.Their median age was 57 years (39-92). VE-statin expression was studied by immunohistochemistry (IHC) using anti-hVE-statin/egfl7 (AF3638, R&D) and by RT-qPCR on formalin-fixed paraffin sections. 174 invasive carcinomas (141 ductular and 30 lobular) and 37 DCIS were studied. Determination was done twice by two different observers. Statistics used Chi2 and Fisher's exact tests for categorical data and Kruskall-Wallis test for continuous variables.
Results: Pathological characteristics of the patients with invasive tumors were as follows: 81% ductular, 17% lobular; median tumor size 18 mm (2-100); SBR grade 1 in 27%, 2 in 58% and 3 in 15% of the tumors; estradiol receptor (ER) + in 90%, progesterone receptor (PR) + in 66%; 6,5% HER2 positive. In 105 cases there was DCIS together with invasive tumors. For invasive tumors and DCIS, progression-free survival (PFS) at 5 years were 85% and 100% and overall survival (OS) 96% and 95%, respectively. Characteristics of the DCIS were median tumor size 18 mm (2-60), ER+ in 88%, PR+ in 62%. IHC results were obtained in 159 invasive carcinoma and 34 DCIS, RT-qPCR results in 96 samples. In the group of invasive tumors, a cytoplasmic VE-statin/egfl7 signal was obtained in 80% of invasive cells, more frequently than in normal glandular cells (71% vs 41%, p=0,003) but significantly less than in peritumoral DCIS (71% vs 89%, p=0,001). The signal was higher in ductular than in lobular carcinoma (85% vs 54%, p=0,003). In RT-qPCR analyses, RNA levels were higher in SBR1 grade tumors (p=0,008), in ER+ invasive tumors (p=0,02) and in pN- tumors (p=0,003). In the group of DCIS, a cytoplasmic signal was obtained in 82% of the cells. It was more frequently observed in tumor than in normal glandular cells (82% vs 52%, p=0,02). A 3+ cytoplasmic signal was more frequent in DCIS alone than in DCIS associated with invasive carcinoma (23% vs 6%, p=0,02). No prognostic significance differences were observed on PFS or on OS.
Discussion: This work is the first expression study of VE-statin/egfl7 in breast cancer. The results were reproduced by two different observers. IHC signal was cytoplasmic and higher in DCIS than in invasive carcinomas. Furthermore the signal was even higher when there was only DCIS than when it was associated with invasive lesion. The signal was higher in ductular than lobular carcinoma, consistent with previous publications on other angiogenesis mediators. Similarly, the signal was higher in low SBR grade, as well in ER positive tumors. Phase 1 studies are ongoing in order to evaluate the effects of inhibition of VE-statin/egfl7.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-08.
P2-07-01: Copy Number Variants in Early-Onset-Breast Cancer.
Authors: Ludwig S , Ivanovich J , Graubert TA , Goodfellow PJ 
Published: June 2012

Abstract
Background: Copy number variants (CNVs) are known to contribute to the risk for development of immune diseases, neurological disorders and to a lesser extent cancer. Many CNVs do not directly involve gene sequences. CHL1, a cell adhesion molecule with homology to L1CAM, is down-regulated in a large proportion of breast cancer. A deletion CNV immediately 5’ of the CHL1 transcriptional start site has been described. The CHL1 CNV's role in breast cancer risk has not previously been explored.
Material and Methods: 50 research subjects recruited to the Washington University/Siteman Cancer Center Young Women's Breast Cancer Program (YWBCP) were selected for a CNV discovery study. These women were diagnosed breast cancer &lt; age 40, tested negative for BRCA1/2 mutations and have a negative family history for breast cancer. The selection criteria enriched for de novo mutations. Comparative genomic hybridization (CGH) analyses were undertaken using the NimbleGen 2.1 million oligonucleotide array. DNA copy number was compared between proband and each of her parents to identify potential deletions or amplifications in the daughter relative to both parents (de novo changes). PCR was used to confirm a subset of the CGH-predicted differences in copy number. Chi-square test was used to determine statistical significances of CNV occurrences near the CHL1 gene between women with breast cancer and control cases.
