bluuebunny/crossref_metadata_2025_split huggingface dataset:
Monitoring cytokeratin19 (KRT19) levels as response of circulating tumor cells (CTC) to neoadjuvant chemotherapy (NAC) in breast cancer (BC).
Authors: Guerrero-Zotano AL , Garcia-Casado M , Fernandez-Serra A , Lopez-Guerrero J , Gavila J , Climent M , Ruiz A , Guillem V 
Published: December 2010

Abstract
Abstract #5022
Background: KRT19 mRNA+ cells before the initiation of adjuvant chemotherapy has been shown as an independent prognostic factor for disease recurrence in patients with early BC. Its usefulness for monitoring response to NAC it is not clear. We have studied changes in KRT19 levels during NAC and correlated it with response. Material and Methods: We analyzed a prospective series of 70 BC patients treated with NAQ. Stage II (45%), Stage III (55%), HR negative (43%), HER-2 positive (42%). Treatment was based on sequential anthracyclines and taxanes. Trastuzumab was given concomitant in HER2 disease. KRT19 level expression was measured from lymphocytes of peripheral blood using a quantitative RT-PCR. The analysis was performed at diagnosis (baseline), before taxane administration, at the end of NAC, and after surgery. On the other hand, 27 healthy donors and 18 metastasic BC patients were also analyzed in order to determine a cut-off value for CTC positivity according to receiver operating characteristics [ROC] analysis. Association between variables was performed by Pearson's Chi-squared and Fisher's tests. Results: We observed a differential rate in KRT19 expression, between the control and the metastasic series being higher in the last group (U-Mann-Withney test, p=0.010). According to the selected cut-off value (AUC 0.272; p=0.010), we detected CTC in 46/70 (65.7%) patients at baseline, 22/40 (55%) prior to taxane administration, 36/50 (72%) at the end of NAC and 13/17 (76.5%) after surgery.Forty out of 57 patients (70.2%) responded to NAC(15 CR and 25 PR). Decreased in CTCs levels during treatment wasn't associated with response, on the contrary, residual CTCs were detected after surgery in all those cases that responded (p=0.005). The presence of CTCs after completion of chemotherapy, previous to surgery, was inversely related to tumor stage (p=0.012). Negative tumors for HER2 and tumors with high levels of ER expression maintain the CTC levels detected at diagnosis (p=0.044 and p=0.057 respectively). DISCUSSION: We weren't able to find a correlation between response and decrease in CTCs levels. The presence of residual CTCs in responding cases could suggest a release of tumor cells into blood by NAQ and surgery The prognostic factor of residual CTCs needs to be addressed with longer follow up, and taking into account the effect of others therapies. This study is financed by the grant GVA07/130.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5022.
Practical use of circulating tumor cells for the management of subsets of metastatic breast cancer patients in a community oncology practice.
Authors: Dropkin EK , Graham ML 
Published: December 2010

Abstract
Abstract #5024
Background : An elevated Circulating Tumor Cell Count (CTC) &gt;5 in 7.5 ml of blood predicts worse outcome in metastatic breast cancer (MBC) patients (pt). While the FDA has approved use of this test, its systematic use in practices has been uncommon. We have measured CTC's in all MBC pts in our community oncology practice receiving hormonal therapy or chemoRx +/- biologic therapies to determine if a strategy to treat all pts to a sustained CTC count of &lt; 5 could decrease the death rate, and if recognized molecular subsets of breast cancer [ER+, HER2 (H2)-; Triple Negative; H2+] have different patterns of CTC behavior. Methods: Initially serial CTC levels were measured by the CellSearch technology (Veridex, LLC; Warren NJ) using two commerical laboratories in all pts whether beginning a new therapy with initial recurrence or progression, or if already underway with treatment. Pts had CTC's measured from Nov 20, 2006 thr. June 10, 2008, an 18 mo. period Over time, the majority of pts have begun new treatments with progression, or with new diagnoses. If pts with elevated CTC's improved to &lt;5 for a sustained 3-4 month period of time, we sought less morbid hormonal therapies if ER+, or lower toxicity ChemoRx +/- biologic therapy if ER- or hormone refractory. Results: 376 tests for CTC's were carried out in 72 pts for an average of 5.2 tests per pt (range 1-19). The distribution of the 72 pts according to the three molecular subtypes, the frequency of those measuring &gt;5, &gt;10 or &gt;100 CTC's at any time, and the associated death rates for each group, are shown in Table 1.&#x2028; &#x2028; High numbers of CTC's were seen in each of the three subsets of MBC, and each subtype also had pts with no elevated levels. 2/3 of deaths occurred in pts with 100 or more CTC's at some point in their treatment course. Of pts with these high levels, more than 50% eventually died, while only 3% died over 18 months if serial measurements showed no levels above 5 CTC's. Deaths from MBC in the practice have been reduced from 20 in 2005 and 15 in 2006 to 9 in the 18 mo. during which the CTC's have been uniformly applied in 2007 and 2008. Conclusion: Very high levels of CTC's &gt; 100 constitute an oncologic emergency. The overall death rate in a community oncology practice may be reduced by uniform application of CTC technology.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5024.
