bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P1-04-09: Extended characterization and target analysis of circulating epithelial cells in early and late breast cancer
Authors: Habets L , Bettina F , El-Ghali I , Körber W , Danaei M , Kusche M , Pachmann K 
Published: August 2014

Abstract
The MAINTRAC based (RBC lysis immunofluoretic detection and analysis on the Olympus ScanR system) finds more cells compared to enrichment based systems. The relative cheapness and low amounts of blood (1ml) allows frequent monitoring during early and late disease therapy steps. The clinical relevance of rising or falling counts of living (EPCAM+CD45negDAPIneg) CEC has been demonstrated in early and late disease by Pachmann. In addition we could show that these cells express stem cell, EMT and hypoxia markers. To our surprise we found high counts of this cell type in 90% of patients with fatty liver disease (NAFLD) in a non healthy non malignant control population. In a healthy control population in only 2% very few cells were found. Our conclusion so far CEC are a phenomenon driven by a hypoxic hyperacidic tissue situation shared by malignant and non malignant disease. The detectable cell population showed considerable morphologic heterogeneity urging us to introduce extended characterization and more comprehensive analysis. We therefore slightly adapted the basic MAINTRAC procedure to allow four color flowcytometric detection and analysis on the Amnis (EMD-Millipore) FlowSight device. The basic panel consists of (CD45PE, DAPI- blue, ALDH-FITC and EPCAMAF660). Extended Characterization is possible with different other panels including (HIF1, betacatenin, several pan- and single cytokeratins and CD44/CD24) and targets like ER, HER2, PDL-1 (immune checkpoint target) and phosphatidylserine. With this new approach we examined so far 66 patients with early and 53 patients with late disease. In early disease we found living CEC in 32 of 66 cases, with cell counts from 200 to 21642 per ml. In late disease in 27of 53 cases, cell counts varying from 481 to 97339 cells per ml. The percentage of living CEC varied from 0 to 60%. In all cases we found PDL-1 and ALDH1 expressed nearly exclusively on EPCAM+ cells demonstrating their suitability as a CEC specific marker. In cases with early relapse and rapid progression only few or none CEC are found especially in triple negative and HER2+ER negative cases. In these situations we find however EPCAM negative cell clusters. The IDEAS software allows comprehensive analysis of all visible phenomena including single cells, cell aggregates and exosomes. So far we still see no differences between “real” tumor derived CEC and “false positive” liver derived CEC. There is however preliminary evidence that CK20+ cells counts are much higher than EPCAM +counts in real compared with liver caused disease. A conclusive picture of these intruiging phenomena is not possible it seems however that the chosen way of extended characterization including new issues like cell clusters and exosomes should lead to a further elucidation of this biology and the use of CEC as a reliable “fluid biopsy”.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-04-09.
Abstract P1-04-12: Prognostic value of CTC detection by RT-PCR and the CellSearch system before first-line chemotherapy in metastatic breast cancer
Authors: Agelaki S , Mavroudis D , Politaki E , Apostolaki S , Kallergi G , Spiliotaki M , Georgoulias V 
Published: August 2014

Abstract
Background: CTCs have significant prognostic role in patients with breast cancer. Several techniques developed for CTC detection might help in monitoring treatment efficacy. The CellSearch System is the only clinically validated for use in patients with metastatic breast cancer (MBC). We have previously shown a concordance between RT-PCR for cytokeratin 19 (CK-19) and CellSearch System in the detection of CTCs in MBC. In this study we aimed to evaluate the prognostic value of CTC detection using these assays in the same cohort of patients. Methods: Blood was obtained from patients with MBC before the initiation of first-line chemotherapy. Different aliquots of the same blood sample were evaluated for the presence of CTCs by RT-PCR for CK-19 mRNA and by the CellSearch System. Disease progression and response to treatment were determined using standard clinical and imaging criteria. Results: In 142 patients with available clinical data, CTC