bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-13-07: Standardized Immunocytological and Molecular Analysis of Tumor Cells and Mesenchymal Stromal Cells in Bone Marrow of Patients with Primary Breast Cancer
Authors: Sonnauer M , Kaul S , Fersis N
Published: April 2011

Background: Using criteria for classification of disseminated tumor cells in compliance with the publication of Fehm et al. (Cancer, Vol.107, 2006) we have reported a cytological DTC-positivity rate of 1.9% (39/2050) in patients with primary breast cancer (T1-4, N0/N+, M0/M1). However automated picture analysis revealed cytokeratin positive stroma cell types in DTC negative samples. Here we describe the cytological and molecular characterization of these mesenchymal stromal cell (MSC) types. Material and Methods: Mononuclear cells were enriched by Ficoll density-gradient centrifugation. Cytospin slides (mean 6) with 1x106 BM-cells were stained by CK antibodies 5D3 and A45-B/B3 using automated immunostainers with the APAAP-and the enhanced SA-FastRed-method. Slides were analysed by automated picture analysis with the ACIS I and ACIS II systems (ChromaVision). Plastic adherent cells (1x106) from culture passage one from 71 breast cancer patients were analysed for expression of CK19, MUC1 and EpCAM by FACS analysis. Trypsinized cells (n=100) were lysed and used for mRNA isolation and c-DNA synthesis (Qiagen®). A real-time quantitative RT-PCR approach (MESA FAST SYBR Assay, Eurogentec®) and primers selected from the UniversalProbeLibrary system (Roche AG®) was used for the determination of the markers EpCAM, survivin, CD144, CD146, CD 105, and CD271.

Results: Small cells characterized by an excentrically located small and clear nucleus were detected in 56% (486/866) of the bone marrow samples in a mean frequency of 1-2 cells per slide. However a subgroup of patients is characterized by elevated MSC levels. More than 5 MSC cells per 106 BM cells were detected in 19,7% (96/486) samples of the DTC negative group with elevated MSC levels in 5,2% NO, 6% N+ and 9% T3/T4 tumors. By molecular expression analysis of 71 bone marrow samples 8 probes (11%) were CK19 positive. Corresponding MSC cultures had significantly elevated expression levels of the markers vimentin, survivin, CD144, CD105, CD146 and CD271.

Discussion: We have identified CK positive bone marrow stroma cells in 56% of patients with primary breast cancer. Immunobead RT-PCR based on 107 BM cells can be used to eliminate this background and to increase DTC-sensitivity from 2 to 11%. Frequency and expression profile of MSC in these probes may give valuable additional information to understand in the complex interrelation of tumor cells with tumor-associated mesenchymal stromal cells, erythroblasts and endothelial cells in the process of metastasis.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-13-07.
Abstract P1-14-05: Predicting Brain Metastasis in Breast Cancer Patients: Who Is at Highest Risk?
Authors: Siripurapu V , Ruth K , Cristofanilli M , Egleston BL , Sigurdson ER , Freedman GM , Goldstein LJ , Bleicher RJ.
Published: April 2011

Background: The development of brain metastases (BrM) in women with breast cancer is associated with more advanced stage at presentation (Atahan 2008), although predicting who is at greatest risk remains difficult. This study aims to determine the characteristics of those at highest risk for BrM, and characterize treatment prior to their development. Methods: A prospective database containing 4,169 breast cancer patients (pts) was retrospectively reviewed for those developing BrM between 1997 and 2007. Demographic and prognostic variables were reviewed. Controls without BrM were matched for T stage, N stage and ER status, with 2 matched controls for 44 BrM pts.

Results: Forty-nine patients (1.2%) developed BrM, among whom 6 were evident at initial presentation. BrM pts overall were diagnosed at a median age of 57 y with pathologic stages I and II accounting for 33% of cases at initial presentation. Pts developed BrM at a median of 19.7 mos after diagnosis. T1/T2 tumors were present in 65% of overall cases. 21 (43%) were ER/PR negative. Among 24 sites of distant disease evident prior to BrM were 16 (66%) bone and 6 (25%) lung metastases. Univariate BrM predictors included evidence of prior non-brain metastases (p=0.0004), high nuclear grade (p=0.002), PR-negativity (p=0.005), and inflammatory breast cancer (IBC, p=0.04). Family history of breast cancer approached significance (p=0.053). On multivariable analysis for predictors, prior non-brain metastases (p=0.001), high nuclear grade (p=0.006), and IBC (p=0.01) remained significant predictors. Despite adjustment for T and N stage, univariate treatments associated with, but given prior to BrM development, were use of radiotherapy (p=0.002), chemotherapy (p=0.009), and mastectomy (vs lumpectomy, p=0.03; vs no operation, p=0.0016). The sole treatment associated on multivariable analysis was chemotherapy use (p=0.02). After BrM diagnosis, 94% had whole brain radiotherapy, 22% had chemotherapy, and 8% underwent neurosurgery. BrM pts’ median survival was 38.6 mos, while matched controls did not reach median survival in 100 mos of mean follow up. Median time from BrM diagnosis to death was 10.8 mos, with a 5 y survival of 25.6%. Conclusion: The development of BrM is associated with shorter survival in patients with breast cancer. Several clinico-pathologic factors may identify patients at greatest risk of developing BrM, even in the face of more aggressive primary therapy. Such risk factors should be considered when developing early diagnostic and therapeutic interventions for BrM, and when validating any such intervention prospectively.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-14-05.
Abstract P1-15-05: HER2 Positivity Correlates to Coexisting Invasion in Ductal Carcinoma In Situ of the Breast
Authors: Liao N , Zhang G , Liu Y , Li X , Yao M , Zu J , Xu F , Li L , Wu Y.
Published: April 2011

Background: Ductal carcinoma in situ (DCIS) of the breast with coexisting invasion is commonly seen and no consensus on any biomarkers capable of discriminating this subgroup is reached yet. We therefore carried out a retrospective study to investigate the receptor status and histological grade in Chinese DCIS patients aiming to identify if there is any independent predictor that could discriminate the subgroup of DCIS with coexisting invasion from pure DCIS patients.