Isolation of circulating tumour cells in patients with metastatic breast cancer by filter technology.
Authors: Elst H , Van Der Auwera I , Vermeulen P , Prové A , Van Dam P , Huget P , Dirix L 
Published: December 2010

Abstract
Abstract #5026
Introduction. Metastasis is the leading cause of cancer-related death. For several decades, numerous attempts have been made to develop reliable assays enabling the detection and enumeration of circulating tumor cells (CTC) in the peripheral blood. The LeukoLOCK™ Total RNA Isolation System (Ambion®) combines leukocyte depletion filter technology intended to isolate leukocytes from whole blood, with RNALater® as a means of stabilizing the RNA in the cells captured on the filter. We hypothesized that this innovative system would also allow for the isolation of carcinoma cells by filtration, because of their larger size in comparison to peripheral blood leukocytes. The RNA extracted from the filter device could thus be used for further quantification of CTCs by qRT-PCR.&#x2028; Materials and methods. The CellSearch™ Circulating Tumor Cell Test (Veridex®) was used to enrich and enumerate CTCs in all of the blood samples. The CellSearch System™utilizes iron nano-particles (ferrofluid) conjugated to EpCAM to capture CTC. Immuno-fluorescent staining for cytokeratins (CK) 8,18,19 and the nucleus (DAPI) is used to positively identify intact CTC. CTC are defined as nuclear cells, expressing cytokeratins and lacking CD45 expression. Blood samples of 5 patients with known elevated CTC were screened again after filtration with the LeukoLOCK™filter device to ensure that CTC did not slip through the filter (Table 1). The cell count obtained with the Veridex system was compared in these 5 samples, with the quantification CK19 and mammaglobin (MAM) transcripts obtained from the cells retained by the filter. In a further study on 79 patients with metastatic breast cancer, CTCs were enumerated using LeukoLOCK™Total RNA Isolation System (Ambion®) as a means of RNA isolation and compared with the Veridex system. qRT-PCR was performed to detect the presence of CTC transcripts for CK19 in the total RNA obtained from the cells captured on the filter in all of the 79 cases.&#x2028; Results. In only one sample (PAT113) could one CTC be detected in the filtered blood. The values obtained for RGE for both CK19 and MAM are in agreement with that observation.&#x2028; &#x2028; A significant correlation was found between CTC levels and qRT-PCR normalized relative gene expression (nRGE) values for CK19 originating from the total cell population captured by the LeukoLOCK™filter device (n=79, r=0.446, p&lt;0.001).&#x2028; Conclusion. The LeukoLOCK™ system does not only enrich for leukocytes but can be considered as a reliable system for the enrichment and stabilization of CTC in patients with MBC.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5026.
Cyclin overexpression inhibits Smad 3 tumor suppression in breast cancer cells.