evaluation was performed using both methods. Median age was 60.7 years (range, 23-82), 74.6% were post-menopausal and 16.9% were HER2-positive. CTCs were detected in 52.1% and 36.6% of patients by the CellSearch System (cut-offs ≥2 and ≥5 CTCs/7.5 ml, respectively) and in 38.7% by RT-PCR; 27.5% (cut-off ≥2) and 20.4% (cut-off ≥5) of patients were CTC-positive by both methods. Response rate was 40% in patients with CTCs detected by CellSearch (cut-off ≥2) vs 62% in CTC-negative patients; there was no difference in objective responses using the ≥5 cut-off, or using RT-PCR for CTC detection. Median PFS was 9.0 (cut-off ≥2) and 7.5 months (cut-off ≥5) for CTC-positive versus 20.0 (cut-off ≥2) and 18.7 months (cut-off ≥5) for CTC-negative patients using the CellSearch System (p = 0.0001). No significant difference in PFS was evident according to CTC detection by RT-PCR (10.0 vs 13.1 months, p = 0.253). Median survival was 24.7 vs 57.3 (cut-off ≥2) and 18.5 vs 53.7 (cut-off ≥5) months for CTC-positive vs CTC-negative patients using the CellSearch System (p = 0.0001); by RT-PCR, median survival was 30.3 and 50.1 months for CTC-positive and CTC-negative patients, respectively (p = 0.021). Conclusions: The detection of CTCs before first-line chemotherapy using either the CellSearch System or RT-PCR for CK-19 mRNA has significant prognostic value in patients with MBC.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-04-12.
Abstract P1-07-01: Histone deacetylase inhibitor entinostat reverses epithelial to mesenchymal transition of breast cancer cells by reversing the repression of E-cadherin
Authors: Sabnis GJ , Shah P , Kazi A , Gau Y , Chumsri S , Brodie A 
Published: August 2014

Abstract
Loss of ERa in breast cancer correlates with poor prognosis, increased recurrence rates and higher incidence of metastasis. In our previous studies, we have shown that histone deacetylase (HDAC) inhibitor entinostat (ENT) can upregulate ERα and aromatase in ER-negative cells and tumors, making them sensitive to aromatase inhibitors (AIs). In the current study, we are showing that ENT can also reverse epithelial to mesenchymal transition (EMT), which is considered to be a first step in the process of metastases formation. EMT is characterized by loss of intracellular adhesion (loss of E-cadherin); loss of epithelial markers such as cytokeratins and upregulation of mesenchymal markers such as vimentin; acquisition of fibroblast-like spindle morphology and increased motility. Various carcinomas undergo varying degrees of EMT and capacity to undergo EMT correlates inversely with levels of E-Cadherin. It is widely accepted that loss of E-cadherin is associated with more invasive phenotype. Epigenetic silencing of E-cadherin has been implicated in metastatic cell lines and invasive breast cancers.
Triple negative breast cancer cells such as MDA-MB-231 and Hs578T show a basal phenotype characterized by loss of E-cadherin expression and higher expression of mesenchymal markers such as N-cadherin, vimentin along with transcriptional repressors such as twist and snail.
In this study, we measured the effect of entinostat on the EMT. When MDA-MB-231 and Hs578T cells were treated with ENT, E-cadherin transcription was increased along with reduction in N-cadherin mRNA expression. Similar results were also seen in tumors of MDA-MB-231 and Hs578T xenografts treated with ENT (for 5 weeks and 2 weeks respectively). A dose dependent increase in E-cadherin was seen along with a dose dependent decrease in N-cadherin mRNA. Although, we did not observe any reduction in vimentin protein, phosphorylation of vimentin was increased and vimentin remodeling was changed as seen by immunofluorescence. We performed chromatin immunoprecipitation (ChIP) assay to measure the activation of E-cadherin promoter. Treatment of MDA-MB-231 and Hs578T cells increased the activation of E-cadherin promoter as seen by increased acetyl histones at the promoter region of E-cadherin. Twist and snail are known repressors of E-cadherin gene and we saw that ENT treatment reduced the association of twist and snail with the E-cadherin promoter. ENT was also able to downregulate twist, which may be responsible for reduced twist association with the E-cadherin promoter.
In summary, these findings suggest that HDAC inhibitor ENT can reverse EMT and may help reduce the formation of metastasis.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-07-01.