Methods: A Consecutive Chinese DCIS patient cohort in a single institution was included for ER, PR and HER2 status and histological grade evaluation. Patients with invasion foci greater than 1cm in diameter were excluded. For HER2 (2+) cancers by IHC, fluorescence in situ hybridization was performed to examine HER2 gene amplification status. HER2 positive was defined as HER2 (3+) by IHC or HER2 gene amplified. Histological grading was performed only for DCIS component using Van Nuys classification system. Molecular subtypes were also profiled using the following criteria: ER and/or PR positive and HER2 negative as luminal A, ER or PR positive and HER2 positive as luminal B, ER and PR negative while HER2 positive as HER2-positive and ER, PR, HER2 negative as basal-like. Age, histological grade, ER, PR, and HER2 status were included in association analyses using appropriate statistical methods to test if any of these markers independently correlates to DCIS with invasion. Results: Our study included 183 patients in total. Among these patients, 140 had pure DCIS and 43 had DCIS with invasion. Luminal A subtype accounted for 49.7% of all cases, HER2-positive subtype for 27.9% and only 10.4% and 12.0% were luminal B and basal-like subtypes, respectively. Univariate analyses showed that histological grade 2, grade 3 and HER2 positive status were associated with DCIS with invasion, odds ratios 5.1(95%CI 1.3-19, P=0.017), 5.2 (95%CI 1.5-18, P=0.01) and 3.34 (95%CI 1.6 −6.8, P=0.001), respectively. Nonetheless, after adjusting for other markers, only HER2 positivity independently correlates to DCIS with invasion, odds ratio 3.8 (95%CI 1.4 to 10, P=0.008). Consistently, luminal B and HER2-positive subtypes were also correlated to DCIS with invasion and the odds ratio was 3.0 (95%CI 1.0-8.7, P=0.045) and 3.0 (95%CI 1.4-6.6, P=0.005), respectively when compared to luminal A subtype. Basal-like subtype was not correlated to DCIS with invasion, odds ratio 0.50(95%CI 0.11-2.4, P=0.384).

Conclusions: Our study suggests that HER2 positivity rather than higher histological grade is the independent predictor for coexisting invasion. ER/PR status and age do not correlate to coexisting invasion. Molecular subtype classification for treatment guidance especially HER2 targeted therapy in DCIS deserves further exploration.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-15-05.
Abstract P1-15-06: Incidence of HER2 Positive Quantified by FISH in Ductal Breast Carcinoma In Situ
Authors: Calvo-Plaza I , Suarez-Gauthier A , Ugidos L , Lopez-Rios F , Perez-Rodriguez F , Garcia-Garcia E , Conde E , Angulo B , Hidalgo M , Estevez L.
Published: April 2011

Purpose: The Her2/neu gene is amplified and/or its protein is overexpressed in 15-25% of invasive breast cancer, but in Ductal Carcinoma In Situ (DCIS) this expression is highly increased (60-80%). We review our rate of DCIS with Her2 amplified in a prospective series of women with new diagnosis of DCIS. Methods: Patients with new diagnosis of DCIS by biopsy were studied. The status of Her2 was evaluated by fluorescence in situ hybridization (Vysis path vision HER-2 DNA probe kit) and was considered amplified if the ratio HER2/CEP17 was >2.0. The status of estrogen and progesterone receptors were analyzed by inmunohistochemistry (IHQ). Data evaluating architectural pattern and histological grade were also collected. Results: Data from 53 patients with DCIS were included (two patients with combined invasive ductal carcinoma detected at time of surgery and 51 cases of pure DCIS). HER2 was amplified in 20 patients (37%), with HER2/CEP17 ratios between 2.16 to 14.00. A total of 29 patients had grade III DCIS, and of those 18 (62%) had Her2 amplified. 17 patients (32%) were estrogen and progesterone receptors negative (10 with Her2 amplified). We have not observed any correlation about architectural pattern (comedo, cribriform, micropapillary, solid/necrotic) and amplification of Her2. Conclusion: In our series of DCIS patients the Her2 amplification was less commonly found compared to prior studies (37% vs >60%). Most of DCIS were grade 3 (90%) and no relation was found among the architectural pattern and amplification of Her2.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-15-06.
Abstract P1-16-02: Retrospective Review of Male Breast Cancer Patients: Analysis of Tamoxifen-Related Side Effects
Authors: Pemmaraju N , Munsell M , Hortobagyi GN , Giordano S.
Published: April 2011

Background: Male breast cancer (MBC) is an uncommon malignancy with approximately 2000 American men diagnosed per year. It has been observed that the side effect profiles and tolerability with anti-hormonal treatments such as tamoxifen differ between women and men. There is a paucity of data specifically evaluating the side effect profiles of antihormonal treatments in MBC patients (pts).