Authors: Zelivianski S , Cooley A , Jeruss JS 
Published: December 2010

Abstract
Abstract #5033
Introduction: The transcription factor Smad 3, a member of the TGFβ signaling cascade, contributes to G1 cell cycle arrest in breast cancer cell lines. Overexpression of the cell cycle regulatory protein cyclin E, and its low molecular weight (LMW) counterparts, has been associated with poor prognosis in breast cancer. Cyclin E has been shown to mediate phosphorylation within or near the linker region of Smad 3 by cyclin dependent kinase 2 (CDK 2), and to inhibit Smad 3 activity in normal mouse fibroblasts and epithelial cells. We hypothesize that overexpression of cyclin E may exert tumorigenic effects in breast cancer cell lines through the functional inhibition of Smad 3 mediated by CDK 2 phosphorylation.&#x2028; Methods: Endogenous levels of Smad 3 and phosphorylated Smad 3 in parental, vector only control, full length (FL), and two LMW forms of cyclin E overexpressing MCF-7 breast cancer cell lines were determined by western blotting. To elucidate the impact of cyclin E overexpression on Smad 3 function, constructs containing an empty vector, WT Smad 3, Smad 3 with a CDK phosphorylation site mutation within the linker region (Thr178), and outside the linker region (Thr8) were co-transfected with a Smad 3 reporter into the panel of MCF-7 cells. CDK2 or scrambled control siRNA was also transfected into the study panel of cells. Smad 3 function was then evaluated by luciferase reporter assays.&#x2028; Results: MCF-7 cells overexpressing FL or LMW cyclin E had a higher level of P- SMAD 3 as compared to the parental and vector only control cells. Total SMAD 3 levels were similar in all cells examined. Transfection of WT Smad 3 elicited an increase in reporter activity in parental and vector control cells, while both FL and LMW forms of cyclin E overexpressing cells were resistant to WT Smad 3 reporter induction. In parental and vector control MCF-7 cells, expression of the Smad 3 Thr178 mutation resulted in a three-fold induction in Smad 3 reporter activity as compared to the WT Smad 3 response. In FL and LMW cyclin E overexpressing cells, induction by the Thr178 mutation was more pronounced, as seen by a 5-fold and 8-fold induction of reporter activity in FL and LMW cyclin E cells, respectively. Transient transfection of the Thr8 mutation failed to evoke a reporter response in any of the panel of MCF-7 cells. MCF-7, FL, and LMW cyclin E cells demonstrated the greatest fold induction of Smad 3 reporter activity when transfected with CDK 2 siRNA.&#x2028; Conclusions: Overexpression of cyclin E may circumvent the growth inhibitory effects of Smad 3 signal transduction through the modulation of Smad 3 function. Specifically, this work indicates that cyclin E mediates Smad 3 inhibition in a CDK 2 dependent fashion in breast cancer cells.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5033.
Induction of cell cycle arrest and apoptosis in breast cancer cells by troglitazone, a peroxisome proliferator-activated receptor-gamma (PPAR-γ) ligand.
Authors: Youn H , Lee B , Jung S , Park M , Yoon J , Lee J , Choi M 
Published: December 2010

Abstract
Abstract #5034
Background: Peroxisome proliferator-activated receptor-gamma(PPAR-γ) ligands inhibit cell proliferation and induce apoptosis in cancer cells. And, it is reported that PPAR-γ ligands could serve as negative regulators of breast cancer development and progression, but their mechanism is still unknown. Here we wished to determine whether the PPAR-γ ligand induces cell cycle arrest and apoptosis of MDA-MB-231(ERα-negative) and MCF-7(ERα-positive) breast cancer cell.&#x2028; Methods: The effect of PPAR-γ ligands on the cell viability of breast cancer cells was determined using mitochondrial tetrazolium(MTT) assay. The cell cycle distribution and apoptosis induction were evaluated by using the flow cytometry. The expression of apoptosis-related proteins were measured with Western blot analysis. Statistical analysis was performed using Student's t test, and p&lt;0.05 was considered significant.&#x2028; Results: The treatment of MDA-MB-231 cell with PPAR-γ ligand, troglitazone was shown to induce cell cycle G1 arrest and induction of apoptosis. Moreover, troglitazone treatment, applied in a dose-dependent manner, caused a marked decrease in phosphorylated retinoblastoma(pRb), cyclin D1, D2, D3, cyclin dependent kinase(Cdk) 2, 4, and 6 expression as well as a significant increase in Cdk inhibitor, p21 and p27. Troglitazone showed antiproliferative effect on MCF-7 cell with tamoxifen, respectively and synergically. Troglitazone and tamoxifen could induce G1 arrest and apoptosis of MCF-7 cell, through upregulation of Bax and downregulation of Bcl-2 and cyclin D1.&#x2028; Conclusion: PPAR-γ ligand, troglitazone induces cell cycle arrest and apoptosis of MDA-MB-231 cell and increases the sensitivity of anti-hormonal therapy in MCF-7 cell. These results suggest that troglitazone has anticancer effect on both ERα-negative and positive breast cancer cells.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5034.