Results: aCGH data analysis has been completed for 25 YWBC trios (patients and parents). 247 differences in copy number in the breast cancer cases relative to their parents were identified (1-18 putative de novo CNVs per case). 91% of the observed changes were deletions and 9% were amplifications. One proband showed an 11,560 bp deletion 5’ of the CHL1 gene that was not seen in her parents. Conventional PCR confirmed a homozygous deletion of a ∼3 kb region in the patient, in the region of a known CNV with a reported frequency of ∼3%. Testing an additional 368 early-onset breast cancer cases for the CHL1 deletion revealed 25 additional homozygous deletions (6.8%). The deletion was present in 9 of 197 controls (4.5%). qPCR studies revealed the parents of the patient with the CHL1 deletion each carried a single copy of the CNV, making de novo copy number change in the proband very unlikely. Deletion specific assays for the CHL1 CNV are being used to test CNV genotypes in an extended case-control study.
Discussion: CHL1 expression has previously been shown to be down-regulated in primary breast tumors. Our preliminary analysis of YWBC cases and controls indicates the CHL1 regulatory region CNV is unlikely to be associated with risk for early-onset breast cancer. Additional candidate de novo copy number changes seen in the genomes of patients with early-onset breast cancer are being evaluated to determine their roles as genetic risk factors.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-07-01.
P2-08-08: Benefit of Preoperative Breast MRI in Patients with Newly Diagnosed Breast Cancer.
Authors: Miesbauer M , Sheu H , Schoissengeier A , Kirchweger B , Waldenberger P 
Published: June 2012

Abstract
Objective: The aim of our study was to evaluate the effect of routinely performed preoperative Breast MRI (pMRI) on the planned therapeutical management of all patients with newly diagnosed Breast cancer.
Subjects and Methods: 180 patients with newly diagnosed Breast Cancer between 9/2009 - 12/2010 were enrolled in this retrospective study performed in a certified European Breast Cancer Unit; mean age 60,6 years; 1,5 T MRI ( TIM-Symphony, Siemens Germany).
Results: In 53/180 (29,4%) cases the preoperative MRI changed the planned therapeutical management concerning surgical and chemotherapeutical options. In 42/180(23,3%) patients additional lesions were identified only by pMRI, 33/42(78,6%) have been malignant and 8/42(19%) were benign. The real extent of the tumor was detected in 43/53 (81%) cases — group A; a bilateral carcinoma was detected in 4/53 (7,5%) cases — group B; in 6/53 (11,5%) cases the malignant lesion could only be detected with preoperative MRI — group C; for group A the pMRI led to wider excisions with 15 segmental -or quadrant resections and 28 mastectomies; for group B the pMRI resulted in 4 additional lumpectomies; for group C the pMRI caused 4 lumpectomies, 1 segemental resection and 2 mastecomies .5 of 6 patients got neoadjuvant chemotherapy only due to results of pMRI; in 3 of these 5 patiens breast conserving surgery was possible. Histology: DCIS 23/53 (43,4%), invasive cancer 23/53 (43,4%), multicentric/multifocal malignant lesions 7/53 (13,2%). The sensitivity of the pMRI was not influenced by the histology of the lesion, the age of the patient or menopausal status. The maximum periode between initial presentation of the patient and breast surgery was 21 days.
Conclusion: In 29,4% of our patients the pMRI resulted in a significant change of the planned therapeutical regime. Histology of the lesion, patients age or menopausal status did not influence the sensitivity of pMRI. There was no relevant delay in the initiation of treatment. Therefore we recommend performing pMRI in breast cancer staging routinely.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-08-08.
P2-09-14: Evaluation of Angiogenesis Using Synchrotron Radiation in Xenograft Mouse Models of Breast Cancer.
