bluuebunny/crossref_metadata_2025 huggingface dataset:
P3-02-02: Novel Ranpirnase-Based ImmunoRNases Display Potent Cytotoxicity in Diverse Human Breast Cancer Cell Lines.
Authors: Goldenberg DM , Liu D , Wang Y , Rossi EA , Chang C-H
Published: June 2012

Ranpirnase (Rap) is an amphibian ribonuclease originally isolated from the oocytes of Rana pipiens. Rap shows anti-tumor activity in diverse cancers and its potency can be enhanced by chemically linking or recombinantly fusing Rap to a tumor-targeting antibody, as demonstrated for CD22- or CD74-expresing hematological malignancies, as well as a variety of Trop-2-expressing cell lines derived from breast, cervical, lung, pancreatic, and prostate cancers. The Dock-and-Lock (DNL) platform enables the design and generation of targetable therapeutics that are multivalent, multispecific, and multifunctional. Here, we report the successful application of the DNL method to generate a novel class of Rap-based immunoRNases, each of which features a pair of dimeric Rap molecules covalently tethered to a select monoclonal antibody at the carboxyl termini of the heavy chains. Two such constructs, designated E1-Rap and 22-Rap, were developed with hRS7 (humanized anti-Trop-2) and epratuzumab (humanized anti-CD22), respectively, purified to near homogeneity, and evaluated in a panel of human breast cancer lines, including the basal-like, triple-negative subtype (MDA-MB-468, MDA-MB-231, BT20, HCC1806, and HCC1395), the luminal B, HER2−negative subtype (MCF-7), and the HER2−positive subtype (SKBR3), all except HCC1395 expressing high to moderate levels of Trop-2, and none expressing CD22. As demonstrated by flow cytometry, E1-Rap and hRS7 bound equivalently to MDA-MB-468, indicating the affinity of E1-Rap for Trop-2 is not compromised. Surprisingly, 22-Rap, but not epratuzumab, also bound substantially to MDA-MB-468, albeit to a lesser extent than E1-Rap. We thus postulate that the four highly basic Rap molecules in the DNL conjugate may confer a spatial configuration to largely enhance their interaction with negatively-charged cell surface proteins, such as heparan sulfate proteoglycans. Whereas the individual DNL component (IgG or Rap) alone or in combination showed negligible in vitro cytotoxicity in all seven breast cancer cell lines examined, E1-Rap exhibited EC50 values of 1 nM or less in MDA-MB-468 (0.03 nM), MCF-7 (0.1 nM), BT20 (0.18 nM), HCC1806 (0.19 nM), and SKBR3 (1.29 nM). In comparison, the potency of 22-Rap was at least 10-fold lower than E1-Rap in MDA-MB-468, BT20 and HCC1806, with an EC50 of ∼2 nM. Neither E1-Rap nor 22-Rap was very effective in inhibiting the proliferation of the more aggressive MDA-MB-231, with an EC50 above 50 nM. In the Trop-2-negative HCC1395, the dose-response curves obtained for E1-Rap and 22-Rap were nearly identical (EC50 ∼100 nM), as they should be if the cytotoxicity was mediated mainly through the Rap component. The results of Immunofluorescence microscopy showed E1-Rap was effectively internalized and localized in the cytosol. Thus, these ImmunoRNases are potentially new cancer therapeutics for breast and other solid tumors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-02-02.
P3-03-01: Low Expression of microRNA-210 Is an Independent Good Prognostic Factor in Japanese Triple-Negative Breast Cancer Patients.
Authors: Toyama T , Kondo N , Endo Y , Sugiura H , Yoshimoto N , Iwasa M , Takahashi S , Iwase H , Fujii Y , Yamashita H
Published: June 2012

Background: microRNAs (miRNAs) have emerged as a new class of non-coding genes involved in regulating cell proliferation, differentiation, and viability. Recent studies have identified miR-210 among a set of hypoxia-regulated miRNAs and demonstrated a direct regulatory role of hypoxia-inducible factor-1 alpha (HIF-1 a) in its transcription. High expression of miR-210 has been reported to be a poor prognostic factor in several types of cancers including breast.

