bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P4-06-18: Asporin Is Associated with Invasive Growth and Bone Metastasis of Breast Cancer
Authors: Bouchal J , Simkova D , Kharaishvili G. 
Published: April 2011

Abstract
Background: In an earlier publication we identified asporin as a novel cancer related protein (BMC Cancer 2007, 7:55), also reported among bone metastasis specific genes (Cancer Letters 2009, 276:212-220). Asporin is a member of the class I small leucine-rich repeat proteoglycan family and besides affinity to collagen, it binds calcium and initiates collagen mineralization. Asporin is abundantly expressed in tissues with high remodeling rate (e.g. peridontal ligament) whereas extremely high turnover of matrix proteins is also associated with invasive cancer. Material and methods: Formalin-fixed paraffin-embedded tissues [invasive ductal (101) and lobular (39) carcinomas classified according to hormone receptors and Her-2; 5 bone metastases from autopsies] were immunohistochemically stained with in-house polyclonal peptide antibody against asporin. The antibody was validated on uterus tissue as a positive control together with a commercial antibody (Abcam). Data were evaluated by non-parametric Mann-Whitney U test and Spearman correlation coefficient. Expression of asporin in both tissues and cell lines was also queried in databases Gene Expression Omnibus, ArrayExpress and Oncomine. Expression of asporin was further tested by RT-PCR in selected breast cancer cell lines. Migration and invasion of MDA-MB-231 cells were tested using real-time cell analysis Xcelligence system (Roche). Transwells were coated by solidifying collagen for 6 hours (with or without recombinant asporin). Starved cells were seeded on the collagen in transwells while FBS was used as a chemoattractant in bottom wells. Results: Levels of asporin were significantly higher in invasive lobularcarcinomas than ductal ones (P&lt;0.001). Asporin was also highly expressed in all bone metastases. Asporin expression positively correlated with tumor grade within the triple negative subgroup (Rs 0.4, P&lt;0.05) but not in luminal and Her-2 subtypes. Expression of asporin in invasive breast carcinomas was also confirmed in other public datasets (e.g. Karnoub breast and Hess breast in Oncomine database). Surprisingly, we found no breast cancer cell line overexpressing asporin, which underlines the importance of the tumor microenvironment in vivo. MDA-MB-231 cells migrated faster through collagen I with recombinant asporin.
Conclusion: Asporin is expressed by both invasive breast carcinomas and bone metastases. Mechanistically, asporin may affect extracellular matrix structure and promote invasive growth of breast cancer cells. Further investigation of the role of asporin in metastasizing to bone is additionally justified given its affinity for calcium. Acknowledgements
This work was supported by grants NS 9956-4 from the Czech Ministry of Health and MSM 6198959216 from the Czech Ministry of Education.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-06-18.
Abstract P4-06-22: Persistent Triple-Negative Phenotype Is Associated with Poorest Outcome among Patients with Metastatic Breast Cancer (BC) - Results of the Retrospective Multicenter PriMet Study Comparing Molecular BC Phenotypes in Primary Tumors and Corresponding Recurrences
Authors: Liedtke C , Gluz O , Heitz F , Freudenberger M , Würstlein R , Hungermann D , Ortmann M , Kates RE , Nitz U , du Bois A , Fehm TN , Harbeck N. 
Published: April 2011

