bluuebunny/crossref_metadata_2025_split huggingface dataset:
P5-01-07: Identification of SORBS2 as a Candidate Marker To Predict Metastatic Relapse in Breast Cancer.
Authors: Alsafadi S , Scott V , Pautier P , Goubar A , Lazar V , Dessen P , Lacroix L , Duvillard P , Morice P , André F , Delaloge S 
Published: June 2012

Abstract
Background
Elucidation of promising cancer biomarkers from gene expression data can provide important insight into the relationship between signaling networks and cancer. SORBS2, sorbin and SH3 domain containing 2, is a multi-adapter protein involved in signal transduction associated to the cytoskeleton and was reported to be strongly repressed in pancreatic and cervical cancers.
Methods: With the purpose of identifying genes involved in metastatic process, we compared gene expression profiling of 19 invasive ovarian cancers and 24 borderline tumors. Prognostic value of the selected genes was then tested in a gene expression array database that includes 1659 patients with early breast cancer (Gyorffy B et al. 2010). Upon isolation of SORBS2 as a predictor, its involvement in cell migration and tumor progression was investigated in vitro. Small interfering RNA targeting SORBS2 was used to downregulate its expression in T47D and Hela, two cell lines overexpressing SORBS2. Functional effect of siRNA-induced knockdown of SORBS2 on cell viability was determined by WST-1 assay and Trypan Blue exclusion test. Effect on cell migration was evaluated by wound-healing and transwell assays. Western blot analyses were also performed to examine the expressions of proteins involved in cell survival, death and migration.
Results: High-throughput analyses of genes that are differentially expressed between borderline ovarian tumors and invasive carcinoma demonstrated that SORBS2 is significantly downregulated in invasive carcinoma (FDR&lt;10e-10). Moreover, SORBS2 loss was a strong predictor of relapse in a gene expression array dataset of 1659 patients presenting an early breast cancer (HR=0.62, 0.52−0.72, p=6.4e-10). In vitro studies showed that SORBS2 specific downregulation stimulated cell proliferation, by an average of 40%, in T47D and Hela cell lines. SORBS2 knockdown had no effect on either cell migration rate or protein expressions of p-FAK and p-Paxillin, two focal adhesion proteins which are critical to cell adhesion and migration. No significant variations in protein expressions of Akt/mTOR pro-survival or caspases activation pathways were observed. Gene expression profiling before and after SORBS2 dowregulation is ongoing in the two cell lines.
Conclusion: This study is the first to provide evidence for an antiproliferative activity of SORBS2 with no effect on cell migration in breast cancer cells. Our clinical and in vitro data suggest that SORBS2 is a candidate marker to predict relapse in patients with early breast cancer. Molecular mechanisms mediating the antiproliferative effect of SORBS2 are currently being investigated.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-07.
P5-01-15: The Functional Role of the Estrogen-Regulated Gene GREB1: Characterization of a Novel GREB1 Knockout Mouse Model.
Authors: Hnatyszyn HJ , Rodriguez C , Herbert L , Olson R , Lippman ME 
Published: June 2012

Abstract
Background: Gene regulated in breast cancer 1 (GREB1) was initially discovered in breast cancers as an estrogen-regulated gene that mediates estrogen-stimulated cell proliferation and is a candidate clinical marker for response to endocrine therapy. However, little is known of the functional role of GREB1 protein in normal breast tissue or breast cancers.
Methods: To address this unknown role, our laboratory designed and created a novel Greb1 Knockout Mouse model (C57/bl MEL Greb1 KO). This constitutive model results in the loss of Greb1 mRNA and protein expression in cells where expression of Cre recombinase promotes the cleavage of exon 1 and intron 1 of the gene encoding Greb1. ROSA26 Cre C57/b1 MEL Greb1 KO mice heterozygous for the floxed Greb1 allele were crossed to generate experimental litters. Initial experiments were designed to evaluate if the complete loss of Greb1 expression in offspring homozygous for the floxed Greb1 allele was lethal during gestation. Experimental litters were tail clipped and genotyped using gDNA and genotype-specific PCR.
Results: Offspring homozygous for the floxed Greb1 allele were identified in expected Mendelian ratios with wild type and heterozygous siblings. Loss of Greb1 expression was confirmed using RT-PCR, in situ hybridization and immunoblotting. Loss of both Greb1 alleles was not observed to be lethal during gestation for either male or female pups. Preliminary gross observation of these homozygous KO mice revealed no overt anatomical differences, however, they were 25–30% smaller than their heterozygous and wild-type siblings. Breeding experiments are underway to determine the fertility of crossbred Greb1 homozygous KO mice. Imaging experiments and necropsy with histochemical analysis of tissues will reveal any alteration in architecture and function. These findings will be summarized in this presentation.
Discussion: As GREB1 has been identified as an estrogen-regulated gene involved in breast cancer cell proliferation and a potential target for new therapeutic strategies, it is important to understand the contribution of GREB1 to the differentiation, development and function of normal tissues as well as in breast cancers. Characterization of this novel Greb1 KO mouse model will provide answers to these functional questions surrounding GREB1.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-15.
P5-01-17: HER2 Amplification in Primary Tumor: A Potential Marker for Presence of Circulating Tumor Cells in Inflammatory Breast Cancer Patients?
Authors: Bhattacharyya A , Krishnamurthy S , Lodhi A , Hall C , Anderson A , Jackson S , Ueno N , Bedrosian I , Kuerer H , Lucci A 
Published: June 2012