Materials and Methods: TaqMan MicroRNA assays for miR-210 expression were performed in 219 breast cancers (58 triple-negative (TNBCs), and 161 ER-positive and HER2−negative). Correlations between miR-210 expression and clinicopathological factors were analyzed. The effects of several variables on survival were tested by Cox proportional hazards regression analysis.

Results: miR-210 expression in TNBCs was significantly higher than in ER-positive and HER2−negative breast cancers (p<0.001). Patients whose TNBCs had low miR-210 expression experienced significantly better disease-free and overall survival compared with high miR-210 expressors (p=0.02 and p=0.05, respectively). Notably, among 40 node-negative TNBCs, 5-year disease-free survival was approximately 60% in patients whose tumors had high or intermediate miR-210 expression (n=26), while no patients with low miR-210 expression (n=14) suffered recurrent disease. Cox univariate and multivariate analyses demonstrated that low expression of miR-210 was an independent good prognostic factor in TNBCs.

Discussion: Although prognosis of patients with TNBCs is poor, those whose tumors expressed low levels of miR-210 had a more favorable prognosis. Thus, the degree of miR-210 expression might be a clinically useful prognostic factor for decision-making regarding treatment in the adjuvant setting, especially in node-negative TNBC patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-01.
P3-03-03: Congruence between Patterns of microRNA Expression and Histologic Grading of Invasive Breast Carcinomas.
Authors: Ellsworth DL , Croft DT , Field LA , Deyarmin B , Kane J , Ellsworth RE , Hooke JA , Shriver CD
Published: June 2012

Background

Histologic grading may be used as an indicator of prognosis in breast cancer; patients with low-grade carcinomas have ∼85% ten-year survival compared to just 45% survival in patients with high-grade disease. Although useful for risk stratification, assigning nuclear grade is subjective, and a large proportion of carcinomas are classified as intermediate-grade with uncertain prognosis, thus limiting clinical utility. MicroRNAs (miRNAs) regulate gene expression and serve an important role in breast cancer development. In this study we examined miRNA expression profiles in low-grade and high-grade breast carcinomas to determine if miRNA expression is associated with pathological classifications of tumor grade.

Methods: Breast tumors were obtained from 69 patients enrolled in the Clinical Breast Care Project. Samples were partitioned into low-grade (n=30) or high-grade (n=39) categories using the Nottingham Histologic Score. Following laser microdissection of frozen tissue sections, miRNA was isolated from pure populations of breast tumor cells and hybridized to Affymetrix GeneChip® miRNA arrays containing over 800 human miRNA probes. Expression profiles were analyzed with Partek Genomics Suite using the miRNA Expression Module.

Results: We identified 30 unique miRNAs that showed differential expression at a False Discovery Rate (FDR) p<0.05 between low-grade and high-grade breast carcinomas. Gene targets for these miRNAs function primarily in metabolism and cell communication. Expression of hsa-miR-18a and hsa-miR-572 was significantly different between histologic grades at an FDR p<1×10−8 and hierarchical clustering based on these miRNAs correctly classified 97% (29/30) of low-grade and 90% (35/39) of high-grade tumors. miR-18a has been shown to inhibit ER signaling and promote cellular differentiation, while the role of miR-572 in breast carcinogenesis is not well known.

Conclusions: Dysregulation of miRNAs may accompany changes in cellular morphology typically used in histologic classification of breast carcinomas. Patterns of miRNA expression may improve reproducibility and clinical utility of tumor grading and may prove useful for prediction of recurrence and survival for patients with intermediate-grade carcinomas.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-03.
P3-03-07: A Novel Approach Integrating microRNA and mRNA Signatures of HMAPK Signaling Is Highly Predictive of ER- Status and Outcome in Breast Cancer – Role of HMAPK microRNAs in Repression of ER and p27.
Authors: Miller P , Clarke J , Koru-Sengul T , Brinkman J , El-Ashry D
Published: June 2012

