bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P2-19-05: Inhibitor Screening Utilizing Human Kinase Multiplex Arrays
Authors: Sahli N , Finkel D , Lindberg M , Buehl C , Stoesz S , Campos-Gonzalez R , Brumbaugh K. 
Published: April 2011

Abstract
Protein kinase pathways show dysregulation in cancer, usually displaying increased activation. Kinases are typically influenced by extracellular signals, and many kinases are now considered to be therapeutic targets. Upon phosphorylation, kinases regulate signaling pathways that effect cellular processes. Individual cell lines, including cancer cells, express different kinase activation patterns. Previously, we have shown the phospho-tyrosine kinase profile of MDA-MB-453 cells, most notably the Erb family of Receptor Tyrosine Kinases, to be hyperactivated in this breast cancer cell line. Using a new kinase multiplex plate-based array, we have seen similar activation of multiple intracellular kinases, such as Akt and Erk. As a result, the MDA-MB-453 cells were selected for treatment with a kinase inhibitor library (Enzo, Catalog # BML-2832-0100). While several of the inhibitors block kinase activation, other inhibitors block proteins that are upstream from their kinase target. After incubation with the inhibitor library, a multiplex kinase array was utilized to screen the effects of the inhibitors on the MDA-MB-453 cells. In contrast with MCF-7 cells, MDA-MB-453 breast cancer cells showed a basal hyperactivation of the Akt and GSK-3β kinases. The activation of Akt could be selectively inhibited by SB-203580, SB-202190, ML-7, and BML-265 inhibitors. The results obtained in the multiplex assay were verified by singleplex ELISA for Akt. Hence, the data collected with the multiplex assay provide a rapid analysis of the inhibitors’ effects on defined signaling pathways. This allows for a faster identification of kinase inhibitors that may affect cancer cells.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-19-05.
Abstract P3-02-06: Correlation of BCL-2 and Apoptosis on Circulating Tumor Cells and Breast Cancer Tissue
Authors: Maranci V , Thomas DG , Brown M , Budd TG , Doyle G , Hayes DF , Griffith KA , Smeage J. 
Published: April 2011

Abstract
Background: Circulating Tumor Cell (CTC) levels are prognostic markers in metastatic breast cancer (MBC). Further phenotypic characterization of CTC may provide an opportunity for non-invasive evaluation of predictive and prognostic markers. Apoptosis is a common form of chemotherapy-induced cell death. Monoclonal antibody (MAb) M30 recognizes a neo-epitope on fragmented cytokeratin and is a marker of apoptosis. BCL-2 is an anti-apoptotic marker and may predict for resistance to anti-neoplastic therapy. We have previously reported the results of a pilot clinical trial to estimate M30 and BCL-2 expression in CTC from patients with MBC. The current study was performed to correlate the expression of both BCL-2 and M30 in CTC with breast cancer tissue samples. Methods: Of the 85 patients in the original pilot study, 52 (61%) had evaluable tissue available (39 primary and 12 metastatic lesions; one unknown) and were included in this analysis. CTC were collected at baseline and were isolated, enumerated, and phenotypically characterized for M30 and BCL-2 using the CellSearch® System. CTC phenotype is reported as percentage of cells with positive staining. Tissue Microarrays (TMA) were constructed and immunohistochemically stained for M30 (anti-Cytodeath™ M30, Roche) and Bcl-2 (M-0887, DAKO) and scored using the Allred method.
Results: Tissue staining for M30 and Bcl-2 were inversely correlated. Of the 52 patients with TMA available for investigation, 22 (42%) had &gt; 5 CTC/7.5 ml whole blood. A positive, but non-significant, correlation was observed between increasing numbers of CTC levels and tissue BCL-2 Allred Score (Spearman r=0.36; p=0.1310). M30 staining was detected in ≥10% of CTC in 20 of the 22 (91%) patients with elevated CTC, while BCL-2 staining was detected in ≥10% of CTC in 19 of the 22 (86%) patients. Little if any correlation was observed between CTC M30 and tissue M30 expression, however CTC BCL-2 was positively correlated with tissue Bcl-2 expression (Spearman r=0.47; p=0.0440). Conclusions: BCL-2 and M30 can be characterized in both CTC and breast cancer tissue. The likelihood of having elevated CTC may be associated with higher BCL-2 expression, and CTC BCL-2 was associated with tissue BCL-2 expression. Although the clinical implications of these findings are unknown, they offer the opportunity for future trials to investigate whether CTC BCL-2 and M30 might be useful to identify patients with cancers that are resistant to standard therapies. Furthermore, CTC BCL-2 and M30 might be monitored in clinical trials of novel, apoptosis-inducing therapeutic agents.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-02-06.
Abstract P3-09-04: Exosome-Specific microRNA Signatures in Combination with Characteristic Surface Markers on the Circulating Exosomes Themselves Provide New Insights into the EMT
Authors: Paulaitis M , Guzman N , Agarwal K , Saji M. 
Published: April 2011