Objective: To specifically study the side effects of tamoxifen in a cohort of MBC pts and to determine toxicities in this population. Methods: We retrospectively reviewed all MBC pts evaluated at our institution from 1999-2009. Out of a total of 126 MBC pts, 64 met the following inclusion criteria: stage I-III, treated with tamoxifen, and at least one follow-up visit after starting tamoxifen. A descriptive analysis of side effects was performed on these 64 pts.

Patient Characteristics and Results: Median follow-up time at our institution was 4.2 years (range 0.3-10.9). Median follow-up from the start of tamoxifen therapy was 4.0 years (range 0.3-19.4). The median age at diagnosis was 61 years (range 30-79). Breakdown by stage included: 29.7% Stage I, 54.7% stage II, and 15.6% stage III. Median initial tumor size was 2.0 cm (range 0.5-6.5). 100% were hormone-receptor positive and 7 tumors (11%) were HER2/neu-positive. Two pts had history of prior therapeutic radiation, and 11 pts (17%) had a prior or concomitant cancer diagnosis, including 6 pts with history of prostate cancer and 2 pts with history of renal cell carcinoma. Sixteen (25%) developed metastatic disease or recurrence. Thirty (47%) pts experienced at least one or more side effect while taking tamoxifen. There were a total of 101 documented side effects (more than one side effect in some pts). The most common toxicity was weight gain (14 out of 64 pts, 22%) and sexual dysfunction/loss of libido (14pts, 22%), followed by hot flashes (8pts, 13%), neurocognitive deficits (6 pts, 1%), thromboembolic events (4 pts, 1%) ocular events (3pts), mood alterations (2pts), depression (2 pts), GI disturbance(2 pts), insomnia (1) and leg cramps (1). Importantly, 13 out of the 64 (20.3%) pts discontinued tamoxifen due to toxicity: 1 for ocular, 2 for neurocognitive deficits, 4 for thromboembolic events, 2 for bone pain, 3 for loss of libido, and 1 for leg cramps.

Conclusions: To our knowledge, this is the largest retrospective study specifically examining tamoxifen-related side effects among MBC pts. Among MBC pts, there is a high rate of discontinuation of tamoxifen. Further prospective work regarding better understanding of male hormonal biology as well as novel anti-hormonal drug development may be warranted in this unique population.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-16-02.
Abstract P1-17-01: Differences in Survival among Women with Stage III Inflammatory and Non-Inflammatory Locally Advanced Breast Cancer Appear Early: A Large Population Based Study
Authors: Dawood S , Ueno NT , Valero V , Woodward W , Buchholz TA , Hortobagyi GN , Gonzalez-Angulo AM , Cristofanilli M.
Published: April 2011

Background

Significant improvements in the survival of women with breast cancer have been observed and are attributed to a multidisciplinary approach and the introduction of polychemotherapy regimens. The objective of this population based study was to determine if differences, if any, in survival exist among women with inflammatory (IBC) and non-IBC locally advanced breast cancer (LABC) diagnosed in this era.

Methods

The Surveillance, Epidemiology and End Results Registry was searched to identify women with stage IIIB and C breast cancer who were diagnosed between 2004 and 2007 who had undergone surgery and radiation therapy. Patients were categorized as either having IBC or non-IBC LABC according the AJCC 6th edition criteria. Breast cancer specific survival (BCS) was estimated using the Kaplan-Meier product limit method and compared across groups using the log rank statistic. Cox models were then fitted to compare the association between breast cancer type and BCS after adjusting for patient and tumor characteristics.

Results

Eight hundred and twenty eight (19.2%) women and 3476 (80.8%) women had stage IIIB/C IBC and non-IBC LABC respectively. Median follow up was 19 months. 2-year BCS was 90% (95% CI 88-91%) for the whole cohort and 84% (95% CI 80%-87%) and 91% (95% CI 90%-91%) among women with IBC and non-IBC LABC respectively. In the multivariable model compared to women with non-IBC LABC women with IBC had a 43% increased risk of death from breast cancer that was statistically significant (HR=1.43, 95% CI 1.10-1.86, p=0.008). Conclusion

In the era of multidisciplinary management and anthracycline and taxane based polychemotherapy regimens women with IBC continue to have inferior survival outcomes compared to women with non-IBC LABC with differences in survival appearing early.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-17-01.
Abstract P1-17-03: Comparison of Long Term Outcome of Locally Advanced and Inflammatory Breast Cancers Treated with Dose Dense Neoadjuvant Chemotherapy
Authors: Giacchetti S , Porcher R , Lehmann-Che J , Roquancourt A , Cuvier C , Hamy AS , Bertheau P , de Thé H , Marty M , Espié M.
Published: April 2011

Background: Inflammatory breast cancers (IBC) have a worse prognosis than locally advanced breast cancer (LABC) (KW Hance, JNCI 2005; 97:966-75). We report a series of LABC and IBC treated with dose dense anthracycline and cyclophosphamide and report the correlation between phenotypic features, pathological complete response (pCR) and disease free survival (DFS).

Materials and methods: Between 1990 and 2003, 196 patients (pts) with LABC (105 pts) and IBC (91 pts) treated at St Louis hospital received

6 cycles (c) of dose dense Cyclophophamid (1.2g/m2 d1)-Epirubicin (E) (75mg/m2d1) q2w (SIM regimen). 133 patients had frozen biopsy allowing p53 determination. Surgery was done after chemotherapy. pCR was defined as no residual invasive tumor in breast and lymph nodes.

Results: Median follow-up was 110 months. IBC were more often ER negative than LABC(52 % vs 34%), more often triple negative (33 % vs 22 %) and more often TP53 mutations .(60% vs 39 %).

pCR in breast and lymph nodes was identical in LABC (17 pts,16%) and in IBC (14 pts, 16 %).