Deregulation of the MAPK signaling pathway in breast cancer is known to facilitate the down-regulation of the estrogen receptor and to contribute to the aggressive nature of ER negative and triple negative breast cancers. We have identified a microRNA signature indicative of hyperactive MAPK (hMAPK) signaling, which complements a previously established hyperactive MAPK gene expression signature. We have shown that hMAPK signaling also alters the regulatory activity of many microRNAs, including particular microRNAs with established roles in the biology of breast cancer, miR-221/222 and miR-22. Expression correlation with both the hMAPK microRNA signature and hMAPK mRNA signature is significantly associated with ER-negative status, increased tumor grade, high proliferation rate, and, importantly, poor disease specific survival among breast cancer patients, regardless of ER status. The hMAPK microRNA signature contains 127 microRNAs, 47 up-regulated and 70 down-regulated. Of note, hMAPK up-regulated microRNAs include miR-221/222 and 22, both of which have been demonstrated to target ER while miR-221/222 targets the cell cycle regulatory protein p27. Down-regulated microRNAs include miR-375, which positively regulates ER expression by down-regulating expression of an ER repressor. miR-221/222 and miR-22 exhibit both enhanced expression and enhanced regulatory activity in the context of hMAPK signaling, indicating an important role for these microRNAs in the biology of hMAPK signaling in breast cancer. In addition to these microRNAs, an unbiased approach of determining MAPK regulated microRNAs targeting the 3’ UTRs of both ER and p27 will identify novel microRNAs involved in the MAPK regulated repression of ER and p27. These data not only suggest a regulatory role for microRNAs whose expression and biological activity are altered under conditions of hyperactivation of MAPK signaling in establishing and maintaining ER negativity and tumor aggression, but also indicate that hMAPK signaling may represent a novel aggressive tumor biology that is indicative of poor disease outcome in breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-07.
P3-04-04: Patterns of Distant Metastasis According to the Molecular Subtypes of Breast Cancer; Results of 529 Breast Cancer Patients.
Authors: Hwang MJ , Seo YJ , Lee JH , Son GT , Choi JE , Bae YK , Kang SH , Lee SJ
Published: June 2012

Backgrounds; Distant metastasis from breast cancer arises from various sites. But few studies concerning factors that can predict metastasis patterns in breast cancer has been reported. In this study, we analyzed the effect of breast cancer molecular subtypes on distant metastasis patterns and tried to determine factors that predict metastasis sites.

Patients and methods; From January 1995 to January 2004 at Yeungnam university hospital, patients diagnosed with the primary invasive breast cancer and received treatments were included in this study. Patients with bilateral breast cancer or distant metastasis at diagnosis were excluded. After analyzing estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor 2 (HER2), epidermal growth factor (EGFR) and cytokeratin (CK) 5/6 status, we classified patients into 5 categories, luminal A, luminal B, HER2−enriched, basal-like, and normal (triple negative nonbasal) breast cancers. Distant metastatic patterns of each category were analyzed.

Results; 529 patients were eligible for tissue microassay analysis and median follow-up period was 7.7 years. In this period, total 82 patients (15.5%) had locoregional relapse or distant metastasis and distant metastasis were identified in 54 patients (10.2%). Each distant metastatic rate was 10.5% (33/313) in luminal A, 8.7% (4/46) in luminal B, 10.2% (6/59) in HER2−enriched, 7.9% (7/89) in basal-like, and 18.2% (4/22) in normal subtype. Most frequent site of distant metastasis in all patients was bone and such result was consistent with Luminal A subtype. Liver metastasis was most frequent in Luminal B subtype, lung metastasis in HER2 enriched and normal subtype and brain metastasis in basal-like subtype.

Conclusions; There was a definite association between breast cancer molecular subtype and distant metastatic pattern. If more patients and prolonged follow up periods are analyzed, we would be able to determine the best follow up intervals, methods and treatment directions concerning subtypes of breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-04.
P3-04-06: Comparison of MammaPrint, BluePrint, and TargetPrint with Clinical Parameters in Patients with Breast Cancer: Findings from a Prospective United States Cohort.
Authors: Nguyen B , Sinha R , Kerlin D , Barone J , Garcia A , Yao K , Rivera E , Stork-Sloots L , Deck K
Published: June 2012

Background: MammaPrint (MP) is a powerful predictor of disease outcome in early stage breast cancer. In addition, TargetPrint (TP), a quality approved microarray-based test that measures the mRNA expression level of ER, PR and HER2 and an 80 gene expression Molecular Subtyping profile BluePrint (BP) were developed. In the present study, MP, BP and TP were measured in a prospective U.S. breast cancer patient cohort.