Abstract
Background: Patients with triple negative (TN) breast cancer (BC), defined by lack of both ER/PR expression and HER2 overexpression, have an unfavorable prognosis. Although phenotype changes between primary tumor (PT) and disease recurrence (DR) have been described, their clinical significance is still unclear.
Methods: We conducted PriMet, a retrospective multicenter (n=11) study to compare BC phenotype in PT and corresponding DR. Inclusion criteria comprised (1) unilateral BC with subsequent/synchronous local/regional/distant DR and (2) immunohistochemical confirmation of DR. Our aim was to (a) evaluate discordance rates between PT/DR, (b) find predictors for discordance, and (c) analyze the impact of discordance on patient outcome.
Results: 436 patients were entered into PriMet; 414 had no evidence of primary metastatic disease (M0). Median follow-up in patients alive at time of analysis was 73. 1 (4.4-293.6) months. Triple receptor status for PT and DR was available in 377 patients; 68 patients (18.0 %) showed TNBC in PT, 40 patients (10.6 %) had TNBC in both PT and DR (i.e. TNBC persistence); 28 patients (7.4 %) changed from TNBC to non-TNBC (15 became HER2 positive (4.0 %)). Status changes for ER and PR were significantly positively associated (P&lt;0.001). Patients with either persistent TNBC or non-TNBC had a median age at diagnosis of 52.0 compared to 55.5 yrs for patients with discordant TN status (p=0.04). Compared to differing constellations, persistent TNBC was associated with higher tumor grade (p=0.018) as well as with both decreased disease-free survival (DFS) (see figure) and post-recurrence survival (PRS) in M0 patients. In multivariate analysis containing pT stage, nodal stage, tumor grade in PT, TNBC in PT and TNBC persistence, TNBC persistence remained significantly associated with decreased DFS and PRS.
Conclusion: PriMet provides substantial evidence regarding potential phenotype changes in PT vs. DR and underlines the importance of immunohistochemical DR verification even in initially TN disease. TNBC persistence was significantly associated with adverse patient outcome.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-06-22.
Abstract P4-07-02: Expression of Cell Adhesion Molecules Predicts Prognosis in Early Breast Cancer Patients
Authors: de Kruijf EM , Saadatman S , Sajet A , Boer W , van Velzen N , Putter H , Smit VTHBM , Liefers GJ , van de Velde CJH , Kuppen PJK. 
Published: April 2011

Abstract
Purpose: New prognostic and predictive factors are sought for improvement of tailored treatment in early breast cancer. We examined the clinical impact of cell adhesion molecules (CAM): E-cadherin, N-cadherin, Ep-CAM and CEA.
Patients and Methods: Our study population (n=574) consisted of all early breast cancer patients primarily treated with surgery in our center between 1985 and 1994. A tissue micro array (TMA) of formalin-fixed paraffin-embedded tumor tissue was immunohistochemically stained for expression of mentioned CAM. The percentage of membranous stained cells was microscopically analyzed. Based on the median score, all CAM were classified in two groups: low expression versus high expression. For CEA, high expression was further subdivided based on the intensity of staining: high expression and highest expression. Results: High expression was seen for E-cadherin, N-cadherin and Ep-CAM in 49%, 46%, 27% of patients respectively. Low expression, high expression and highest expression were found in respectively 48%, 45% and 8% of cases for CEA. Low expression of E-cadherin (p=0.015) and higher expression levels of N-cadherin, Ep-CAM, CEA (p=0.004; 0.046; 0.001 respectively) all resulted in a worse relapse free period (RFP) of patients. Multivariate analysis revealed only E-cadherin and CEA to be independent prognostic variables. A combination variable was created with expression of both markers: (1) E-cadherin high expression, (2) E-cadherin low or CEA low or high expression (3) CEA highest expression. This variable revealed to be an independent prognostic parameter with high discriminative power for RFP (P&lt;0.001, E-cadherin low or CEA low or high expression versus E-cadherin high expression: Hazard Ratio (HR)= 1.9; CEA highest expression versus E-cadherin high expression: HR= 3.6). A statistically significant interaction was found between expression of both CAM (P&lt;0.001), suggesting a biological connection in their functioning.
Conclusion: We have demonstrated that E-cadherin, N-cadherin, Ep-CAM and CEA are of prognostic influence on outcome concerning RFP in breast cancer patients. A combined variable of E-cadherin and CEA expression revealed to have prognostic influence on RFP with high discriminative power and therefore is a candidate parameter for future outcome prediction of patients.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-07-02.
Abstract P4-07-06: Yes-Associated Protein, a Proposed Tumour Suppressor, Is Inversely Correlated to Cyclin D1 in Breast Tumours and Associated with a Worse Prognosis
Authors: Lehn S , Tobin NP , Sims AH , Jirström K , Landberg G. 
Published: April 2011