Table I Pathological response and factors associated with pCR

Negative ER and presence of TP53 mutations were strong predictive factors of chemotherapy efficacy in LABC and less in IBCS. histologically grade 3 was not predictive in IBC.

Median disease free survival (DFS) was higher in LABC [103 months (m)] than in IBC (54 m), p=.031. Median Overall survival (OS) was not reached in LABC and was of 134 m in IBC (p = .015). A significant effect of ER status on DFS was found in LABC and IBC. When adjusting for ER, IBC still had a significantly shorter DFS (HR 1.64, 95%CI 1.12 to 2.39, P=.011). Similar results were found for OS. For TP 53, a significant interaction was found with LABC/IBC (p=0.017) with a risk for mutated patients in LABC (p=0.004) and a slight and nonsignificant increased risk in IBC patients (p=0.62).

Estimated disease free survival at 15 years

Conclusion: High risk patients treated with dose dense cyclophosphamide and anthracyclin have very long survival rate. Negativity of ER and p53 mutations are strong predictive factors of good outcome in LABC but less in IBC. This approach in ER-LABC should be prospectively studied..

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-17-03.
Abstract P2-02-06: Efficacy and Cost Benefit of Preoperative MRI in the Diagnostic Procedure for Operable Breast Cancer
Authors: Kojima Y , Yamauchi H , Tsunoda H , Kikuchi M , Honda S , Yoshida A , Yagata H , Tsugawa K , Nakamura S.
Published: April 2011

Purpose

The role of preoperative MRI for operable breast cancer is controversial. In this study, we evaluate the efficacy of preoperative MRI as a diagnostic tool to determine the most appropriate procedure (i.e; mastectomy or partial mastectomy) for operable breast cancer patients and cost saving benefit from selecting the most appropriate procedure.

Patients and Methods

From January 2006 to December 2007, we retrospectively studied 1149 patients. All of these patients were diagnosed with breast cancer and consequently underwent mammography (MMG), ultrasound (US) and MRI before surgery so as to determine the optimal procedure. We extracted all the cases which required changes in procedures due to the MRI findings, and compared the radiological data to the clinical and pathological outcomes. Furthermore, we studied the overall cost differences between using MRI and not using MRI. Results

In seventy seven cases, 6.7% of the total, operational changes were made because of preoperative MRI findings, which resulted in better operational outcomes. The main reason for procedural changes was the MRI findings revealed more extensive lesions, which pathologically proven to be in situ lesions. In some cases, additional lesions were pointed out by the MRI, which led us to avoid undertreatment. Among 77 cases, 69 were pathologically proven to have accurate radiological diagnosis and appropriate procedural change with MRI. The positive predictive value of preoperative MRI for partial resection was 99.1% and that for total mastectomy was 97.1%. The calculated cost difference for appropriate procedural change with MRI compared with not using MRI was $4,340. From our data, in 69 cases among 1149 received this appropriate change, $260 per case would be saved, which assumed to be more than $130 million in a year.

Conclusion

Adding preoperative MRI in the diagnostic procedures for the accurate operation in breast cancer might be effective. Usage of MRI preoperatively also save the inappropriate cost in breast cancer operations.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-02-06.
Abstract P2-02-08: 3-Tesla MRI Is More Useful for the Evaluation of Intraductal Spreading Area of Breast Cancer: A Comparative Study with Conventional 1.5-Tesla MRI
Authors: Tanaka H , Kubo M , Kamitani T , Yamamoto H , Shirahane K , Tanaka M.
Published: April 2011

Background: High quality of space and temporal resolution is demanded from the diagnosis for intraductal spreading area of breast cancer. In our hospital, from April 2007, 3-Tesla (T) MRI was transduced and high resolution dynamic studies for bilateral mammary have been performed. We have reported the accuracy of 3-T MRI in the evaluation of intraductal spreading area of breast cancer at 31st SABCS. At this time, we demonstrate our follow-up data.

Methods: We assessed 256 patients with primary breast cancer who underwent the 3-T or 1.5-T breast MRI as pre-operative examination between April, 2007 and February, 2010. Concerning the diagnosis of intraductal spreading area of breast cancer, we weighed 3-T MRI against conventional 1.5-T MRI. Using 1.5-T or 3-T MRI (Achieva, Philips), we injected contrast medium (Magnepist) with 0.2ml/kg body weight in 1ml/sec and photographed five aspects before injection and after injection for every 90 seconds.

We obtained fat-sat T1-weighted magnetic resonance image (T1WI), multi-planar reconstruction (MPR) image and standard maximum intensity projection (MIP) image based on T1WI by coronal section, and furthermore slab MIP image with Aquerius net (Terarecon), and we evaluated the enhanced area.

Results: 138 cases were examed with 1.5-T MRI, and 118 cases with 3-T MRI. In 1.5-T group, 40 cases (30%) were underestimated, additional resection were required intraoperativelly with cancer-cells positive surgical margin in 18 cases (13%), and surgical margin was positive with carcinoma cells by final pathological findings in 12 cases.

In two patients we were forced to changes from breast-conserving surgery to mastectomy, and two cases underwent reoperation (mastectomy) with the diagnosis of widely positive surgical margin. In comparison with 1.5-T MRI group, 16 cases (14%) were underestimated in 3-T MRI group, intraoperative addition resection in eight cases (7%), and surgical margin positive by the final pathological findings in 13 cases, respectively.