Methods: MP results were evaluated in fresh tumor samples from 141 breast cancer patients (T1-4N0-2; median age 62 [35-97 yr]) collected by core needle biopsy or from a surgical specimen between July 2008 and February 2011. We compared treatment advice as recommended by NCCN guidelines and classification according to MP. In addition, we compared IHC/FISHER, PR and HER2 assessments with TP. The MP and BP results were used to subtype the patients into molecular subgroups.

Results: For the group of patients (n=69) for which NCCN recommends the use of a multi-gene signature for determining chemotherapy treatment recommendations, 50 patients were classified as High Risk and 19 as Low Risk by MP. Comparison of TP with IHC/FISH indicated a concordance of 98% for ER, 91% for PR, and 94% for HER2. For a subgroup of 63 patients combined MP and BP results were available; 22 patients were Luminal-type/MP Low Risk, 31 patients were Luminal-type/MP High risk, 1 patient was Her2-type/MP Low Risk, 2 patients were Her2-type/MP High Risk and 7 patients were Basal-type/MP High Risk.

Conclusion:The multi-gene signature MammaPrint, as well as BluePrint and TargetPrint provides additional information for treatment guidance. By combining MammaPrint with the BluePrint molecular subtyping profile, specific groups of patients can be recognized that are at high risk of recurrence and that would possibly benefit from specific treatment. This study shows that TargetPrint provides high quality second opinion for local IHC/FISH assessment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-06.
P3-04-07: Physiological Concentrations of Genistein and 17b-Estradiol Inhibit MDA-MB-231 Breast Cancer Cell Proliferation by Increasing Bax/Bcl2 Ratio and Decreasing pERK1/2 Expression.
Authors: Rajah TT , Peine KJ , Du N , Serret CA
Published: June 2012

Background: Our previous results were the first to show that physiological concentrations (1 μM) of genistein, a soy component, in the presence of17β-estradiol (1 nM) inhibited the cell proliferation of MDA-MB-231 (Estrogen receptor (ER) β positive) breast cancer cells. These results are relevant in premenopausal women with breast cancer of the ERα-negative and ERβ-positive type, especially given the increasing trends of soy intake among the US population in the past decade. The aim of the present study was to identify the mechanism by which genistein plus 17β-estradiol inhibits the cell proliferation of ERα-negative and ERb-positive breast cancer cells. Our hypothesis is that the balance of signaling actions by genistein plus 17β-estradiol from different signaling pathways is likely to lead to the cell's choice to either proliferate or enter the apoptotic pathway. For this purpose, the effect of low and high concentrations of genistein (1 μM and 100 μM) in the presence or absence of 17β-estradiol (1 nM) was studied on the expression of cell signaling proteins involved in cell proliferation, survival and apoptosis (pERK1/2, pAkt, Bax and Bcl2) and correlated to cell proliferation and apoptosis in MDA-MB-231 (ERβ positive and ERα negative) breast cancer cells.

Methods: Cell proliferation was determined by the MTT assay, apoptosis determined microscopically by the use of acridine orange and ethidium bromide dyes and the expression of cell signaling proteins by western blotting.

Results: Our results show that 1μM genistein plus 17β-estradiol significantly increased apoptosis (p<0.05) as compared to the control (12.47% vs 5.87% respectively). Increased expression of Bax/Bcl2 (2.5 fold) along with a decreased expression of phosphorylated ERK1/2 (2 fold) was observed in cells treated with 1μM genistein plus 17β-estradiol as compared to the control cells. Phosphorylated Akt did not show any differences in the treatment condition as compared to the control. High concentrations of genistein (100 μM) in the presence or absence of 17β-estradiol also increased apoptosis; however these changes could not be correlated to the expression of Bax/Bcl2, or pERK1/2.