Abstract
Background: Recent discoveries have established that cancer tumors exhibit distinct microRNA (miR) expression profiles compared to normal tissues. Detection of miRs in the peripheral blood of cancer patients is possible due to their high stability. This stability is attributed to encapsulation of the miRs inside microvesicles (MVs) where they are protected from endogenous RNase activity in circulation. The tumor cell-secreted MVs of primary interest are circulating exosomes, a subpopulation of MVs distinguished by their relatively small size: 40 to 100 nm in diameter. The isolation of circulating tumor-derived exosomes from the other cell-secreted MVs in peripheral blood is the critical barrier to successful development of a robust assay for cancer-specific miR signatures. We describe the development of such an assay to: (1) capture/isolate circulating exosomes based on characteristic surface markers that correlate with different cell types, and (2) characterize exosome-specific miR signatures based on these surface markers.
Materials and Methods: We apply sequential ultracentrifugation to isolate cell-secreted MVs, followed by asymmetric flow field-flow fractionation to selectively separate exosomes based on their intrinsically small size and characteristic surface markers. Isolated exosomes are then screened for surface markers by selectively capturing them on microarrays printed with antibodies against a library of known cell-surface markers for breast cancer. For the assessment of exosome-specific miR signatures, we have devised an antibody microarray assay that also enables the in situ characterization of miR profiles of the captured exosomes locally by qRT-PCR analysis confined to subarrays of printed spots in 50-µl microwells of a custom-designed, multi-well microarray. Assay development has been carried out using two human cancer cell lines (MCF 7 and MDA MB 231) and a non-malignant cell line (MCF 10a) representing pre-malignant cells and cancer of the breast at different epithelial and mesenchymal states. Results: We have used light scattering and cryo-transmission electron microscopy to characterize the size, size distribution, and morphology of exosomes as a sub-population of the secreted MVs from each of the three cell lines. We have also screened for surface markers by selective capturing the exosome sub-populations derived from these cell lines on antibody microarrays, and correlated these surface markers with the miR content of MVs using qRT-PCR analysis.
Discussion: Our results demonstrate that correlating the miR signatures of circulating exosomes with characteristic surface markers on these exosomes leads to robust distinctions between the three cell lines that could not be achieved using either analysis or characterization alone.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-09-04.
Abstract P3-09-05: Let-7 microRNAs Regulate a Novel Variant of Estrogen Receptor Alpha, ER-≥36, Nongenomic Estrogen Signal Pathway, and Tamoxifen Resistance in Breast Cancer
Authors: Zhao Y , Deng C , Lu W , Xiao J , Recker RR , Guishan GG. 
Published: April 2011

Abstract
Background: MicroRNAs (miRNAs) have important regulatory functions in breast cancer tumorigenesis. We previously found that let-7 miRNAs were significantly downregulated in formalin-fixed paraffin-embedded (FFPE) breast cancer tissues.
Material and Methods: QRT-PCR was used to check let-7 family miRNAs in FFPE tissues and breast cancer cell lines as well as expression of estrogen receptor (ER)-≥36, a variant of ER-≥66, after let-7 miRNA transfecton. Immunoblot analysis was employed to check protein expression in FFPE tissue and breast cancer cell lines. Luciferase reporter assay was used to detect direct regulation of let-7 miRNA on ER-α expression. MTT assay was applied for cell proliferation after transfection of let-7 miRNAs. Results: We found that there was an inverse correlation between the expression of ER-≥36 and several members of let-7 family miRNAs in the FFPE tissue set. Let-7 miRNA sequences match sequence in the 3’ untranslated region (3’ UTR) of ER-≥36, indicating ER-≥36 may be a target of let-7. Cotransfection of let-7 mimics with ER-≥36 3’ UTR luciferase construct decreased the activity of reporter gene. Conversely, let-7 inhibitors enhanced the reporter gene activity. Transfection of let-7 mimics inhibited both the mRNA and protein levels of ER-≥36 and further inhibited the non-genomic estrogen pathway mediated by ER-≥36 in MDA-MB-231 cells. On the contrary, transfection of let-7 inhibitors enhanced the ER-≥36 expression at both mRNA and protein levels in 184A1 cells. The high expression of ER-≥36 in tamoxifen resistant MCF7 cells can be inhibited by transfection of let-7 mimics and sensitivity toward tamoxifen is enhanced.
Conclusion: Let-7 miRNAs regulate the expression of ER-≥36, which is involved in non-genomic estrogen pathway and tamoxifen resistance. Let-7 could be therapeutic target for breast cancer treatment.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-09-05.
Abstract P3-10-03: The Implication of Androgen Receptor Expression in Breast Cancers
Authors: Park S , Koo JS , Kim MS , Park HS , Lee JS , Lee JS , Kim SI , Park B-W. 
Published: April 2011

