bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting novel microtentacles to reduce the adhesion of circulating tumor cells to blood vessel walls.
Authors: Whipple-Bettes RA , Balzer EM , Yoon JR , Cho EH , Matrone MA , Martin SS 
Published: December 2010

Abstract
Abstract #2044
Detached human breast tumor cell lines produce novel protrusions of the plasma membrane that promote reattachment of tumor cells to each other and extracellular matrix. We have termed these protrusions “microtentacles” due to clear mechanistic distinctions from either actin-based filopodia/invadopodia or tubulin-based cilia.&#x2028; &#x2028; Compelling in vivo studies show that a mechanism consistent with microtentacles is responsible for the attachment of circulating tumor cells to blood vessel walls. Human breast tumor cell lines that display a disorganized actin cortex and hallmarks of epithelial-to-mesenchymal transition produce higher frequencies and greater extension of microtentacles. Conversely, some current and proposed tumor drugs that target the cytoskeleton actually increase microtentacles. While many cancer therapies are directed against the process of cell division, it remains important to consider the effects of these drugs on the invasive characteristics of tumor cells. Our current research has determined that microtentacles are composed of coordinated filaments of vimentin and detyrosinated tubulin. Previous studies indicate that elevated levels of vimentin and detyrosinated tubulin in tumors predict poor patient outcome, although the specific mechanisms remain unclear. Time-lapse video microscopy also indicates that the rapid extension and dynamic motion of microtentacles relies on kinesin activation. Again, our results suggest that the kinesin responsible is a conventional kinesin, rather than the mitotic kinesins commonly targeted by cancer therapies. We propose that the independent roles of vimentin, detyrosinated tubulin and kinesin in tumor metastasis could result from their interdependent contributions to microtentacle formation. Since large epithelial tumor cells are often fragmented in narrow capillaries, we are targeting microtentacles to promote the fragmentation of circulating tumor cells in the capillaries of the lung. We are currently using time-lapse confocal microscopy of tumor cells circulating in intact blood vessels and whole-animal optical imaging to define therapies that reduce microtentacle-dependent adhesion of circulating breast tumor cells during metastasis.&#x2028; &#x2028; This image shows circulating tumor cells (red) adhering to the walls of an intact blood vessel (green).
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2044.
A transcriptional regulatory mechanism in invasive breast cancer; role of AIB-1 and PEA-3.
Authors: Al-Hilli Z , Redmond AM , Hill AD , Young LS 
Published: December 2010

Abstract
Abstract #2056
The invasive potential of cancer cells is mediated in part through the action of matrix metalloproteinases (MMP), which degrade the extra-cellular membrane and promote metastasis. The transcription regulatory region of MMP genes contains binding sites for Ets transcription factors. While the underlying mechanism of MMP regulation is unclear, the Ets transcription factor PEA3 has been shown to be involved. We have previously reported that expression of PEA-3 is associated with the co-activator AIB-1, HER2 positivity, metastasis, and reduced disease free survival. Furthermore, an AIB-1 knockout mouse exhibited PEA-3 dependant MMP production, resulting in reduced metastatic potential. In this study we combined translational and molecular approaches to study the role of PEA-3 and AIB-1 in the transcriptional regulation of MMPs in breast cancer.&#x2028; The expression of PEA-3, AIB-1, and MMP-9 was assessed in a tissue microarray of breast cancer patients (n=560). Strong associations were observed between both PEA-3, AIB-1, and their target gene MMP-9 (p= 0.007, p= 0.0016 respectively). In HER2 positive patients PEA-3, but not AIB-1 or MMP-9, was associated with reduced disease free survival (p&lt;0.0044, p= 0.4491, and p= 0.0666 respectively). A stable breast cancer cell line over-expressing PEA-3 was created. Upregulation of the growth factor pathway in this cell line resulted in the increased expression of MMP-9. Furthermore, silencing AIB-1 resulted in reduced MMP-9 production in breast cancer cells in vitro.&#x2028; Ets transcription factors are important mediators of breast tumour invasion. This data supports a role for the transcription factor PEA-3 and its co-activator AIB-1 in the production of MMPs in breast cancer.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2056.
EGF-dependent enhancement of invasive ability in squamous cell carcinoma of the breast.
Authors: Kimura F , Iwaya K , Kawaguchi T , Yamada K , Ogata A , Kaise H , Komatsu S , Nakamura Y , Ueda N , Mukai K , Kohno N 
Published: December 2010