Conclusion: In conclusion, our results show that physiological concentrations of genistein in the presence of17β-estradiol inhibit cell growth through apoptosis via increased Bax/Bcl2 and a concomitant decrease in pERK1/2 expression. Our results also suggest that different concentrations of genistein elicit cell responses through different signaling mechanisms. These results are especially relevant to the cohort of premenopausal women with breast cancer of the ERβ positive and ERα negative type.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-07.
P3-06-03: Hypodiploidy, 1pter Loss and Inactive X Chromosome Retention Are Associated with BRCA1 Somatic or Germline Inactivation in Basal-Like Breast Carcinomas: Proposal for a New BRCAness Genomic Signature.
Authors: Manié E , Popova T , Vincent-Salomon A , Dubois T , Delattre O , Sastre-Garau X , Stoppa-Lyonnet D , Stern M-H
Published: June 2012

Basal-Like Carcinomas (BLCs) are high grade ductal carcinomas identified by large scale transcriptomic analyses among carcinomas with triple-negative phenotype (ER- PR- HER2−). A particular relationship between this phenotype and the BRCA1 breast cancer susceptibility gene has been described based on several observations including high genomic similarities of sporadic and BRCA1 BLCs. BRCA1 mutated patients have been shown to strongly benefit from treatments using PARP inhibitors. However, significant benefit of this treatment was not demonstrated in sporadic triple-negative breast cancers in association to standard chemotherapy in phase III clinical trials, which emphasizes the necessity for a better selection of DNA repair deficient tumors. The goal of this study was to determine whether BRCA1 inactivation leads to particular genomic alterations that could be used to identify BRCA1 deficient sporadic BLCs.

Genomic profiling was performed using SNP-arrays (Affymetrix and Illumina) and high quality profiles were obtained for 60 BLCs with more than 35% of tumor cells. The series contained 28 BRCA1 BLCs and 32 sporadic BLCs consisting in 10 sporadic BLCs with BRCA1 somatic inactivation by promoter methylation (BRCA1-like BLCs) and 22 sporadic tumors without BRCA1 methylation (nonBRCA1 BLCs). Genomic data was mined with GAP methodology (Popova et al, Genome Biol 2009), which allows absolute copy-number evaluation with DNA content calculation and clarification of Loss Of Heterozygosity (LOH) status.

Genomic patterns of BLCs were characterized by frequent and highly recurrent allelic losses. Comparison between BRCA1, BRCA1-like and nonBRCA1 BLCs confirmed their overall similarity, and identified few significant differences enhanced when BRCA1 and BRCA1-like BLCs were considered as one group. Firstly, hypodiploidy characterized BLCs with BRCA1 germline and/or somatic inactivation. Secondly, the lpter region was found more often lost in BRCA1 and BRCA1-like BLCs than in nonBRCA1 BLCs. Thirdly, retention of inactive X chromosome was a characteristic of BLCs with BRCA1 germline and/or somatic inactivation. Finally, a BRCA1ness score based on the above described difference is proposed.

In conclusion, despite a strong similarity of genomic patterns in BLCs, BRCA1 inactivation leads to few key genomic aberrations which constitute a new BRCA1 molecular signature namely hypodiploidy, 1pter loss and inactive X chromosome retention. This signature needs to be further evaluated in terms of response to PARP inhibitors in order to evaluate its capacity to identify patients that will benefit from this treatment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-06-03.
P3-07-07: Effect of Sentinel Lymph Node Biopsy without Axillary Lymph Node Dissection on Overall Survival in Patients with T1 or T2 Node-Positive Breast Cancer: A Report from the Korean Breast Cancer Society.
Authors: Park HS , Chae BJ , Song BJ , Jung SS
Published: June 2012