Abstract
Purpose: To evaluate the implication of androgen receptor (AR) expression in patients with breast cancer.
Methods: Immunohistochemically AR expression was investigated in 931 invasive breast cancers prepared in tissue microarray blocks that were collected in between November 1999 and August 2005. The clinicopathological features, disease-free (DFS) and overall survival (OS) were analyzed using chi-square test, Kaplan-Meier methods, and Cox's models. Tumors with ≥10% nuclear-stained cells were considered to be positive for AR, estrogen receptor (ER), and progesterone receptor (PR) expression and 3+ with immunohistochemical staing or FISH + were considered positive for HER2 expression.
Results: The positive expression of AR, ER, PR, and HER2 was 58.1%, 72.2%, 61.4%, and 24.7% of all patients, respectively. AR expression was significantly associated with an older age at diagnosis (p=0.007), smaller tumor size (P&lt;0.001), well differentiation (P&lt;0.001), higher hormone receptors expression (P&lt;0.001), non-triple-negative breast cancers (non-TNBC) (P&lt;0.001), and lower proliferation index (P&lt;0.001). In ER-negative cancers, AR expression was associated with well differentiation, lower Ki-67 index, and distinctively HER2 overexpression. With median follow-up duration of 72.7 months, AR was significantly associated with better DFS (p=0.023) and OS (p=0.048) and the association was same in ER-positive cancers but was disappeared in ER-negative cancers. In Cox's models, AR expression was an independent prognostic factor for DFS (hazard ratio, 0.654; p=0.049), but it did not reach a statistical significance for OS (hazard ratio, 0.647; p=0.119) even in ER-positive tumors. Interestingly, tumors with molecular apocrine features (ER-negative, AR-positive, and HER2-positive) showed a trend of poorer outcome with AR expression, but there was no impact of AR expression on the survival in 156 patients with TNBC.
Conclusions: AR expression is significantly associated with favorable clinicopathological features and better outcomes in ER-positive cancers but tumors with molecular apocrine features showed a trend of poorer outcome with AR expression. These results suggest that AR expression could be an additional marker for endocrine responsiveness in ER-positive tumors and a target molecule for the molecular apocrine tumors.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-03.
Abstract P3-10-04: An Integration of Biological and Pathological Marker Panel in the TEAM Pathology Sub-Study: The Impact of Different Parameters on Risk Estimation of Relapse at Both 2.75 and 5 Years
Authors: Bartlett JMS , Stocken D , van de Velde CJH , Brookes CL , Robson T , Hasenburg A , Hille ETM , Kiebeck D , Markopoulos C , Mallon EA , Dirix L , Campbell FM , Seynaeve C , Rea DW. 
Published: April 2011

Abstract
Background: Recent evidence confirms the importance of both biological and pathological risk markers in predicting early relapse for breast cancer patients treated with endocrine therapy. Most studies use a two step process integrating biological markers into a “biological predictor (e.g. Oncotype Dx, “IHC4” etc) followed by assessment of the predictive value of such tests in the context of pathological markers (grade, nodal status etc). We have taken a one step process integrating both biological and pathological markers into a single model to assess key factors for predicting outcome at 2.75 years and 5 years of endocrine therapy; to inform choices between switching, upfront and extended adjuvant treatment with AIs. Patients & Methods: Pathology blocks from 4598 TEAM patients were collected and tissue microarrays constructed. Quantitative analysis ER, PgR, Ki67, HER1, HER2, and HER3 was performed centrally. A prognostic model, integrating data from biological and pathological markers was created to assess risk (disease-free survival) after 2.75 and 5 years of follow up in the TEAM trial.
Results: Of 4595 eligible cases samples received, 16 were excluded, and 3993 had complete biomarker data for all markers for the final biomarker analysis. In univariate analysis nodal status, grade, size, age at diagnosis, HER1, HER2, PgR, ER and Ki67 were all prognostic. At 2.75 years nodal status, age, PgR histoscore, size, grade, HER2, ER histoscore and HER1 positivity were significant prognostic variables (ranked by WaldX2 statistic), Ki67 and HER3 were not included in this model. At 5 years median follow up; age, nodal status, size, PgR histoscore, grade, Ki67, HER2, and HER1 positivity were significant prognostic variables (ranked by WaldX2 statistic), ER and HER3 were not included in this model. Conclusion: Combined biological and pathological marker panels are of significant value in predicting early relapse in breast cancer patients treated with endocrine therapy, however duration of follow-up may impact on the inclusion of variables in the model. This provides significant information relevant to the choice of different adjuvant endocrine therapies.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-04.
Abstract P3-10-05: Triple Negative Breast Cancer: Clinical Characteristics in the Different Histological Subtypes
Authors: Dreyer G , Vandorpe T , Forceville K , Neven P , Smeets A , Janssen H , Brouwers B , Deraedt K , Paridaens R , Wildiers H. 
Published: April 2011