Conclusions: The concern to enhance an artifact is known on 3-T MRI because it has high resolution. However, it may be more useful rather than1.5-T MRI because of its high resolution, to evaluate the intraductal spreading area of breast cancer and to determine the extent of surgical resection.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-02-08.
Abstract P2-02-10: Clinical Utility of MRI before Re-Lumpectomy in Breast Conserving Operation
Authors: Jeon C , Lee JS , Kim S
Published: April 2011

Background: It was necessary to know the resection margin status for breast conserving operations after diagnostic excisional biopsies. The purposes of this study were to assess the accuracy of breast MRI in detecting residual tumor extent around excisional biopsy or mammotome biopsy sites with unknown resection margin status and evaluate the impact of MRI on surgical treatment planning for breast conserving operation. Material and Methods: Preoperative breast MRI was performed in 47 breast cancer patients who underwent mammotome biopsy and excisional biopsy with unknown resection margin status. All subjects underwent breast MRI and ultrasonography for extent of residual tumor, and were evaluated by experienced two diagnostic radiologists, followed by immediate curative operation. We investigated the correlation between radiologic and pathologic finding and the radiologic decision's effect on further surgical treatment. Results: 28 patients(59.6%) were taken by excisional biopsy, 13 patients by mammotome biopsy(27.7%), and 6 patients(12.8%) by lumpectomy. Twenty-five patients (53.1%) underwent re-lumpectomy, while 19 patients (40.4%) mastectomy, and 3 patients(6.3%) only sentinel lymph node biopsy. Residual tumor was present in 33 patients (70.3%) and multifocal breast cancer was found in 12 patients (25.5%). In terms of the accuracy of MRI in detecting residual tumor, the sensitivity was 63.6%, specificity was 45.5% and the positive predictive value was 77.7%. In that of ultrasonographies, the sensitivity was 52.3%, specificity was 88.8%, and the positive predictive value was 91.6%. Pathologic finding of residual tumor showed that ductal carcinoma in situ was most common (57.1%) in not only radiologic-pathologic concordant cases, but also in radioloigc-pathologic discordant cases (58.3%). But 5 patients (10.6%) with invasive ductal carcinoma in residual tumor were not detected by MRI. Positive predictive value was not associated with residual tumor sizes. The MR findings of residual tumor changed the next procedure from re-lumpectomy to mastectomy (10 patients), additional biopsy of suspicious lesion in ipsilateral breast (2 patients), and second-look ultrasonography (4 patients). Discussion: Preoperative MRI was not critical effect on evaluating the margin status around excisional biopsy sites. An important clinical role of

MRI is to identify a subset of patients with extensive residual or multifocal breast cancer who would be better served by undergoing mastectomy rather than breast conserving operation.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-02-10.
Abstract P2-04-01: HER2 Status as Predictor of Mammographic Screening Detection: Comparison of Interval-and Screen-Detected Breast Cancers
Authors: Musolino A , Michiara M , Conti GM , Boggiani D , Bozzani F , Zatelli M , Sgargi P , Ardizzoni A.
Published: April 2011

Background: To determine whether markers of poor prognosis are associated with risk of breast cancer diagnosis in the interval between screening examinations, we estimated the effect of the mode of detection on distribution of breast cancer molecular subtypes using population cancer registry data.

Material and Methods: Subjects (n = 641) comprised all breast cancers systematically collected by the Cancer Registry of Parma Province and diagnosed in women aged 50-69, from 2004 to 2007. These included 370 screen-detected and 271 symptomatic breast cancers (63 women with interval cancers and 208 who had not attended screening). We used logistic regression to determine whether interval cancers were associated with selected clinical and biologic characteristics. We also estimated the relative risk of cause-specific fatality and disease-free survival (DFS) by each resulting predictive factor (screen-detected compared to either symptomatic or interval cancers).

Results: Interval-detected cancers occurred more in younger women and were of more advanced tumor stage than screen-detected cancers. In unconditional logistic regression models adjusted for age and tumor stage, tumors with high histologic grade (odds ratio[OR] = 2.2; 95% CI =1.0-5.4), high proliferation rate (OR =2.7; 95% CI =1.5-4.8), or positive HER2 status (OR =2.6; 95% CI =1.3-5.1) were more likely to surface in the interval between screening examinations. After adjusting for various potential biases, women with screen-detected breast cancer had a substantial survival advantage over those with symptomatic breast cancer. In a multivariate model, positive HER2 status independently predicted poor DFS when the mode of cancer detection was included as covariate in addition to age, histologic grade, proliferation rate, and tumor stage. Conclusions: This is the first population-based cancer registry study demonstrating that HER2-positive tumors account for a substantial proportion of mammographic screening failure to detect breast cancer. Our data indicate that molecular subtype distribution of screen-detected breast cancer differs from that of interval cancers and accounts in part for the better outcome of screen-detected cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-04-01.
Abstract P2-04-02: Increased Risk of Breast Cancer in Women Who Receive False Positive Test Results from Mammography Screening
Authors: von Euler-Chelpin M , Larson Thorsted B , Vejborg I
Published: April 2011

Background: Copenhagen has the longest running organised screening programme for breast cancer in Denmark. The programme started in 1991. One of the possible consequences when screening for breast cancer with mammography is a false positive test result. We assessed the relative risk of developing breast cancer among women who had received false positive test results compared to women who had not.

Method: The mammography programme provided data on participation and test results from 1991 -2005. Oncological data was retrieved from the National Cancer Register. Follow-up ended on 17 April 2008. Data were linked using the Danish unique personal identity number. Relative risks were estimated using Poisson regression, age adjusted. The exposure was categorised as a) false-positive Type1 = negative after further assessment with mammography, ultrasound and fine needle aspiration, and b) Type 2 = negative after surgery.