Prevention of primary and contralateral breast cancer by risk reducing salpingo-oophorectomy in high risk women with a BRCA1/2 mutation or familial predisposition.
Authors: Heemskerk-Gerritsen BA , Hooning MJ , Collee M , Tilanus-Linthorst MM , Bartels CC , van den Ouweland A , Ansink A , Seynaeve C 
Published: December 2010

Abstract
Abstract #504
Introduction: Risk reducing salpingo-oophorectomy (RRSO) has become an important preventive option against ovarian/fallopian tube cancer, while it is also risk reducing regarding breast cancer (BC) in women with a BRCA1 or BRCA2 mutation. Recently a difference in efficacy of RRSO concerning the occurrence of BC has been suggested for BRCA1 and BRCA2 mutation carriers, while no data are available on the protective effect of RRSO in women from a hereditary breast/ovarian cancer (HBOC) family without a proven BRCA mutation. Further, it is not known whether the BC risk reduction after RRSO equally applies to primary (PBC) and contralateral BC (CBC).&#x2028; Methods: The occurrence of PBC and CBC was prospectively studied in 639 women ≥ 30 years (330 BRCA1 mutation carriers, 117 BRCA2 mutation carriers, and 192 HBOC women) who were enrolled in a breast/ovarian cancer surveillance program at the Rotterdam Family Cancer Clinic. Women with a history of BC without recurrent disease were eligible if BC treatment did not involve adjuvant systemic therapy. Demographics and follow-up (FU) information up to December 31, 2007, were extracted from the medical records. Ninety-eight BRCA1 mutation carriers, 33 BRCA2 mutation carriers and 15 HBOC women eventually underwent RRSO. Results were analyzed separately for women being at risk for PBC (n=489) and for CBC (n=164). The effect of RRSO on the incidence of BC was analyzed using a Cox proportional hazards model in which RRSO was modeled as a time-dependent covariate.&#x2028; Results: During a mean FU of 5.5 years, RRSO was associated with a reduction in BC risk of 48% (HR=0.52; 95% CI, 0.26-1.02; p=0.058) versus 81% (HR=0.19; 95% CI, 0.02-1.95; p=0.161) in BRCA1 and BRCA2 mutation carriers, respectively. In BRCA1 mutation carriers, PBC was reduced by 74% (HR=0.26; 95% CI, 0.08-0.89; p=0.032), and CBC by 29% (HR=0.71; 95% CI, 0.23-2.16; p=0.544). In BRCA2 mutation carriers no cases of PBC occurred after RRSO (HR=0,00; 95% CI, not estimable), while 1 CBC was diagnosed (HR=0.25; 95% CI, 0.01-5.62; p=0.379). In HBOC women no BC cases were observed after RRSO.&#x2028; Conclusions: RRSO is associated with a BC risk reduction in both BRCA1 and BRCA2 mutation carriers. The risk reductive effect is more pronounced regarding PBC as compared to CBC in BRCA1 mutation carriers. Although a similar trend is suggested for BRCA2 mutation carriers, larger numbers and longer FU are needed to draw firm conclusions.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 504.
Immune cells from pleural fluids of patients with inflammatory breast cancer are immunocompetent.