Abstract
Background: Inflammatory breast cancer (IBC) is a rare but aggressive form of invasive breast cancer accounting for 3–6% of all cases and have higher rates of distant recurrence. Circulating tumor cells (CTCs) are known to predict outcome in metastatic breast cancer (BC) patients, but little is known about their prognostic significance in non-metastatic BC. We hypothesized that CTCs can be identified in patients with IBCs and may correlate with primary tumor characteristics. Methods: All patients had blood samples collected at the time of primary surgery. CTCs (per 7.5 ml blood) were detected using the Cell Search™ system (Veridex) and were defined as nucleated cells lacking CD45 but expressing cytokeratins (CK) 8, 18, or 19. The presence of ≥ 1 epithelial cells meeting morphologic criteria for malignancy was considered a positive result. Statistical analyses employed Chi square and Fisher's exact tests using STATA IC 11. Results: We prospectively evaluated 41 IBC patients enrolled in an IRB approved protocol undergoing surgery for stage I-III breast cancer. Median follow-up was 30 months. Mean age was 52 years. Thirty five patients (94%) had positive lymph nodes (LNs) at presentation, 30 (75%) had high-grade tumors and 20 (53%) had lymphovascular invasion. Eleven patients (28%) were ER positive, 11 (27%) were PR positive and 18 (44%) were HER2 positive. IBCs were more likely to be high grade (P&lt;0.0001), ER negative (P&lt;0.0001), PR negative (P&lt;0.0001), HER2 positive (P&lt;0.0001), High Ki-67 (P= 0.005) and had a BMI of more than 25kg/m2 (P=0.04). Eleven (27%) patients were CTC positive. CTCs were more likely be found in HER2 positive (8/18; 44%) vs. HER2 negative primary tumors (3/20; 15%) [OR= 4.53; 95% C.I. = 1.02−19.52; P= 0.04]. We found no statistically significant correlations between primary tumor characteristics (ER, PR, LNs, high grade) and presence of CTCs. Conclusions: About a quarter of IBC patients had CTCs at the time of primary surgery. In these patients HER2 overexpression predicted the presence of CTCs. Studies with longer follow-ups is needed to determine if CTCs predicted survival.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-17.
P5-06-06: The RNA Binding Protein FUS Is a Potential Marker for Breast Cancer Progression and Therapy Response.
Authors: Brooke GN , Bevan CL , Rudraraju B , Palmieri C 
Published: June 2012