Results: A total of 59.392 women participated in the mammography programme, representing 650.218 person-years at risk. The relative risk (RR) of breast cancer after any type of false positive (FP) was 1.33 (95% CI, 1.18-1.50), the RR for breast cancer after a Type 1 FP was 1.34 (95% CI, 1.18 — 1.52) and after a Type 2 FP the RR was 1.24 (95% CI, 0.85-1.82). The RR of interval cancer, i.e. breast cancer within the two years following the test result, was 1.73 (95% CI, 1.13-2.66) for women who had received a FP test result as compared to those who had not. Conclusion: Women who have received a false positive test result from mammography screening had a 73% increased risk of developing interval cancer compared to women who had not, and a 33% increased risk of developing breast cancer at any time after a false positive test result. There was an increased risk for both Type 1 (34%) and Type 2 (24%) FP test results, though data was only significant for Type 1 results.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-04-02.
Abstract P2-04-03: Usefulness of Perioperative Screening of Patients with pN0 Invasive Breast Carcinoma: Results on 1058 pN0 Patients of a Population-Based Study of the French Cancer Registry
Authors: Frenel J-S , Leux C , Molinie F , Campone M
Published: April 2011

Background: Usefulness of baseline routine bone scanning (BS), liver ultrasonography (LUS) and chest radiography (CXR) in asymptomatic women with pN0 Invasive Breast Carcinoma is not evidence-based. Methods: A random sample of 1883 cases of invasive breast cancer diagnosed in 2003 from 10 French counties covered by a cancer registry was analyzed. Clinico-pathological features, staging tests and results were retrospectively collected from the medical record. Results: 1058 pN0 patients were identified. At baseline, CXR, LUS, BS were carried out pre or post-operative in respectively 785 (74%), 782 (74%), and 618 (58%) patients. Data summarizing results with the three imaging techniques are listed in [Table 1]. Initial suspicion was confirmed by other independant tests (bone X-ray, computerized tomography scan). After the initial staging investigation procedures and the further diagnostic procedures if necessary (computerized tomograhy scan or bone X-ray), only 3 (0.3%) patients were diagnosed having metastatic disease. CXR, LUS, BS detected metastasis respectively in 1 (0.13%), 0 (0%), and 2 (0.3%) of patients.

Diagnostic accuracy in the diagnosis of metastatic disease by chest radiography, liver ultrasonography and chest radiography

Conclusion: In asymptomatic pN0 invasive breast cancer, screening for metastases is not warranted because of low frequency of metastases, false positive findings and high cost.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-04-03.
Abstract P2-06-14: Utilizing Human Breast Tissue and Blood to Discern Novel Protective Properties of Early Pregnancy in Reducing Breast Cancer Risk
Authors: Gutierrez CM , Mills DJ , Love S , Lakshmanaswamy R.
Published: April 2011

Introduction: The incidence of breast cancer, the most common cancer in women, is very high in Western Countries and is increasing rapidly in the Third World. Yet, despite better screening, earlier diagnosis, and the development of new therapies, a cure for metastatic breast cancers remains an elusive goal. Since it is difficult to defeat breast cancer after it has occurred, finding new strategies to delay or prevent the development of breast cancer has been an important area for clinical and experimental investigations. Multinational studies have suggested that women who undergo a full-term pregnancy before the age of 20, with or without lactation, have about one half the risk of developing breast cancer compared to women who undergo their first pregnancy after the age of 35 or women who have never undergone a full term pregnancy. Thus, pregnancy at an early age protects against breast cancer development. This universal protective effect of pregnancy serves as a major clue and starting point in the search for new experimental strategies and novel biomarkers related to the prevention of breast cancer.

Methods: Women were recruited who had never been diagnosed with breast cancer and fit in one of the following categories; 1) had their first child ≥25 years of age, 2) had their first child ≥35 years of age, and 3) a control group of women the same age as those in groups 1 and 2 but who have never given birth were recruited and provided a sample of breast tissue and blood simultaneously. These groups were categorized as early parous, late parous, or nulliparous and their RNA was isolated from tissue and whole blood. Human whole genome microarray analysis (Agilent) was performed to identify the genes that were persistently altered in expression due to early pregnancy.

Results: The study of molecular alterations in breast cancer genes has succeeded in identifying some genetic mechanism of breast cancer development. We attempted to elucidate early pregnancy-related changes in genes in breast tissue and blood associated with resistance to breast cancer. Over 2200 genes were up-regulated 2 folds or more in the blood and breast of early parous women than late parous and nulliparous women. Approximately 5000 genes were down-regulated 2 folds or more in the blood and breast of early parous women compared to late parous and nulliparous women.

Conclusion: These studies focused on the analysis of human breast tissue and blood to gain a deeper understanding of changes in parity associated with the protective effect of early pregnancy in women. From these altered gene expressions we have identified a gene signature for early parity. The persistently altered genes will be used as novel biomarkers to predict breast cancer risk and develop strategies to protect the breast from cancer development.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-14.
Abstract P2-06-20: Hormonal Criteria for Assessment of Menopause: Role of Anti-Mullerian Hormone
Authors: Chatterton RT , Khan SA , Heinz RE , Kmiecik T , Scholtens D.
Published: April 2011

Introduction: The difficulty in establishing the time of menopause poses a problem in many breast cancer research studies. We sought to determine the value of adding anti-mullerian hormone (AMH) to the accuracy of designating hormonal signs of menopause in 179 women who were recruited for a case-control study of nipple aspirate fluid (NAF) biomarkers.