Abstract
Background: In breast cancer, cyclin D1 over-expression and CCND1 (11q13) gene amplification have been linked to a worse prognosis. Conversely, lower levels of cyclin D1 have also been correlated to an unfavourable outcome. In a microarray analysis of cyclin D1 silenced MDA-MB-231 cells we identified Yes-associated protein (YAP), located at 11q22, as one of the most down-regulated genes. As such, we aimed to determine the importance of YAP in relation to cyclin D1 in breast cancer biology.
Material and methods: Validation of cyclin D1, YAP antibodies and protein levels were performed in MDA-MB-231, MDA-MB-468 and MCF-7 by western blot. YAP protein expression was examined by screening 144 breast tumours and gene expression assessed by meta-analysis of six previously published breast cancer datasets totalling 1107 tumours with clinical follow-up. Subsequent data analysis was performed in SPSS. Results: YAP protein levels were down-regulated in cyclin D1 silenced MDA-MB-231, MDA-MB-468 and MCF-7 cell lines as predicted by microarray analysis. Surprisingly, YAP and cyclin D1 were inversely correlated at both protein and gene level in primary tumours (p=0.004 and P&lt;0.001). Amplification of the CCND1 gene was correlated to low YAP protein expression (P&lt;0.001) in the screening tumour material. Low YAP mRNA was an independent prognostic factor of recurrence (HR= 1.30, 95% CI 1.009-1.678, p=0.043) in the gene expression dataset. This effect was particularly pronounced in the luminal A subtype of breast tumours. Discussion: The independent prognostic value of YAP strengthens the notion of this protein as a tumour suppressor in breast cancer. The regulation of YAP by cyclin D1 appears to occur on both the protein level as seen in cell lines, and on the chromosomal level in terms of 11q aberrations. The inverse correlation observed in the tumour materials between cyclin D1 and YAP may be partly explained by chromosomal rearrangements; the amplification of the CCND1 gene (11q13) is often accompanied by loss of the 11q22 region of the chromosome, harbouring the YAP locus. Further studies are ongoing to clarify the link between YAP and cyclin D1 in breast cancer biology.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-07-06.
Abstract P4-07-08: High Level of Nuclear S100P Is Associated with Poor Survival in Early Breast Cancer Patients
Authors: Maciejczyk A , Lacko A , Ekiert M , Halon A , Matkowski R , Surowiak P. 
Published: April 2011

Abstract
Background: S100P — low molecule acidic protein, which belong to the EF-hand superfamily of calcium binding proteins has been shown to be involved in processes of proliferation, survival, angiognesis and metastasis in various human malignancies. In breast cancer S100P expression is associated with immortalization of neoplastic cells, progression and aggressive tumor behaviour indicating that this protein may have prognostic value. Several studies also revealed relationship between S100P level and resistance to cytotoxic drugs, however these observations are conflicting. Materials and methods: We analyzed nuclear and cytoplasmic expression of S100P in 85 stage II breast cancer (BC) patients with 17 years follow up. Immunohistochemical (IHC) reactions were performed on paraffin sections of primary tumours, using monoclonal antibodies against S100P. Semi-quantitative scale of ImmunoReactive Score (IRS) was applied (score 1-12) to evaluate the S100P expression level. Results were correlated with clinical and pathologic data. The Kaplan-Meier method was used to estimate the survival probabilities.
Results: IHC staining for S100P was predominantly nuclear (S100Pn). Of the 85 specimens evaluated, 23 (27,0%) were not stained, 35 (41,2%) exhibit borderline or weak expression (IRS: 1-4) and 27 (31,8%) strong expression (IRS: 6-12). Patients with strong S100Pn expression had significantly lower overall survival (OS) (P&lt;0,00001) and disease free survival (DFS) (p = 0,00002). Cytoplasmic S100P (S100Pc) expression did not appear to impact prognosis. When stratifying for lymph node status, S100Pn was a significant prognostic factor only in node-positive (N/+/) group and had borderline significance in N/-/(p =0,05777 and p=0,04105, for DFS and OS respectively). In contrast to other studies we did not find statistically significant correlation between S100P expression and tumor variables such as: lymph node involvement, estrogen receptor (ER), progesterone receptor (PR) and HER-2 status. Discussion (albo Conclusion): We demonstrated that high S100Pn expression level is associated with poor survival in early stage BC patients. Since preliminary data indicate that expression of S100P is up-regulated by activation of glucocorticoid receptor and other agents have potential to activate or inhibit S100P promoter activity, this protein may become a therapy target and warrants further studies with respect to its prognostic, predictive and potentially therapeutic value. Key words: breast cancer, prognostic biomarker, S100P
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-07-08.
Abstract P4-08-02: A Comparison between AQUA Quantitative Fluorescent Immunohistochemistry and Conventional Immunohistochemistry for Hormone Receptors
Authors: Bartlett JMS , Gustavson M , Stocken D , Rimm D , Christiansen J , van de Velde CJH , Hasenburg A , Kieback D , Putter H , Brookes C , Markopoulos C , Dirix L , Robson T , Seynaeve C , Dolled-Filhart M , Jones C , Graves L , McGuire J , Rea D. 
Published: April 2011