Abstract
Background
The RNA binding protein FUS is a multifunctional protein that has recently been demonstrated to be directed to target genes via non-coding RNA, where it represses transcription via disruption of the transcriptional complex. We have shown that FUS is an important regulator of prostate cancer cell growth; FUS over-expression causes G1 arrest, promotes apoptosis and blocks proliferation in vitro and in vivo, whereas knock-down increases proliferation. This regulation appears to be, at least in part, via direct regulation of cyclin D1. Further, analysis of patient samples demonstrated that FUS levels are inversely correlated with Gleason grade and directly correlated with survival and the presence of bone metastases. We hypothesised that FUS is a predictive and/or prognostic marker for hormone-dependent cancers.
Materials and Methods: FUS expression in breast cancer was analysed in silico and using Western blotting and immunohistochemistry on a tumour microarray. FUS levels were knocked down by siRNA in MCF-7 cells and chemosensitivity analysed using caspase 3/7 assays.
Results: Analysis of the Finak dataset (2008) demonstrated that FUS expression is significantly lower in breast cancer compared to normal breast. Immunohistochemistry revealed that FUS expression is also inversely correlated with grade. Importantly, FUS expression was also correlated with survival (Kaplan-Meier Plotter); significantly longer relapse free survival was seen in patients whose tumours had high levels of FUS. FUS expression also appears to correlate with therapy response since analysis of the Chang dataset (2003) revealed that FUS levels are significantly lower in patients resistant to Docetaxel and we found FUS levels to be lower in the tamoxifen resistant MCF-7 cell line (MCF-7 TAMR) compared to the parental line. We therefore hypothesised that loss of FUS expression may be driving therapy failure. In support of this, reducing FUS expression in the parental MCF7 line resulted in a significant reduction in docetaxel-induced Caspase 3/7 activity, suggesting that FUS is a regulator of chemosensitivity.
Discussion: FUS may be an important regulator of breast cancer progression since its levels are inversely correlated with grade and directly correlated with relapse free survival. Further, lower FUS expression is also correlated with reduced hormone therapy and chemo-sensitivity. Since knock-down of FUS reduced MCF-7 chemosensitivity, we believe that the observed changes in FUS expression are, in part, driving therapy failure and hence propose that FUS is a novel target and biomarker for breast cancer.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-06-06.
P5-07-06: Effect of Angiotensin-(1-7) and Angiotensin II on T47D Breast Cancer Cells in the Proliferation and cAMP Production.
Authors: da Silva IDCG , Correa-Noronha SSAA , Noronha SMR , Alecrim C , Shimuta SI , Nakaie CR , Gebrim LH , Nazario ACP 
Published: June 2012

Abstract
Angiotensin-(1-7) [Ang(1-7)] is a peptide hormone that produces opposite responses to those of the well-characterized peptide angiotensin II (Ang II). The main actions of Ang(1-7) and AII occur, respectively, via MAS and Angiotensin type 1 receptor (AT1R), respectively. Ang (1-7) has blood pressure and proliferative effects contrary to those of AngII, exhibits significant antiangiogenic activity and may be a novel therapeutic agent for lung cancer treatment targeting a specific AT(1-7) receptor (Soto-Pantoja et al., 2009). Here, our aim was to evaluate T47D breast cancer cell proliferation and cAMP production after treatment with Ang (1-7) and AngII.By means of the cyclic AMP (cAMP) competitive enzymeimmunoassay system we measured cAMP content of T47D cells. Cell proliferation was measured using incorporation of BrdU after Ang (1-7) and AII stimulation. Intracellular cAMP increasing drugs like Forskolin (FK) and isobutylmethylxanthine (IBMX) were also used to stimulate T47D cells.AII inhibited cAMP production (−39%) and A-(1-7) increased its formation (48%). Pattern of cAMP production was not altered when cells were previously incubated with FK. Results were inverted after IBMX incubation. In proliferation, opposite responses were observed for Ang II and Ang (1-7), that is, increasing (7.6 times) and decreasing (7.5 times) proliferation rates respectively. Also both peptides decreased proliferation when cells were stimulated by FK. Exactly the opposite happened after IBMX stimulation, both peptides increased proliferation. These results suggest that Ang (1-7) inhibits growth of T47D cells probably through inhibition of the via RAS/RAF/MAP quinase, what corroboraties previous findings. On the other hand, AngII can induce cell proliferation by an alternative via (CREB/growth factor). Therefore, Ang (1-7) seems to be not only a promising target to develop novel lung cancer therapeutic drugs, but it also might help to prevent the molecular changes that lead the normal breast epithelial cells to cancer initiation.
Financial Support: FAPESP.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-07-06.
P5-11-08: High Concordance of 5 HER2 In Situ Hybridization Methods with Abbott FISH.
Authors: Boers JE , Netjes C , Meeuwissen HC , Prinsen C , Bart J , van der Logt EMJ , Schuuring E 
Published: June 2012