Abstract
Abstract #2061
The factors promoting the clinical aggressiveness of squamous cell carcinoma of the breast is still not well understood. In order to examine whether the mechanism of cell motility is involved in the aggressive behavior of squamous cell carcinoma, we established a squamous cell carcinoma cell line from breast primary (HBC 9) and assayed its invasiveness. The number of HBC9 cells migrating through matrigel was significantly increased by stimulation with epidermal growth factor (EGF). This enhancement of cell motility was distinctly detected in comparison with four ductal carcinoma cell lines (MCF7, BT474, HBC5, and MDA-MB-231). EGF stimulation also induced surface protrusions of HBC9 cells. To examine the interaction between Arp2/3 complex and N-WASP which is the final intracellular signal of invadopodia formation that is downstream from EGF stimulation, we observed surface protrusion of HBC 9 by laser microscopy. Co-localization of Arp2 and N-WASP was detected with actin polymerization visualized by phalloidin. Furthermore, the localization of cortactin was found to agree with that of N-WASP and Arp2/3 complex in the invadopodia. The cortactin gene copy number was evaluated using real time PCR by comparison between 12 DNA samples extracted from primary mammary squamous cell cancer tissue and those from corresponding normal breast tissue. A higher level of cortactin was detected in cancer tissue than in normal tissue in 10 cases (83%). At a protein level, intense immunostaining of cortactin was detected in all 12 cases, among which expression was confirmed in 2 cases by Western blot analysis. These results suggest that one of the causes of clinical aggressiveness of squamous cell carcinoma of the breast is the enhancement of cell motility by invadopodial formation. Overexpression of cortactin may result in the formation of invadopodium.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2061.
Quantitative gene expression analysis using Oncotype DX in ductal carcinoma in situ that is adjacent to invasive ductal carcinoma.
Authors: Baehner FL , Sangli C , Millward C , Cherbavaz D , Goddard A , Shak S 
Published: December 2010

Abstract
Abstract #2066
Background: It is common for ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) to be observed adjacent to each other in the same tissue section. To explore the quantitative biology of DCIS, we measured ER, PR, HER2, proliferation gene expression and Recurrence Score (RS) using Oncotype DX in DCIS adjacent to IDC.&#x2028; Material and Methods: Thirty cases of ER positive tumors were identified with adjacent DCIS and IDC of similar nuclear grade. Manual microdissection was performed to separate DCIS and IDC tissues from each case. Using the Oncotype DX assay, standardized quantitative expression of 16 individual cancer related genes including ER, PR and HER2 and the proliferation index (the average of the 5 proliferations genes) was measured on a scale from 0 to 15 (relative to reference genes), where a one unit increment is associated with a 2-fold change in expression. The RS was also calculated. Descriptive statistics and Pearson's R correlation coefficients were determined to compare DCIS to IDC.&#x2028; Results: 27 of the 30 samples had sufficient RNA for analyses of the DCIS component (8 with low grade, 10 with intermediate grade, and 9 with high grade DCIS). The correlation coefficients for the comparison of DCIS and adjacent IDC for quantitative ER, PR, HER2, and the proliferation index was 0.78, 0.73, 0.76, and 0.83, respectively. For ER, IDC and DCIS measures were within 2 units for all 27 cases and within 1 unit for 22/27 (81%). For PR, IDC and DCIS measures were within 2 units for 25 of 27 cases and within 1 unit for 19/27 (70%). For HER2, IDC and DCIS measures were within 2 units for 26 of 27 cases and within 1 unit for 24/27 (89%). For proliferation index, IDC and DCIS measures were within 2 units for all 27 cases and within 1 unit for 26/27 (96%). There was a wide range of RS in DCIS (mean 13.9; range 0 to 56). Although for RS there was a significant correlation between DCIS and IDC (R=0.84), on average the RS was 4 units lower for DCIS.&#x2028; Discussion: Quantitative RT-PCR analysis of DCIS is technically feasible and sufficient RNA for Oncotype DX analysis is obtained in 90% of cases. As has been described previously for invasive breast cancer, there is considerable expression heterogeneity in DCIS between patients. DCIS adjacent to invasive breast cancer demonstrates very similar, but not identical, quantitative biology for the genes in the assay. Further DCIS studies to evaluate Oncotype DX and recurrence rates would be of great interest.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2066.
Bone-specific alkaline phosphatase (BSAP) and serum N-telopeptide (sNTX) as predictors of bone loss in postmenopausal women with early breast cancer receiving letrozole as adjuvant therapy: a 5-year study (Z-FAST).
Authors: Brufsky A , Harker G , Beck T , Carroll R , Warsi G , Jin L , Argonza-Aviles E , Ericson S 
Published: December 2010