Abstract
Background: We have previous data showing that quantitation of hormone receptors can be highly informative in determining risk of early relapse in ER positive early breast cancer treated with tamoxifen or exemestane. Both quantitative immunohistochemistry (QIHC) and flouresecent immunohistochemistry (F-IHC as measured by AQUA technology) are highly prognostic over a wide expression range. We have explored the results of both assays to determine if current assays provide maximum information using current approaches.
Patients &
Methods: Pathology blocks from 4598 TEAM patients were collected and tissue microarrays constructed. Quantitative AQUA and IHC analysis (using image quantitation) of ER and PgR was performed centrally (Edinburgh & HistoRx). Results from both assays were compared and their prognostic impact on DFS at 2.75 years examined.
Results: Both AQUA and QIHC demonstrated linear relationships between intensity of staining for either ER or PgR and DFS at 2.75 years. For both PgR and ER AQUA provided significantly greater prognostic information that QIHC. However AQUA staining explained only 29% and 68% of the variability in ER and PgR QIHC results by logistic regression. Using both AQUA and QIHC data in a forward stepwise selection survival model demonstrated that AQUA and QIHC provided similar prognostic information over 70% and 50% of the range for ER and PgR respectively. High ER QIHC and low ER AQUA scores, and low PgR IHC and high PgR AQUA scores provided prognostic information unique to either platform.
Conclusion: Both QIHC and AQUA analysis of HR expression provides significant and highly important information on DFS risk in early breast cancer. It appears that these two platforms provide overlapping prognostic information and that the range of ER and PgR expression which impacts patient outcome is wider than measured by either system alone. Further investigation of the clinical significance of this broader range of hormone receptor expression in treatment decisions is warranted.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-08-02.
Abstract P4-08-06: Companion Diagnostic Test for Expression of β-III tubulin (ß3T) in Breast Cancer (BC) Using an Immunohistochemical (IHC) Assay in Studies of Ixabepilone (Ixa)
Authors: Roach C , Tanna V , Welcher R , Dumontet C , Sève P , Trifan O. 
Published: April 2011

Abstract
Background: Preclinical and retrospective clinical data suggest that ≥3T is a biomarker for poor prognosis and resistance to paclitaxel in breast cancer (BC). Preclinical studies have demonstrated that Ixa has activity in ≥3T over-expressing xenograft models resistant to taxanes and Ixa has shown clinical activity in BC patients with high ≥3T mRNA levels [Horak et al., J Clin Oncol 27:15s, 2009 (suppl; abstr 3587]. Dako and BMS have developed an IHC assay for detection of ≥3T to explore the potential of ≥3T as a biomarker to select BC patients that may benefit from Ixa. A neoadjuvant BC study comparing ixa to weekly paclitaxel employs this assay to assess ≥3T expression levels in patients’ tumor samples. Here we describe the application and initial validation testing of the assay in BC tumor samples.
Methods: An IHC assay employing a murine monoclonal anti-≥3T antibody (TUJ1) was developed to assess ≥3T expression in formalin-fixed paraffin-embedded tissue specimens. Banked specimens for initial testing included 233 cases of invasive BC.
Results: Patterns of ≥3T staining were typically homogeneous cytoplasmic staining with normal tissue elements such as endothelial cells and nerve cells serving as internal positive controls. Staining intensity was scored as 0, 1+, 2+ or 3+ with 0 being no visible staining and 2+ being equal to that of the internal positive control. Tumors with ≥50% cells staining ≥2+ were defined as positive for ≥3T based on previous published studies. Of the 233 breast tumors, 81 (35%) were positive and 152 (65%) were negative for ≥3T expression. HER2 status was defined for 128 of these specimens, with 37 of them (29%) found to be HER2 positive. Comparison of the samples with defined HER2 status demonstrated similar patterns of ≥3T expression, with 15 (40.5%) of the HER2+ samples being positive for ≥3T and 31 (34.1%) of HER2- specimens expressing elevated levels of ≥3T. In pathologist training exercises the inter-reader and intra-reader agreement of ≥3T scoring exceeded 85%. Antigen stability has been confirmed and other assay validation efforts are ongoing.
Conclusions: These findings are consistent with previously reported data on ≥3T expression in breast cancer, and support the use of this ≥3T IHC assay in studies with ixabepilone.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-08-06.
Abstract P4-08-11: Surface Plasmon Resonance (SPR) Analysis of COX-2 Expression for Early Detection and Progression of Breast Cancer and Evaluation of a Novel Class of Peptides as Its Inhibitor
Authors: Singh AK , Parshad R , Das SN , Dalal K , Dey S 
Published: April 2011