Abstract P1-07-04: A novel bioactive approach to inhibit leptin-induced epithelial-mesenchymal transition in breast cancer
Authors: Sharma D , Avtanski D , Nagalingam A , Kuppusamy P , Saxena N 
Published: August 2014

Abstract
Introduction: Molecular effects of obesity, a well-established risk factor for breast cancer progression, are mediated by adipocytokine leptin. Recent studies from our lab reveal that leptin induces epithelial-mesenchymal transition (EMT) and tumorsphere formation via concomitant activation of Akt/GSK3b and MTA1/Wnt1 signaling axes leading to β catenin activation. Given the important role of leptin in breast cancer growth and metastasis, novel strategies to antagonize biological effects of leptin are much desired. We showed previously that honokiol, a bioactive polyphenol from Magnolia grandiflora, inhibits breast carcinogenesis. The present study provides first evidence for the efficacy of honokiol against oncogenic effects of leptin including EMT.
Methods: Efficacy of honokiol to inhibit oncogenic effects of leptin was evaluated by using clonogenicity, anchorage–independent growth, matrigel-invasion and spheroid-migration assays. RT-PCR, western blot and immunofluorescence analyses were used to examine the molecular changes associated with EMT as well as underlying molecular pathways. Functional importance of MTA1/Wnt1/ β catenin axis was examined by using overexpression, phospho-deficient constructs and specific inhibitors. Finally, mouse xenografts, immuniohistochemical, RT-PCR and western blot analysis of tumors was used.
Results: Honokiol treatment circumvents leptin-induced EMT-associated phenotypic changes. A biochemical hallmark of EMT is loss of expression of epithelial markers with a concurrent increase in mesenchymal marker expression. Honokiol elicits increased expression of E-cadherin, occludin and keratin-18 (epithelial markers) and decreased expression of vimentin, fibronectin and N-cadherin (mesenchymal markers) in leptin-treated cells providing molecular evidence for reversal of EMT. Honokiol also inhibits expression and nuclear translocation of transcriptional effectors of EMT; Snail, Slug, Zeb1 and Zeb2. Analysis of underlying molecular mechanisms reveals that Honokiol effectively inhibits leptin-induced MTA1/Wnt1/ β catenin axis. Furthermore, using nontoxic doses of honokiol, we show that honokiol treatment effectively inhibits leptin-induced breast tumorigenesis in vivo. Analysis of breast tumors show that honokiol treatment reverses leptin-associated signaling (MTA1/Wnt1/ β catenin axis) along with decreased expression of mesenchymal markers and increased expression of epithelial markers.
Conclusions: In this study, we demonstrate for the first time that HNK is able to abolish leptin-induced EMT and provide in vitro and in vivo evidence for the integral role of a previously unrecognized crosstalk between honokiol and MTA1/Wnt1/ β catenin axis. Thus, HNK may be used as a non-toxic and non-endocrine rational therapeutic strategy for breast carcinoma in obese patients with high leptin levels.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-07-04.
Abstract P1-07-05: The cell fate factor DACH1 represses YB-1-mediated oncogenic transcription and translation
Authors: Pestell RG , Wu K , Chen K , Wang C , Jiao X , Wang J , Cai S , Addya S , Sorensen PH , Lisanti MP , Quong A , Ertel A 
Published: August 2014

Abstract
The epithelial-mesenchymal transition (EMT) enhances cellular invasiveness and confers tumor cells with cancer stem cell like characteristics, through transcriptional and translational mechanisms. The mechanisms maintaining transcriptional and translational repression of EMT and cellular invasion are poorly understood. The Drosophila homologue of DACH1, the Dac gene is a key member of the retinal determination gene network that specifies organismal development. The dachshund (dac), eya1, eyes-absent (eya), twin of eyeless (toy), teashirt (tsh) and sinoculues (so) are expressed in progenitor cells, contributing to development of the eye and genitalia. Loss of DACH1 expression contributes to the expansion of neural progenitors, muscle satellite cell differentiation and breast cancer stem cells. In recent studies Dachshund repressed breast cancer stem cell expansion. DACH1 expression is reduced in a variety of human cancers including prostate, ovarian and human breast cancer.