Abstract
Purpose: To study clinical characteristics of different histological subtypes of triple negative breast cancer (TNC). TNC are often considered as one group, but consist of many different histological subtypes with different biology and behavior. We decided to analyze a large cohort of TNC in a single centre and focus on clinical behavior including age distribution, occurrence of lymph node invasion, and outcome in different histological subtypes.
Patients and methods: We used data from the breast cancer database from UH Leuven, containing 479 newly diagnosed TNC from January 1999 till November 2009, of which 396 received upfront surgery, 64 neoadjuvant chemotherapy, and 18 with metastases at diagnosis. Univariate and multivariate relations were studied between age, lymph node status, histological subtype, treatment and outcome.
Results: Apocrine and invasive lobular TNC occur more often in older patients compared to IDC-NOS (resp. P = 0.0004, P = 0.0496), the latter being by far the most common subtype (80,2%).
Although TNC are considered as an aggressive subtype of breast cancer with a bad prognosis, only 35.1% have lymph nodal involvement. There were no significant differences in nodal invasion between subtypes, but most subtypes contained few patients. In contrast to previous reports, 7/14 (50%) of apocrine TNC had lymph node involvement.
DFS was different in different histological subtypes, but most of the histological subgroups contained insufficient numbers to be able to draw firm conclusions.
Within the histologically ‘homogeneous’ IDC-NOS group with primary surgery and outcome data (n=308), DFS with 3.5 year median FUP decreased with increasing age, but chemotherapy and also radiotherapy were much less frequently given with increasing age (in age &lt;50, 50-69, 70+ no chemo in 2/135, 25/126, 39/44; no radiotherapy in 14/135, 22/126, 12/44 respectively). In multivariate analysis, increasing age (HR for 10 yr increase: 0.68 (0.52-0.89)), presence of lymph node involvement (HR 2.17 (1.24-3.77)), administration of chemotherapy (HR 0.28 (0.12 — 0.67)), and radiotherapy (HR 0.39 (0.20-0.73)) were significant predictors of relapse.
Conclusion: TNC is not a uniform disease. Different histological subtypes have different age distribution and outcome. The prognosis of the most common group of TNC, IDA-NOS is unexpectedly better in older patients, but this is counterbalanced by the significantly decreased use of chemotherapy and radiotherapy in this population.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-05.
Abstract P3-10-10: Progesterone Receptor-Negative Status Predicts Poorer Outcomes in Estrogen Receptor-Positive Patients Treated with Selective Estrogen Receptor Modulators
Authors: Chagpar AB , McMasters KM , Edwards MJ 
Published: April 2011