Abstract
Background: HER2 in situ hybridization (ISH) has become a common test in breast cancer. Abbott FISH was used in most clinical studies showing the efficacy of anti-HER2 treatment in HER2 positive carcinomas. Only reports comparing one or two of newly developed ISH assays with Abbott FISH have been published previously. We conducted a comprehensive concordance study of 5 ISH methods with Abbott FISH in a large series of breast carcinomas.
Methods: Tissue Micro Arrays (TMA) were constructed by taking three 0.6 mm tissue cores from formalin-fixed/paraffin-embedded tissue-blocks from 402 primary breast carcinomas diagnosed in 2007 (supported by the Dutch Pathological Society). Up to 384 cases were analyzable in the TMA. ISH was performed after ample experience with 6 ISH assays. Scoring was performed by two independent observers without knowledge of the other ISH data according the ASCO-guidelines for HER2−testing. HER2 and chromosome 17 (Chr17) signals were counted separately, the HER2:Chr17 ratio was calculated and considered positive when the ratio was ≥ 2.0 In cases with a ratio was between 1.8 and 2.2, additional enumeration was performed. The discordant cases were reviewed and scores were reassigned on consensus of opinion. Concordance and Cohen's kappa score were calculated.

Conclusion: Concordance of 5 HER2 ISH assays with Abbott FISH were shown to be 98% or higher. Each of these tests might be used for a reliable assessment of HER2 amplification.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-11-08.
P5-11-12: Correlation of Ki67 Expression between Initial Biopsy and Surgical Specimen in Untreated Breast Cancer Patients. Does Menstrual Cycle Matter?
Authors: Torrejón D , Rubio I , Sanchez-Olle G , Peg V , Sansano I , Jimenez J , Gomez P , De la Peña L , Di Cosimo S 
Published: June 2012

Abstract
Background: The measure of proliferative index by Ki67 expression is being increasingly used to identify breast cancer subtype and to guide treatment decisions. Findings on initial tumor biopsy are in some cases, such as for primary systemic treatment, the only data available prior to treatment. In addition, results from window studies to develop novel anti-tumor agents depends on initial breast cancer biopsy findings. This study aims to compare the Ki67 expression on initial biopsy with matched primary breast cancer surgical specimens. In addtion, we aimed to explore the possible effect of hormonal levels — as reflected by menstrual cycle phase — on discordant Ki67 cases. Material and Methods: A total of 50 random cases were analyzed by 2 blinded pathologists. Ki67 expression was collected as a continuous variable. Tumors were then classified as high — or low-proliferative according to a Ki67 value of &gt; or = 15% or &lt; 15%, respectively. Sperman Rank test was used to study the correlation between initial and surgical biopsies. Data on menstrual cycle phase at the time of biopsy and surgery were retrived from medical history of all premenopausal patients.
Results: Baseline patients characteristics are as follows: median age 48 (35-56 years), premenopausal 27 (54%), postmenopausal 23 (46%), invasive ductal carcinoma 38 (76%). A differential expression of Ki67 was found in 36 patients (72%), 20 pre- and 16 post-menopausal. Spearman's Rank Order correlation did not show any significant difference between biopsy and matched surgical specimen in terms of Ki67 expression. Indeed, there was a strong positive correlation between initial and surgical biopsy, which was statistically significant across all patients (rs = .556, P &lt; 0.001), and regardless the menopausal status. However, 15% of pre-menopausal patients transformed from Ki67 low to high. Therefore, data on pre-menopausal patients were analyzed according to the phase of menstrual cycle at the timing of initial and surgical biopsies. The Spearman's Rank Order gave no correlation among 14 premenopausal patients with differential menstrual cycle phase at the timing of initial biopsy and surgery (rs = .411, P = 0.145).
Conclusions: Initial breast cancer biopsy can be used with high confidence for Ki67 determination. However, it should be noted that menstrual cycle phase could affect Ki67 expression as women with distinct menstrual cycle phase at the time of initial biopsy and final surgery do not show concordant results. Prospective evaluation of Ki67 expression in premenopausal patients is being planned.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-11-12.
P5-13-13: The Role of Topoisomerase IIa in Predicting Sensitivity to Anthracyclines in Breast Cancer Patients: A Meta-Analysis of Published Literatures.
Authors: Lu J , Du Y , Zhou Q , Yin W , Zhou L , Di G , Shen Z , Shao Z 
Published: June 2012