Methods: A questionnaire was administered to obtain medical and reproductive history, self-reported menstrual cycle length, last menstrual period and age. Serum estradiol, progesterone, FSH, and AMH were measured in a blood sample obtained on the day of the study encounter. Criteria for premenopause were: serum FSH <30 mIU/ml, AMH >0.08 ng/ml, estradiol (E2) >30 pg/ml, and menstrual periods of 25 to 35 days. For postmenopause the inverse of the hormone levels and no menstrual periods for >12 mo and age >52 years was used. Serum progesterone (P4) concentration of <3 ng/ml provided confirmation of postmenopause. Results: The age range was from 35 to 69 years (median 51). By self-report (amenorrhea >12 mo and age) 76 subjects were premenopausal; 70 were postmenopausal and 33 subjects were perimenopausal. Table: Designation of menopausal state based on hormonal criteria: Concordance of individual measures with the designation based on all criteria (percentages).

Table

FSH was found to be discordant in 1 (1.1%) of premenopausal subjects and in 5 (5.3%) of postmenopausal subjects. AMH was found to be discordant in 21 (24.7%) of premenopausal subjects and 6 (6.4%) of postmenopausal subjects. Estradiol was discordant in 13 (15.3%) of premenopausal subjects and in 7 (7.4%) of postmenopausal subjects. Progesterone was <3 ng/ml in all of the postmenopausal subjects. The combination of FSH and AMH predicted 100% of the grouping based on all measures; FSH and E2 predicted 99.9%. The absence of progesterone is a certain criterion for postmenopause but adds nothing to the designation of premenopause. Comment: An accurate determination of the hormonal and reproductive status of women is critically important in studies relating hormones to breast cancer risk. The loss of ovarian hormones has long-term implications, and the menopausal status at the time of sampling determines the recent and current hormonal exposure. Using the hormonal criteria, all women in this study could be classified as pre-or postmenopausal. Adding AMH to the usual hormones did not improve the designation based on the other criteria.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-20.
Abstract P2-06-22: The Modulation of CXCL9 Release by Prostaglandin E2 and Cyclooxygenase Inhibitors in Human Breast Cancer
Authors: Bronger H , Kräft S , Stöckel A , Welk A , Kiechle M , Schmitt M , Schwarz-Boeger U.
Published: April 2011

Introduction

IFN γ inducible chemokine CXCL9 impairs tumor growth and metastasis formation through recruitment of natural killer (NK) cells and tumor suppressive T lymphocytes in murine breast cancer models. In human breast cancer CXCL9 mRNA expression correlates with response to different chemotherapeutic regimens and tumor infiltrating lymphocytes, indicating that enhancing intratumoral CXCL9 is a feasible therapeutic way to improve tumor suppressive immune infiltration. However, little is known about the regulation of CXCL9 in human breast cancer. Since overexpression of cyclooxygenases (COX) correlates with impaired NK cell infiltration as shown before, this system is a conceivable candidate for modulating intratumoral CXCL9 concentration. Material and Methods

For regulation studies, MCF 7 and MDA MB 231 breast cancer cells were stimulated with IFN γ with or without prostaglandin E2 (PGE2) or COX inhibitors (indomethacin, aspirin, celecoxib). CXCL9 was localized in human breast cancer samples by immunohistochemistry. Its release was measured by ELISA. Expression of CXCL9 and cyclooxygenases in 60 breast cancer samples was determined by ELISA and western blot, respectively.

Results

Within the tumor microenvironment, the cancer cells are the major source of CXCL9, thus making them the appropriate cell type to study CXCL9 regulation. PGE2 significantly reduces IFN γ mediated CXCL9 release from MCF 7 and MDA MB 231 cells, and inhibition of endogenous cyclooxygenases by indomethacin or aspirin correspondingly augments this secretion. However, the COX 2 specific antagonist celecoxib has PGE2-agonistic effects and impairs CXCL9 release, probably via COX independent mechanisms.

Conclusion

Suppressing endogenous PGE2 by cyclooxygenase inhibition increases CXCL9 release from breast cancer cells and is therefore a feasible way to enhance intratumoral immune infiltration. However, due to possible COX independent mechanisms the choice of the appropriate COX inhibitor seems to be crucial. The predictive value of CXCL9 as a novel biomarker and its correlation with COX overexpression in human breast cancer is discussed.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-22.
Abstract P2-06-23: A New Way To Determine Multifocality: The Use of Molecular Markers?
Authors: Werkoff G , Antoine M , Rodenas A , Gligorov J , Chopier J , Uzan S , Rouzier R.
Published: April 2011

AIM: Multifocality in breast cancer has a great on treatment options. However, the prevalence of multifocality is currently underestimated at the time of diagnosis because of the lack of sensitivity of clinical examination and imaging. Our aim was to develop a tool for predicting multifocality based on clinical and immunohistochemical criteria. METHOD: This study was conducted at Tenon Hospital with samples from patients operated on between 2002 and 2009. We constructed a tissue micro-array (TMA) from 100 breast specimens: 50 multifocal tumors and 50 unifocal unifocal tumors. Unifocal tumors were matched with multifocal tumors by size of largest focus, hormonal receptor status and SBR grade. These unifocal controls had either a mastectomy or a pre-operative MRI to rule out occult multifocality. We studied the expression of ten different antibodies (HER2, KI67, CK 5/6, CK 14, CK18, Vimentine, C-kit, EGFR, p53, and p63). From the results of this analysis we developed a prediction model based on a multivariate logistic regression. RESULTS : For the proteins tested, the concordance of immunohistochemical staining of foci from multifocal tumors ranged from 72% to 100%. In the univariate analysis, only c-kit expression was significantly associated with multifocality (P<0.05). The discrimination (area under the ROC curve) of a multivariate model based on clinical, pathological, and immunohistochemical variable was good (0.72) but lacked robustness when tested by internal validation.