Authors: Gu S-M , Zhang X , Li R-M , Jin W , Shen Z-Z , Shao Z-M , Xu X-M , Wu J 
Published: June 2012

Abstract
Visualization of microvasculatures is an important step in understanding the mechanism of early vessel disorders in breast cancer and developing effective therapeutic strategies. However, quantitative evaluation of breast cancer angiogenesis using immunohistochemistry has been limited in generating detailed and reproducible data. To analyze the diversification of angiogenesis in the development of tumor growth and evaluate anti-vascular effects of Avastin, a new method of microvascular imaging was introduced in this paper. We used New X-ray microangiography and third-generation synchrotron radiation-based micro-computed tomography (SR micro-CT) to investigate the structures and density of microvessels at each stage of xenograft mouse models that were created by inoculation with MDA-MB-231HM cells (one breast cancer cell line with high metastasis potential). Barium Sulfate Nanoparticles, was used as a blood vessels contrast agent through left cardiac ventricle. Three dimensional structures of microvessels were displayed with a high spatial image resolution at around 20–30 μm. The density of microvessles was significantly reduced from the third week in mouse xenograft models. This observation with SR micro-CT was consistent with the results analyzed by immunohistochemical techniques. In addition, human breast cancer-bearing mice were treated site-specifically with humanized monoclonal antibody (Avastin) that targets all isoforms of VEGF-A. Tumor volume and the density of angiogenesis were significantly reduced in xenograft mouse models of breast cancer treated with avastin when compared with untreated mice as assessed by SR micro-CT. Specifically, the density of smaller vessels (di&lt;50μm) was decreased significantly while the density of larger vessels was little changed in the Avastin-treated mice. Therefore, synchrotron radiation technique is a novel method for investigating breast cancer angiogenesis, and this might be considered as an additional complementary tool for more precise quantification of angiogenesis.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-09-14.
P2-11-03: High Expression of CX3CL1 by Tumor Cells Correlates with a Good Prognosis and Increased Infiltrating CD8+ T Cells, Natural Killer Cells, and Dendritic Cells in Breast Carcinoma.
Authors: Lee JS , Park MH , Yoon JH 
Published: June 2012

Abstract
CX3CL1 is the only CX3C chemokine that can chemoattract CD8+ T cells, natural killer (NK) cells, and dendritic cells (DCs). Although studies have reported that CX3CL1 regulates the host immune response, the roles of the CX3CL1 in breast carcinoma remain unknown. We evaluated the expression CX3CL1 in 198 surgical specimens of breast carcinoma and analyzed any association with the clinicopathological factors, including CD8+ T cells, CD56+ NK cells, and CD1a+ DCs, while also evaluating the impact on the prognosis. High CX3CL1 expression was found in 136 of 198 (68.7%) breast carcinomas. The numbers of stromal CD8+ T cells, intratumoral CD1a+ DCs, and stromal CD56+ NK cells were significantly increased in the high CX3CL1 expression group (19.3 ± 15.2, 0.3 ± 0.9 and 1.4 ± 1.7 per field, respectively) compared with the low expression group (10.2 ± 13.1, 0.0 ± 0.3 and 0.7 ± 0.8 per field, respectively). Patients with high CX3CL1 expression had a significantly better prognosis for disease-free and overall survival than those with low expression (P = 0.002 and P = 0.000, respectively). In a multivariate analysis, the CX3CL1 expression was identified as one of the independent prognostic factors for overall survival (odds ratio, 2.29; P = 0.002). These data suggest that CX3CL1 expressed in the tumor cells appears to enhance the recruitment of CD8+ T cells, NK cells, and CD1a+ DCs, thereby bringing about a better prognosis in breast carcinoma. CX3CL1 is a new prognostic biomarker and be a novel candidate for development of a more effective therapeutic strategy for breast carcinoma.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-11-03.
P2-11-11: IGFBP Ratio Confers Resistance to IGF Targeting and Correlates with Increased Invasion and Poor Outcome in Breast Tumors.
Authors: Becker MA , Hou X , Harrington SC , Carboni JM , Gottardis MM , Haluska P 
Published: June 2012

Abstract
Purpose: To assess the role of insulin-like growth factor binding protein 5 (IGFBP-5) as a marker of relapse and survival in breast cancer tumors.
Experimental Design: Targeted regulation of IGFBP-5 was identified in MCF-7 cells resistant (MCF-7R4) to the IGF-1R/InsR inhibitor, BMS-536924 and examined by comparative microarray analysis, western and ELISA. Protein expression of IGFBP-5 was measured by immunohistochemistry in a cohort of 84 breast cancer patients to examine correlative associations with invasive tumor fraction and overall survival (OS). The expression ratio of IGFBP-5/IGFBP-4 (BPR) was determined in multiple breast tumor cohorts for univariate analysis.