Abstract
Background: Cyclooxygenase-2 (COX-2) over expression has been observed in different cancers. Till date, COX-2 expression in breast cancer has been studied using RT-PCR and immunohistochemistry. The aim of this study was to use surface plasmon resonance (SPR) technology for determining the expression levels of COX-2 in breast cancer and design peptide inhibitors to evaluate their enzyme inhibition in vitro. Patients and methods: This case-controlled study was performed on 84 biopsy proven breast cancer patients having invasive ductal carcinoma and 40 age, sex and ethnicity matched healthy subjects. Blood and tissue samples were collected and serum and tissue lysate was prepared. Serum and tissue levels of COX-2 were evaluated using SPR where samples were allowed to flow over anti-COX-2 antibody immobilized sensor chip to obtain the respective RUs. Similarly, standard curve of COX-2 was prepared from cloned, expressed and purified protein to deduce the concentrations in different samples. To evaluate enzyme inhibition, fifteen peptides were synthesized using the solid and solution-phase peptide synthesis based on the active site structure of COX-2 and enzyme activity was evaluated spectrophotometrically.
Results: A significant increase in COX-2 levels in breast cancer patients 11.23-30.77μg/ml (Mean±SD=14.55±5.74) as compared to normal controls 2.13-5.16µg/ml (Mean±SD=3.71±0.86) (P&gt;0.0001) was observed. Serum COX-2 levels were significantly higher (P&lt;0.0061) in patients with lymph node involvement. More than 65% patients showed significantly (P&lt;0.0025) reduced COX-2 levels after chemotherapy. This was confirmed by western blot to check tissue level expression and spectrophotometrically to determine the activity of COX-2 at different stages. In addition, the peptides showed significant in vitro enzyme inhibition, and had a KD ranging from 1 X 10-8 to 1X10-10.
Conclusion: SPR is a useful proteomic technique for early detection and monitoring progession of breast cancer and peptides are a novel class of COX-2 inhibitors that can be further developed for anti-tumor therapy.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-08-11.
Abstract P4-09-02: Prognostic and Predictive Value of Centrally Reviewed Ki67 Labeling Index in Postmenopausal Women with Endocrine-Responsive Breast Cancer: Results from Austrian Breast and Colorectal Cancer Study Group Trial 8
Authors: Bago-Horvath Z , Rudas M , Jakesz R , Dubsky P , Singer CF , Dietze O , Greil R , Jelen A , Offner F , Lang A , Gruber C , Pöstlberger S , Gnant MFX , Filipits M. 
Published: April 2011

Abstract
Purpose: The aim of the present study was to assess the prognostic and predictive value of Ki67 labeling index in postmenopausal hormone receptor-positive early breast cancer patients who were treated with adjuvant tamoxifen or anastrozole after tamoxifen. Patients and Methods: We determined the expression of Ki67 by immunohistochemistry on whole tissue sections of breast carcinoma patients who had been enrolled in Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 8 and received tamoxifen for 5 years or tamoxifen for 2 years followed by anastrozole for 3 years. Ki67 labeling index was evaluated as continuous variable or dichotomized at 10%. Distant recurrence and death were analyzed using Cox models adjusted for clinical and pathological factors.
Results: High Ki67 labeling index was observed in 394 of 1587 (23%) tumors and was associated with poor outcome. Patients with high Ki67 labeling index had a significantly shorter distant recurrence-free survival (adjusted hazard ratio [HR] for distant recurrence 2.16, 95% confidence interval [CI] 1.43-3.25, P &lt; 0.001) and overall survival (adjusted HR for death 1.77, 95% CI 1.30-2.42, P &lt; 0.001) as compared to patients with low Ki67 labeling index. No interaction between Ki67 labeling index and treatment was observed (P = 0.84).
Conclusion: High Ki67 labeling index is an independent poor prognostic factor for distant recurrence and death in postmenopausal women with early-stage, hormone receptor-positive breast cancer but is not predictive for outcome of adjuvant treatment with either tamoxifen or tamoxifen followed by anastrozole.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-09-02.