Abstract
Abstract #2067
Background: Changes in biochemical markers of bone turnover are expected to be predictors of long-term changes in bone mineral density (BMD). We studied the potential of bone-specific alkaline phosphatase (BSAP) and serum N-telopeptide (sNTX) at 6 and 12 months (mo) in predicting BMD of the lumber spine (LS) and total hip (TH) at 48 mo in Z-FAST study.&#x2028; Methods: Postmenopausal women with ER+ and/or PR+ early stage breast cancer were randomized 1:1 into Up-front (U) or Delayed (D) arms (N=301 in each). Patients (pts) in U received zoledronic acid (ZOL) 4 mg IV q6 mo starting at baseline; pts in D started ZOL q6 mo when their LS or TH T-score fell below -2. All pts started Letrozole 2.5 mg/day for 5 years. Levels of BSAP and sNTX were assessed at screening and every 6 mo on a subset of pts. LS and TH BMD were assessed by dual-energy x-ray absorptiometry (DEXA) at screening, 6 mo and every 12 mo thereafter for all available pts. This is an exploratory analysis to investigate the correlation between short-term % changes from baseline in BSAP and sNTX and long-term % changes from baseline in BMD. Pearson correlations and Receiver Operating Characteristics (ROC) analysis were performed. We report the results based on 48 mo data.&#x2028; Results: The Pearson correlations between % change in BSAP at 6 mo and LS BMD at 48 mo were -0.13 [(N=38, not significant (NS)] and -0.17 (N=39, NS) in arms U and D respectively. The correlations between % change in BSAP at 12 mo and LS BMD at 48 mo were -0.15 (N=39, NS) in U and -0.43 (N=38, p=0.0077) in D. Very similar correlations were obtained for % change in sNTX at 6, 12 mo and LS BMD at 48 mo in D, but in U the correlations were smaller and positive. The correlations between % change of both markers at 6, 12 mo and TH BMD at 48 mo were all NS at 0.05. ROC analysis showed that at best performance (maximum of sensitivity plus specificity), for any increase in sNTX at 12 mo, the sensitivity (SE), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) to predict LS BMD loss at 48 mo was 74%, 77%, 85% and 63% in D. For BSAP change of 3%, the four ROC parameters were 71%, 71%, 81% and 59% respectively. For prediction of TH BMD loss, the SE, SP and NPV were lower for both markers with higher PPV. In U, the SE, SP and PPV were all much lower with very high NPV for both markers and BMD loss.&#x2028; Conclusion: Interim results of Z-FAST data show that short-term changes in BSAP and sNTX have the potential for predicting long-term bone loss. The weaker correlations in U also suggest that zoledronic acid may have a direct preventive effect on bone loss independent of changes in the markers.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2067.
Fluorescence in situ hybridization (FISH) technique by reference centers for HER2 status determination in metastatic breast cancer: quality assurance results of FISH 2002 study.
Authors: Penault-Llorca F , Lannes B , Vincent-Salomon A , MacGrogan G , Vilain M , Fermeaux V , Treilleux I , Blanc-Fournier C , Leroux A , Antoine M , Ettore F , Charpin C , Jacquemier J , Escourrou G , Lacroix-Triki M , Gosset P , Sagan C , Arnould L , Couturier J 
Published: December 2010