Results: IGFBP-5 was markedly upregulated and highly localized to the membrane in MCF-7R4 resistant cells. When compared to pathologically normal reduction mammoplasty tissue, IGFBP-5 expression levels were upregulated in both invasive and histologically normal adjacent breast cancer tissue. In an independent cohort of breast cancer patients, IGFBP-5 protein levels correlated directly with invasion and OS. In univariate and multivariate modeling, metastasis-free survival, recurrence free survival (RFS) and OS were significantly associated with high IGFBP-5 expression. Prognostic power of IGFBP-5 was further increased with the addition of IGFBP-4 and tumors were ranked based upon IGFBP-5/IGFBP-4 expression ratio (BPR). Multiple breast cancer cohorts confirm that BPR (high vs. low) was a strong predictor of RFS and OS.
Conclusion: IGFBP-5 expression is a marker of poor outcome in breast cancer patients. An IGFBP-5/IGFBP-4 expression ratio may serve as a surrogate biomarker of IGF pathway activation and predict sensitivity to IGF-1R-targeted therapies.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-11-11.
P2-12-02: Correlation between BMI and Clinical Outcome of Patients with Early Stage HER2+ Breast Cancer from the N9831 Clinical Trial.
Authors: Crozier JA , Moreno-Aspitia A , Ballman KV , Martino S , Kutteh LA , Davidson NE , Kaufman PA , Perez EA 
Published: June 2012

Abstract
Background
Obesity, as defined by body mass index (BMI), has been associated with increased recurrence rate, shorter DFS and increased death rates due to breast cancer (BC). Most of the studies to date have examined the relationship of BMI and DFS in patients with hormone receptor positive disease. To our knowledge, BMI and its relationship with outcome in early stage HER2 positive breast cancer has not previously been examined. The N9831 is a large phase III trial testing the role of trastuzumab in the adjuvant setting of high risk patients with early stage HER2+ BC. We hypothesized that the occurrence of overweight and obesity may correlate with outcome.
Methods: This analysis presents BMI and its relation to tumor characteristics and DFS in patients (pts) enrolled in the N9831 clinical trial. Pts were categorized as normal weight, overweight or obese using the WHO BMI classification parameters of &lt; 25%, 25–29% and ≥ 30% respectively. For patient characteristics, patients were grouped into non-obese (BMI&lt; 30) and obese (≥ 30) cohorts. DFS was estimated by the Kaplan-Meier method. Comparisons between arms A (chemotherapy alone), B (chemotherapy plus sequential trastuzumab) and C (chemotherapy plus concurrent trastuzumab) were performed using the Cox proportional hazards model, stratified by BMI.
Results: Analysis was completed on 3,017 eligible pts. Obese pts were more likely to be older and postmenopausal (p&lt;0.0001 for both). There was no significant association between BMI and ER/PR status (p=0.07) or histologic tumor grade (p=0.33). Obese pts were found to have significantly larger tumors ≥ 2 cm (p=0.002) and more positive lymph nodes (p=0.02). There was no significant difference in DFS within each intrinsic arm (A, B and C) between the obese and non-obese pts at 3, 5 or 7 yrs of follow up. However, pts in the non-obese group had significantly improved DFS in arm B and C compared to arm A (p=0.001 and p&lt;0.0001 respectively). Also obese pts in arm C had significantly improved DFS compared to obese pts in arm A (p=0.008). There was a trend of improved DFS in the obese group in arm B compared to arm A, but this was not statistically significant (p=0.09). Pts in the normal weight and overweight groups did significantly better in arm B (p=0.02 for both) and arm C (p=0.01 and p=0.002 respectively) compared to arm A.
Conclusions: This analysis of data from the N9831 study confirms that obese pts with early stage HER2+ tumors have worse clinical outcome than pts with BMI &lt; 30%. Adjuvant trastuzumab improved clinical outcome regardless of BMI. This study supports weight loss intervention for obese women with early stage HER2+ BC.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-02.