Authors: Cohen EN , Jimenez C , Gao H , Lee B , Li C , Krishnamurthy S , Woodward WA , Andreopoulou E , Bethancourt DL , Hortobagyi GN , Cristofanilli M , Reuben JM 
Published: December 2010

Abstract
Abstract #5044
Background: Malignant pleural effusion is a debilitating complication of metastatic breast cancer (MBC), including inflammatory breast cancer (IBC). The cellular components in pleural effusion are important indicators of pathologic process. Although pleural fluids from patients with IBC contain tumor cells and immune cells, little is known about the phenotype and function of the immune cell infiltrate. The objective of the study was to determine the immunophenotype and function of the cellular immune cells in pleural effusion of patients with pleural metastases.&#x2028; Methods and Patients: Pleural effusions from 9 women with MBC, 6 with IBC, median age 56 years (range 44-75 yrs), were submitted for pathological review. Mononuclear cells (MNC) were reacted with a cocktail of monoclonal antibodies to detect surface antigens by 8-parameter flow cytometry. In addition, MNC were activated through the T-cell receptor (TCR) with immobilized anti-CD3 and soluble anti-CD28 to assess intracytoplasmic syntheses of pro-inflammatory [interleukin (IL)-2, tumor necrosis factor (TNF)-a and interferon-gamma (IFN-γ)] and anti-inflammatory (IL-10) cytokines by TCR-activated CD4+ and CD8+ T cells. Data are presented as the mean (±SEM) of TCR-activated CD4+ and CD8+ T-cells that synthesized inflammatory and anti-inflammatory cytokines.&#x2028; Results: Upon pathological review, the mean (±SEM) number of tumor cells, MNC, and neutrophils were 20.2% ± 7.4% (range, 2%-72%), 62.4% ± 7.7% (range, 25%-96%), and 17.3% ± 5.9% (range, 0%-50.0%), respectively. The MNC consisted of 83.1% ± 8.2% CD3+ pan-T, 58.2% ± 7.0% CD4+ T, 24.2% ± 5.2% CD8+ T, 2.8% ± 0.9% CD19+ B, 4.0% ± 0.7% natural killer (NK), 3.4% ± 1.8% NKT, and 0.9% ± 0.2% dendritic cells, respectively. The CD4/CD8 T-cell ratio was 3.62 ± 0.78. The percentages of CD4+ and CD8+ T cells that synthesized inflammatory and anti-inflammatory cytokines following activation through the TCR-activated are listed in Table 1.&#x2028; &#x2028; Conclusion: There were no statistically significant differences between pleural fluids of IBC and non-IBC subjects with respect to tumor cell involvement, MNC phenotype, or T-cell function. Moreover, the function of pleural fluid T cells of MBC patients were similar to that of normal peripheral blood T cells (historical data, not shown). These data suggest that T-cells in the pleural fluids of patients with MBC are immune competent and can be activated through the TCR to synthesize pro- and anti-inflammatory cytokines.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5044.
A new paradigm for African American breast cancer involving stem cell differentiation.
Authors: Latimer JJ , Lalanne NA , Myers NT , Courtney AB , Mock L , Grant SG 
Published: December 2010

Abstract
Abstract #5051
Background: African American (AA) women have a higher incidence of breast cancer (BC) than white women before age 40, and are more likely to die from BC at every age. These differences are often attributed to socio-economic factors, however, there may be intrinsic biological differences in the breast tissue of AA and white women. For example, triple negative breast tumors are more prevalent in AA women than in non-Hispanic white women, although neither population is monolithic in its breast cancer phenotypes. We hypothesis that intrinsic biological differences between women of AA and European-white ancestry might be revealed by differences in growth and differentiation in vitro, reflecting underlying differences in the proportions and/or potency of breast epithelial stem cells.&#x2028; Methods: Our laboratory has developed a novel tissue engineering system for Human Mammary Epithelial Cells (HMEC), both normal and malignant. This system allows for long-term establishment of non-diseased primary cultures that begin as 3-dimensional attached “epispheres”, that are structures made up of 40-100 epithelial cells. These non-diseased epispheres subsequently differentiate into functional, organotypic branching ducts and lobules that demonstrate ESA and CK -18,-19 staining, lumen, polarized nuclei, desmosomes, microvilli on apical surfaces and casein secretion.&#x2028; Results: We have established primary HMEC cultures from 36/36 breast reduction mammoplasty tissues, including 9 AA donors matched in socioeconomic status. We have found: 1) The more children a woman had, the less likely her breast tissue was to form ductal structures in vitro. This finding is consistent with the idea that lactational differentiation decreases the number of pluripotent stem cells in the breast. 2) Pre-menopausal breast tissue was more likely to form ductal structures than post-menopausal tissue. 3) Race was a modifying factor in the ability to form ductal architecture in culture, significantly affecting the rate of differentiation. Putative mammary stem cells were analyzed by flow cytometry (CD44+, CD24-, MUC1-, α6 integrin+) and cloning. Stem cell proportions varied widely (range 0.5-15%), with no obvious association with race.&#x2028; Discussion: One interpretation of the hormonal risk factors for BC involves the susceptibility of undifferentiated breast tissue to transformation. In other systems, especially the gonads, interference with terminal differentiation is also associated with susceptibility to transformation. If AA breast tissue has more robust differentiation potential than that of white women, it may also contain stem cells more likely to transform into an aggressive tumor type.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5051.