Herein, the cell fate-determination factor Dachshund (DACH1), suppressed EMT via repression of cytoplasmic translational induction of Snail by inactivating the Y box-binding protein (YB-1). In the nucleus, DACH1 antagonized YB-1-mediated oncogenic transcriptional modules governing cell invasion. DACH1 blocked YB-1-induced mammary tumor growth and EMT in mice. In basal-like breast cancer (BLBC) the reduced expression of DACH1 and increased YB-1, correlated with poor metastasis free survival. The loss of DACH1 suppression of both cytoplasmic translational and nuclear transcriptional events governing EMT and tumor invasion may contribute to poor prognosis in BLBC.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-07-05.
Abstract P1-07-06: E2F-1 directly induces HOXB9 accelerating breast cancer progression
Authors: Zhussupova A , Hayashida T , Takahashi M , Hiromitsu J , Kitagawa Y 
Published: August 2014

Abstract
Homeobox B9 (HOXB9) is a member of the class I HOX genes with highly conserved 61 amino acid homedomain motif, known to be overexpressed in breast cancer and related to tumor angiogenesis, EMT (Hayashida et al., PNAS 2010), and radio-resistance (Chiba et al., PNAS 2011), being significant prognostic factor in breast cancer patients (Seki et al., Ann Surg Oncol 2012). In this study, we investigated how HOXB9 expression is regulated in breast cancer cells.
First, we analyzed the upstream promoter region of HOXB9 gene to determine the important sequence for HOXB9 transcription. The multiple lengths of HOXB9 promoter region from -2898 to +1 were cloned into pGL3 luciferase reporter vector. Luciferase activities in MDA-MB231 cells which overexpress HOXB9 were analyzed and determined that the region from -404 to -392 played the critical role for the transcription of HOXB9 gene. Computer prediction system for the binding ability to critical sequence raised several candidate genes including E2F-1, PAX-5, P53, Sp-I, ETS-1 and NFY-A which are known to be related with cancer progression. It was confirmed that all these genes could induce HOXB9 overexpression, and especially E2F-1 was the strong inducer. Then, E2F-1-induced cell lines from MCF7, MCF10A and MDA-MB231were established and the correlation between E2F-1 and HOXB9 mRNA expression was determined. ChIP assay revealed that E2F-1 could directly bind to the critical sequence of HOXB9 promoter. Electrophoretic Mobility Shift Assay (EMSA) also indicated the binding ability of E2F-1. Finally, the tissue samples of 141 breast cancer patients, who undertook breast cancer surgery as a primary treatment, were stained with E2F-1 and HOXB9 antibodies, and correlation between HOXB9 and E2F-1 expression was confirmed with statistical significance (p&lt;0.001).
There are many publications describe the E2F-1 relation to the breast cancer patients prognosis. The context of this research displays the direct dependence of HOXB9 gene's expression through E2F-1, which might be suppressed by specific treatment, for example CDK4/6 inhibitor, and improve breast cancer patients’ survival.
To confirm these genes might demonstrate novel factors and/or signaling pathways playing role in breast cancer progression through HOXB9 axis.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-07-06.
Abstract P1-08-05: The prognostic significance of Ki-67 expression before and after neoadjuvant anthracycline-taxane-based chemotherapy in different biological breast cancer phenotypes
Authors: Zhang J , Zhang X , Yu F , Liu J , Zhang S , Hao X , Hou G 
Published: August 2014

Abstract
Purpose
This study was conducted to analyze the Ki-67 expression before and after neoadjuvant chemotherapy and clinical pathology characteristics of different biological breast cancer phenotypes at our center. A correlation study was performed between Ki-67 index change and the prognosis of different biological breast cancer phenotypes and prognosis in Northern China.