CONCLUSION: We report the first clinico-biological predictor of multifocality. New markers are currently tested to improve the performance of the multivariate model.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-23.
Abstract P2-07-01: Simvastatin Prevents Breast Cancer Skeletal Metastasis by Increasing p53 Levels To Increase PTEN and Inhibit CD44 Expression
Authors: Mandal CC , Ghosh-Choudhury N , Yoneda T , Choudhury GG , Ghosh-Choudhury N.
Published: April 2011

Bone represents one of the main sites of metastatic spread of breast cancer. The prognosis for the breast cancer patients with bone metastasis is very poor with severely compromised life style associated with extreme bone pain and fracture prevalence. We used heart injection model to test the effect of simvastatin on breast cancer bone metastasis. Simvastatin prevented osteolytic lesions in mice injected with MDA-MB-231 (MDA) human breast cancer cells, indicating that statin attenuates breast cancer metastasis to bone. To investigate the mechanism, MDA cells were used in an in vitro wound-healing assay. Simvastatin significantly inhibited the migration of MDA cells. Transwell invasion assay using collagen-coated discs confirmed the inhibitory effect of simvastatin on breast cancer cell invasion. We have recently shown that simvastatin inhibits primary tumor growth in mice by upregulating the tumor suppressor PTEN. Overexpression of PTEN in MDA cells significantly blocked their ability to migrate and invade. To determine the mechanism of PTEN upregulation, we considered the tumor suppressor protein p53, which is known to regulate PTEN expression. Simvastatin-treated mice tumors showed significantly increased levels of p53. Similarly, simvastatin enhanced the expression of p53 in MDA cells. Additionally, simvastatin increased transcription of the reporter plasmids containing the consensus p53-binding element (p53-Luc) or the PTEN promoter (PTEN-Luc). Cotransfection of p53 with these reporter constructs resulted in similar increase in transcription, indicating a role of p53 in PTEN expression by simvastatin. Recent reports indicate a direct correlation between expression of CD44 in breast cancer and their metastasis to other organs including bone. Levels of CD44 were significantly reduced in the tumors of mice treated with simvastatin. In MDA cells, simvastatin decreased the expression of CD44 protein and its transcription, as determined by CD44 promoter-driven luciferase construct (CD44-Luc). p53 acts as transcriptional activator or transcriptional repressor in a cell-type and context-dependent manner. Presence of non-canonical p53-binding elements has been reported in the CD44 promoter. Cotransfection of p53 with CD44-Luc significantly suppressed the transcription of CD44. Furthermore, expression of p53 markedly reduced the expression of CD44 protein. Finally, downregulation of endogenous CD44 by specific shRNA decreased migration of MDA cells. Together these results uncover a novel action of simvastatin, which targets p53 expression to simultaneously increase PTEN and decrease CD44 thus blocking migration and preventing bone metastasis of the human breast cancer cells.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-07-01.
Abstract P2-07-02: Low-LET γ-Radiation Induced Protein S-Nitrosylation Regulates Tumor Invasion and Metastatic Transcriptional Response
Authors: Veeraraghavan J , Natarajan M , Herman TS , Aravindan N.
Published: April 2011

Local-regional radiotherapy is one of the major therapeutic means in the management of breast cancer. However, besides the therapeutic effect, studies have shown that irradiation promotes malignant behavior of cancer cells both in vitro and in vivo by activating several tumor invasion and metastatic molecules. In this regard, ascertaining the molecular alterations contributing to this radiation-induced response is imperative. Accordingly, we investigated the role of radiation-induced NO-dependent nitrosylation in the transcription of invasion and metastatic genes in human breast adenocarcinoma cells. Human MCF-7 cells either mock irradiated, exposed to 2Gy (IR), treated with L-NAME (NOS inhibition) or BAY-11-7082 (phosphorylation inhibition) and then exposed to 2Gy, were examined for modulations in the transcriptional response of 93 tumor invasion and metastasis signaling genes. Cells treated with β-estradiol and mock-irradiated MDA-MB-231 cells were used as positive controls. Compared to mock irradiated cells, real time QPCR profiling revealed that 20 of 29 IR-induced genes including Birc3, Casp8, Ctgf, Ctsk, Htatip2, Kras, Map2k7, Mgat5, Mta2, Nme1, Nme2, Nme4, Pax5, Pten, Runx1, Serpinb5, Syk, Timp1, Timp2 and Timp4 were completely suppressed with NOS inhibition. Conversely, as opposed to NOS inhibition, IR-induced phosphorylation inhibition either failed to suppress or further induced the expression of these IR-induced genes. Furthermore, β-estradiol treatment revealed upregulation of 16 of these 20 IR-induced nitrosylation-dependent molecules and serves as a positive control for the study. More importantly, the highly metastatic, MDA-MB-231 cell line showed a robust induction in the expression of 15 of these 20 IR-induced nitrosylation-dependent genes. Taken together, these data imply that IR induces the expression of tumor invasion and metastatic genes and more importantly, may regulate the radiation-activated invasion and metastatic phenotype in breast adenocarcinoma cells.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-07-02.