Abstract
Topoisomerase IIα is not only a proliferation marker of tumor cells but also a target for anthracycline-based chemotherapy. Both in vitro and in vivo studies have shown that there was a relationship between topo IIα and chemosensitivity to anthracyclines, but the predictive role of topo IIα is still controversial in breast cancer patients. A meta-analysis based on published studies was performed with the aim of obtaining an accurate evaluation of the association between topo IIα and sensitivity to anthracycline-based chemotherapy. A total of 13 eligible studies including 2,633 cases and 2,118 controls were identified. Topo IIα was associated with sensitivity to anthracyclines in locally advanced breast cancer patients who received neoadjuvant chemotherapy (RR = 1.93, 95%CI: 1.27−2.94, P=0.002; RR =1.98, 95%CI: 1.37−2.86, P&lt;0.001). In early breast cancer patients who received anthracycline-based adjuvant chemotherapy compared with non-taxane-based polychemotherapy, amplification(HR = 0.64, 95%CI: 0.49−0.83, P=0.001; HR = 0.59, 95% CI: 0.35−1.01, P=0.056) or deletion (HR = 0.82, 95%CI: 0.67−1.00, P=0.051; HR =0.58, 95%CI: 0.35−0.97, P=0.036) of topo IIα was significantly associated with better RFS and OS. The subgroup analysis in the early breast cancer patients indicated that taxane could be an interference for evaluation of the predictive role of topo IIα. In summary, the present meta-analysis suggests that topo IIα is a predictive factor for breast cancer patients who received anthracycline-based chemotherapy. Larger and well-designed prospective studies are required to further evaluate the predictive role of topo IIα in clinical practice.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-13-13.
P5-13-15: Proteomic Identification of Predictive Biomarkers of Resistance to Neoadjuvant Chemotherapy in Luminal Breast Cancer: A Possible Role for 14–3-3 and BID?
Authors: Hodgkinson VC , ELFadl D , Agarwal V , Garimella V , Drew PJ , Lind MJ , Cawkwell L 
Published: June 2012

Abstract
Background: Chemotherapy resistance is a major obstacle in effective neoadjuvant treatment for oestrogen receptor (ER)-positive breast cancer. The ability to predict tumour response would allow chemotherapy administration to be directed towards only those patients who would benefit, thus maximising treatment efficiency. We aimed to identify protein biomarkers associated with chemotherapy resistance, using proteomic analysis of fresh ER-positive breast cancer samples, and then to perform pilot clinical validation experiments. Materials and Methods: Chemotherapy resistant and chemotherapy sensitive tumour samples were collected from breast cancer patients who received standard anthracycline-based neoadjuvant therapy consisting of epirubicin with cyclophosphamide followed by docetaxel. Comparative proteomics experiments were performed using invasive ductal carcinomas which demonstrated ER-positivity (luminal subtype). Protein expression was compared between chemotherapy resistant and chemotherapy sensitive tumour samples using 2-dimensional gel electrophoresis (2-DE) with MALDI-TOF/TOF mass spectrometry (MS). In addition the Panorama XPRESS Profiler725 antibody microarray, containing 725 antibodies from a wide variety of cell signalling and apoptosis pathways, was employed in the discovery phase. Differentially expressed proteins (DEPs) were submitted to Ingenuity Pathway Analysis (IPA) to identify any canonical pathway links. A pilot series of archival breast cancer samples, from patients treated with neoadjuvant anthracycline-based chemotherapy, was used for preliminary clinical validation of putative predictive biomarkers.
Results: Five datasets were generated by antibody microarray analysis, revealing 41 targets. Of these, 7 DEPs were identified in at least 2 datasets and these included 14–3-3, BID and Bcl-xL. The top canonical pathway matched in IPA was “ERK5 signaling”, which involved 6 DEPs, including 14–3-3. The “PI3/AKT” pathway also involved 6 DEPs, including 14–3-3 and Bcl-xL. Three datasets were generated using 2-DE with MALDI-TOF/TOF MS, containing over 300 DEPs. These included several isoforms of 14–3-3. Differential expression of 14–3-3, BID and Bcl-xL was confirmed by immunoblotting in samples used for the discovery phase. A pilot clinical validation using immunohistochemical analysis of archival breast cancers revealed 14–3-3 tau and tBID to be significantly associated with chemotherapy resistance.
Discussion: We have successfully utilised clinical tumour samples for the discovery of putative biomarkers of chemotherapy resistance using two complementary proteomic platforms. We propose a potential role for 14–3-3 tau and BID as predictive biomarkers of chemotherapy resistance in ER-positive tumours and further validation in a larger sample series is now required.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-13-15.