Methods
A regression analysis was performed on 213 patients with invasive breast carcinoma accepted NAC admitted to Tianjin Medical University Cancer Institute and Hospital of Breast Surgery from January 2007 to April 2008. These patients were subtyped by hormone receptor status and HER2 status.The Ki67 index (percentage of Ki67-positive cancer nuclei) were determined immunohistochemically. The prognostic value of the Ki-67 index for different biological breast cancer phenotypes disease-free survival (DFS) and overall survival (OS) was investigated by use of Kaplan–Meier curves and multivariable Cox regression.
Results
The overall pathologic CR (pCR) rate, defined as no invasive residuals in breast and axilla, was 17.8%. The highest pCR rate of 32.1% was observed in patients with HR-/HER2+ tumors, which is 11.4%, 20.0% and 26.2% in HR+/HER2- HR+/HER2-,HR+/HER2+and HR-/HER2- tumor respectively(P = 0.024).The Ki-67 expression of pre-NAC and post-NAC have prognostic significance in HER2- breast cancer, which have not significance different in HER2+ breast cancer:The best Ki-67 idex cut point of pre-NAC and post-NAC to predict long-term survival was respectively 20% and 10% in HR+/HER2- breast cancer. Moreover, patients with a higher Ki-67 index (&gt; cut-off point) showed significantly lower 5year-DFS/OS rates compared with those whose Ki-67 index were below cut-off point. In addition, Ki-67 index of post-NAC were independent prognostic factors for 5 year disease-free survival of patients with HR+/HER2- breast cancer. Ki-67 change between pre- and post-NAC as an independent prognostic factor independently predict prognosis in the patients who have not achieved pCR, while the best cut-off point was 18%.
Conclusion
The Ki-67 index of pre- and post-NAC could predict the prognosis in the patients with HER2- breast cancer. Moreover, the Ki-67 change between pre- and post-NAC was an independent prognostic factor in the patients who have not achieved pCR.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-05.
Abstract P1-08-13: Predictive value of intrinsic luminal subtypes in premenopausal women with endocrine-responsive early breast cancer: Results from Austrian breast and colorectal cancer study group trial 5
Authors: Bago-Horvath Z , Rudas M , Jakesz R , Greil R , Dietze O , Suppan C , Lax S , Singer CF , Filipits M , Gnant M 
Published: August 2014

Abstract
Purpose: The aim of the present study was to assess the predictive value of intrinsic luminal subtypes in premenopausal hormone receptor-positive early breast cancer patients who received adjuvant endocrine treatment or chemotherapy.
Patients and Methods: Intrinsic luminal breast cancer subtypes were centrally assessed by immunohistochemistry on whole tissue sections of breast cancer patients who had been enrolled in the Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 5 and received either 3 years of goserelin plus 5 years of tamoxifen or six cycles of cyclophosphamide, methotrexate, and fluorouracil (CMF). Luminal A disease was defined as Ki67 &lt;20%, Luminal B/HER2-negative subtype as Ki67≥20%. Luminal B/HER2-positive subtype displayed HER2 overexpression by IHC or amplification by ISH. Recurrence-free survival (RFS) and overall survival (OS) were analyzed using Cox models adjusted for clinical and pathological factors.
Results: 155 (36%), 217 (50%) and 59 (14%) of 431 tumors were classified as Luminal A, Luminal B/HER2-negative and Luminal B/HER2-positive, respectively. Luminal B subtypes were associated with poor outcome. Patients with Luminal B tumors had a significantly shorter RFS (adjusted hazard ratio [HR] for recurrence: 2.29, 95% confidence interval [CI] 1.44-3.64, P &lt; 0.001) and OS (adjusted HR for death: 3.97, 95% CI 1.94-8.08, P &lt; 0.001) as compared to patients with Luminal A disease. No interaction between intrinsic luminal subtypes and treatment was observed (test for interaction: P = 0.42 for RFS; P = 0.32 for OS). Combination endocrine treatment with goserelin/tamoxifen tended to be more effective than CMF chemotherapy in both luminal A and luminal B subtypes.
Conclusion: Intrinsic luminal subtype is an independent prognostic factor for recurrence and death in premenopausal women with early-stage, hormone receptor positive breast cancer but is not predictive for outcome of adjuvant treatment with either goserelin/tamoxifen or chemotherapy with CMF.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-13.