Abstract
Abstract #2069
Background: Concordance between IHC and FISH in local and regional centers was evaluated in the FISH 2002 Study. Primary metastatic breast cancer (MBC) tumor samples were tested by IHC for HER2 status according to in-house techniques at 18 regional (n=871) and 81 peripheral (n=670) centers in France between 2002 and 2006. All samples were tested by FISH at regional centers [Penault-Llorca et al. SABCS 2006]. This second part of the study comprises quality assurance evaluation of FISH techniques.&#x2028; Methods: 464 cases were selected according to the protocol and were either re-assessed using archived pictures or retested by 3 reference centers (Clermont-Ferrand, Strasbourg, Paris-Curie). 246 samples were re-assessed and 166 were retested.&#x2028; Results:Global discordance combining re-assessment and retesting results was 10.7% in the first half of the study and 5.1% in the second part (p=0.041).&#x2028; &#x2028; Discussion: Concordance was excellent for negative (100%) and IHC 2+ cases (97%). Among low amplification cases (≥6 and &lt;8 copies), 70% were concordant: discordant cases were mainly interpretation pitfalls of polysomic cases, emphasizing the importance of double-stained FISH analyses. These quality assurance results demonstrate that overall concordance and quality of HER2 status determined by FISH has improved significantly over time. This improvement is likely to be related to introduction of rigorous quality control procedures, training, and education, all of which should be continued.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2069.
Phosphorylated c-Src predicts clinical outcome in triple negative breast cancers.
Authors: Elsberger B , Tovey SM , Tan BA , Brown SB , Brunton VG , Mallon EA , Cooke TG , Edwards J 
Published: December 2010

Abstract
Abstract #2076
Background: Recent in vitro evidence suggests that c-Src inhibitors may be effective in inhibiting growth of basal-type/"triple-negative" cell lines. However there is little evidence to support this in clinical specimens. Development of treatment for women with these aggressive ER, PR and HER2 negative cancers is particularly important with the lack of current effective therapies. Activation of Src family members are associated with phosphorylation at two sites Y419: the classical activation site and Y215, known to induce a 50-fold increase in activation.&#x2028; The aim of the current study is to assess if expression levels and/or activation of Src kinase is associated with clinical outcome measures in ER and HER2 negative human breast cancers.&#x2028; Methods: Tissue microarrays consisting of 172 ER and HER2 negative breast cancer tumors were constructed. Median clinical follow up was 5.9 years with 64 deaths attributed to breast cancer. Immunohistochemistry was performed using antibodies to c-Src, pSrc419 and pSrc215 (antibodies to Y419 and Y215 will detect phosphorylation of c-Src and other family members). Expression was assessed by two independent scorers. All statistical calculations were performed using SPSS 15, including Kaplan-Meier life table analysis with log rank testing of differences in breast cancer related survival.&#x2028; Results: Membrane expression of c-Src, pSrc419 and pSrc215 was rarely observed and was not associated with outcome. However cytoplasmic expression was frequently observed. There was a trend towards reduced survival seen with c-Src and pSrc419 overexpression. In direct contrast pSrc215 was significantly associated with increased disease specific survival and was independent on multivariate analysis.&#x2028; We then hypothesised that activation of c-Src via Y419 but not Y215 was driving the observed association with outcome. Indeed c-Src positive, pSrc419 positive and pSrc215 negative patients have a 25% 5yr survival rate, in comparison all other patients in the cohort had a 72% 5yr survival rate.&#x2028; &#x2028; Discussion: These results support the in vitro studies suggesting Src family members are associated with progression of triple negative breast cancers. If confirmed in a larger cohort, these results suggest that Src family members may offer a therapeutic target for triple negative cancers. However care must be taken to appropriately select patients with phosphorylation only at Y419 and not Y215. The reason why Y215 phosphorylation is associated with improved outcome requires further investigation.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2076.
Time distributions and prognosis factors of breast cancer recurrence in different sites after operation.
Authors: Zhou L , Yin W , Lu J , Di G , Wu J , Shen K , Shen Z , Shao Z 
Published: December 2010