Abstract
Introduction: While estrogen receptor (ER) status has been used as the primary factor influencing the use of selective estrogen receptor modulators (SERMs), a number of ER-positive patients will relapse on this therapy. It has been postulated that SERM-resistance may be modulated by aberrant growth factor signaling, for which progesterone receptor (PR) status may be a surrogate. PR status, however, is frequently not considered as a strong prognostic or predictive marker in ER-positive patients. We sought to define the effect of PR-status on outcomes in such patients treated with SERMs.
Methods: The North American Fareston versus Tamoxifen Adjuvant (NAFTA) Trial is an investigator-initiated, multicenter, prospective trial that randomized patients with resected hormone-receptor positive breast cancers to receive either adjuvant tamoxifen (20 mg po daily) or toremifene (60 mg po daily) for five years. Between July 1998 and December 2002, 1692 ER-positive positive patients were randomized. PR was missing for 15 patients. The remaining 1677 (1424 PR-positive and 253 PR-negative) patients formed the cohort of interest for this analysis. The impact of PR status on overall (OS) and disease-free survival (DFS) was assessed using Kaplan Meier and Cox Proportional Hazards modeling. Results: The median patient age was 67 (range; 41-100) with a median tumor size of 1.2 cm (range; 0.01-14.00 cm). PR-positive status was correlated with node-negative disease (91.3% vs. 87.0%, p=0.035) and low grade tumors (34.9% vs. 27.7%, p=0.009), but was not associated with tumor size (p=0.533) or patient age (p=0.067). With a median follow-up of 59 months, the 5-year actuarial OS and DFS of PR-negative patients was significantly worse than that of PR-positive patients (88.5% vs. 94.3%, p=0.009 and 86.1% vs. 92.5%, p=0.003, respectively). Controlling for tumor size, grade and lymph node status, PR-negative status continued to predict worse DFS (OR=1.561; 95% CI: 1.013-2.406, p=0.043), although it lost significance in terms of OS (OR=1.619; 95% CI: 0.980-2.674, p=0.060).
Conclusion: PR-negative status is an independent predictor of worse DFS in ER-positive patients treated with SERMs. These data support the hypothesis that PR-negative status may signal a higher rate of SERM-resistance, and should be considered in adjuvant therapy decisionmaking.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-10.
Abstract P3-10-14: The Prognostic Significance of mib-1 (according to St Gallen Criteria) in Patients (pts) with Early Invasive Breast Cancer (EIBC) and Its Relationship with other Biologic Parameters in a Monoinstitutional Series
Authors: Ferro A , Caldara A , Eccher C , Triolo R , Aldovini D , Cuorvo LV , Frisinghelli M , Fasanella L , Berloffa F , Barbareschi M , Galligioni E 
Published: April 2011

Abstract
PURPOSE: To examine interactions between mib-1 and clinical-pathological markers and their impact on outcomes. PATIENTS AND METHODS: Mib-1 was identified by immunohistochemical staining in 3402 EIBC pts treated from 1995 to 2008. Mib-1 was defined as low (15%), intermediate (16%-30%) and high (&gt;30%) value. Correlation between mib-1 and age, tumor size (T), nodal status (N), ER, PR, HER-2, grading (G) was performed by X2 test. Log-rank test and Cox regression model were performed to test mib-1 as prognostic factor for overall survival (OS) and Disease Free Survival (DFS) and its correlation with other known prognostic factors.
RESULTS: Median mib-1 was 25%. Mib-1 was low in 1135, intermediate in 1094 and high in 1173 pts. Median age was 62 years. High mib-1 was significantly (P&lt;0.001) associated with younger age, ductal type, greater size, positive N, poor G, absent or low ER and/or PR expression level, positive HER-2, triple negative subtypes, larger use of chemo±hormonotherapy. High mib-1 was a significant predictor of worse
DFS and OS (P&lt;0.001). At median follow up of 42 months DFS and OS were 83.1 and 84.8% respectively in high, 88.9 and 89.3% in intermediate, 92.7 and 93.6% respectively in low Mib-1 group. There were 401 relapses (12%): 197 (6%) in high, 121 (3.5%) in intermediate and 83 (2,5%) in low mib-1 group. BC deaths were 368 (10.8 %): 178 (5.2%) in high, 117 (3.4%) intermediate and 73 (2.2%) in low mib-1 group. High mib1 was predictive of poorer prognosis in &gt; 40 years pts (P&lt;0.001), small tumors (T1) (P&lt;0.001), N+ (P&lt;0.001) and N0 (p=0.007), G1 (p=0.02), ER+ and/or PR+ (P&lt;0.001), negative HER-2 (p=0.007), ductal type (P&lt;0.001) respect to intermediate and low Mib1. At Cox regression analysis Mib1 showed independent prognostic value. According to the median value, high mib-1 (&gt;25%) resulted a prognostic factor of worse outcome (P&lt;0.001, HR=1.81, 95% CI=1.42-2.29), along with positive lymph nodes (P&lt;0.001, HR=2.14, 95% CI=1.67-2.74), age&lt;40 yrs (p=0.002, HR=1.93, 95% CI=1.30-2.86), greater tumors (P&lt;0.001, HR=3.11, 95% CI=2.21-4.39), and ER/PR positive tumors (P&lt;0.001, HR=1.742, 95% CI=1.305-2.327).
CONCLUSIONS: In our experience, Mib-1 confirms to be a significant prognostic biomarker for DFS and OS in EIBC, providing additional information besides other clinico-pathological parameters to help decision-making of treatment.
Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-14.