Abstract
Abstract #2080
Objective Our study aims to analyze the breast caner patients with different sites of recurrence, and to have knowledge of the regular patterns and influence factors of metastastic breast cancer.&#x2028; Methods We performed a retrospective study of 210 female breast cancer patients who were continuously followed up in our hospital from January 2005 to May 2007 and had complete data of metastasis. The clinical and biological characteristics were compared between the patients with organ metastases (lung, liver, brain) and non-viscera metastases (lymph node, bone).&#x2028; Results Cox regression analysis-hazard function showed that patients with elder age (RR=0.927 95%CI 0.877∼0.981, P=0.008) and HER-2 negative(RR=0.253 95%CI 0.076∼0.836, P=0.024)tended to develop non-viscera metastases, while patients with larger tumors (RR=3.832 95%CI 1.073∼13.687, P=0.039) tended to develop organ metastasesp.&#x2028; &#x2028; When stratified by the prognostic factors, the annual risk hazard curve of the patients with high risk of recurrence had double peaks at the second year and the ninth years after mastectomy respectively, and it was significantly higher than those of the patients with middle and low risk.[figure1]Annual risk hazard curves of both local relapse and metastasis also showed a double-peaked pattern. The curves for lymph node, bone, lung and liver metastasis showed a similar pattern, but all the peaks of curve emerged later than that of the local relapse.&#x2028; &#x2028; Conclusion Patients with different clinical and biological characteristics may develop recurrence in different sites. There are certain regular patterns of time distribution for different metastasis sites after mastectomy. The local recurrence may be the foreboding for the metastasis and we should pay attention to the local relapse signs especially in the patients with high risk.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2080.
Noggin, impact on in vitro cell growth, expression pattern and the prognostic value in breast cancer.
Authors: Ye L , Mansel RE , Jiang WG 
Published: December 2010

Abstract
Abstract #2084
Background: Noggin, a BMP (bone morphogenetic proteins) antagonist, has been demonstrated to play critical roles in regulation of biological functions of BMPs, members of the TGF-β superfamily and known regulators of bone metastasis in solid tumours including breast cancer. We have recently reported that the presence of endogenous noggin in cancer cells is a pivotal factor that governs the cells response to stimulation by bone morphogenetic protein, for example BMP-7. However, the role of noggin in breast cancer remains unclear. We presently investigated the role of Noggin in breast cancer cells and the pattern of expression in clinical breast tumours.&#x2028; Methods: The expression of Noggin mRNA and presence of Noggin protein was examined in a cohort of human breast cancer specimens (normal, n=33; cancer, n=112), using both quantitative real time PCR and immunohitochemical staining, respectively. The full-length human Noggin was cloned into a mammalian expression plasmid vector (pEF/His TOPO TA plasmid vector). The constructed Noggin expressing constructs were then transfected into breast cancer cells (MDA-MB-231). The expression of Noggin in the transfected cells was determined using RT-PCR and western blot. The effect on growth of MDA-MB-231 cells by Noggin was investigated using an in vitro growth assay.&#x2028; Results: Immunohistochemical staining revealed that Noggin protein was abundant in the cytoplasmic region of normal epithelial cells. Reduced staining of Noggin in breast cancer cells was seen when compared to normal breast tissue. Quantitative real time PCR further demonstrated that the decreasing Noggin expression correlated to poor prognosis, in that patients who remained disease-free showed a higher levels of the transcript (11113±3612) compared with those who had a poor clinical outcome (with metastasis and mortality, 6420±2241). Kaplan-Meier survival analysis showed that patients with high Noggin level had a longer survival (134.4 (121.3-147.5 - 95%CI) months) that those with low levels (102.8 (80.8-124.8) months). The absorbance of MDA-MB-231 Noggin over-expressing (MDA-MB-231Nogginexp) cells of Day 5 is 1.07±0.13, p&lt;0.01 vs both MDA-MB-231 wild type (MDA-MB-231WT, 1.54±0.08) and empty vector control (MDA-MB-231pEF/His, 1.72±0.17).&#x2028; Conclusions: Noggin inhibits the growth of breast cancer cells and has a pattern of expression in that decreased Noggin expression correlates to poor prognosis of breast cancer patients.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2084.