Introduction: Sentinel lymph node biopsy (SLNB) is accepted as an alternative method to axillary lymph node dissection for staging axillary lymph node status in clinically node-negative breast cancer. Current practical guidelines recommend that axillary lymph node dissection (ALND) should be performed in cases with sentinel node metastasis, and most of node-positive patients should receive adjuvant systemic therapy to reduce locoregional/distant recurrence and to improve overall survival irrespective of the number of lymph node metastasis. However, patients with ALND are more likely to develop lymphedema than those with SLNB alone, and appropriate systemic chemotherapy or hormone therapy significantly reduce locoregional and distant recurrence in early breast cancer patients. For this reason, the previous prospective study, American College of Surgeon Oncology Group Z0011 trial, was conducted and it suggested that there is no difference in overall survival between node-positive patients who received breast conserving treatment with SLNB alone and those with ALND after SLNB. This study is aimed to evaluate the difference of survival between node-positive patients who underwent SLNB alone and those who received ALND after SLNB using the Korean Breast Cancer Society registry.

Methods: In 87671 patients with breast cancer in the registry, we enrolled 2581 patients who meet the eligible criteria in the study. All enrolled patients had T1 or T2 breast cancer, and received mastectomy or breast conserving treatment followed by documented adjuvant systemic therapy between Jan. 2001 and Apr. 2011. Log-rank test and Cox-proportional hazard model were used to access the difference of overall survival according to the axillary procedure.

Results: There were 197 patients with SLNB alone and 2384 patients with ALND after SLNB, respectively. Smaller tumor size, lower number of nodal metastasis, and higher proportion of breast conserving surgery were shown in patients with SLNB alone than in those with ALND after SLNB. There was no significant difference in overall survival between 2 groups in the log-rank test. ALND after SLNB showed no significant improvement on overall survival in Cox-propotional hazard model adjusted by tumor size, number of nodal metastasis, and operation type (P=0.78, HR=0.73, 95% CI=0.08−6.62).

Conclusion: The current study suggests that ALND after SLNB in cases with sentinel lymph node metastasis may not influence on the improvement of overall survival and supports the results of Z0011 trial. Further validation studies are necessary to expand the understanding of the role of performing SLNB alone in patients with node-positive breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-07.
P3-07-19: Long-Term Outcome of Internal Mammary Lymph Node Detected by Lymphoscintigraphy in Early Breast Cancer.
Authors: Koo MY , Lee JE , Nam SJ , Yang J-H , Lee SK , Kim S , Choi M-Y , Cho DH , Kim J , Lee J , Jung SP
Published: June 2012

Purpose: The purpose of this study is to determine the long-term significance of internal mammary lymph node (IMLN) detected by lymphoscintigraphy.

Background

IMLN metastasis is an important prognostic indicator in breast cancer. However, the necessity of internal mammary sentinel lymph node (IMSLN) biopsy for accurate staging, for choosing adjuvant treatment, and as a prognostic indicator, has remained controversial.

Methods: From January 2001 until December 2006, 525 female breast cancer patients underwent radical surgery after preoperative lymphatic scintigraphy. We retrospectively analyzed the follow-up results, recurrences and deaths of all patients.

Results: There was no significant difference in clinicopathologic characteristics between the axilla and the IMLN group. The median follow-up period was 118.8 (range, 7–122) months in the axilla group and 107.7 (range, 14–108) months in the IMLN group. During the median follow-up period, the breast cancer-related death rate in the axilla group was 3.6%, which was not significantly different from that of the IMLN group (1.3%) (P = 0.484). The five-year survival rates (5YRS) did not differ between the two groups (P = 0.306). The overall recurrence rate and the locoregional (LR) recurrence rate also did not differ between the two groups (P = 0.835 and P = 0.582, respectively. The recurrence rate of IMLN (both ipsilateral and contralateral) metastasis was very low, accounting for 0.5% in the axilla group and 1.3% in the IMLN group (P = 0.416).

Conclusion: The long-term follow up results showed that there was no significant difference in both overall outcome and regional recurrence between the two groups. Therefore, the necessity of identification of nodal basins outside the axilla or the necessity of IMLN sentinel biopsy should be reconsidered.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-19.
P3-07-29: Validating the Lymph Node Ratio as a Prognostic Indicator among South East Asian Breast Cancer Patients.
Authors: Saxena N , Hartman M , Bhoo Pathy N , Aziz R , Siew EL , Lee SC , Yip CH , Verkooijen HM
Published: June 2012

Background: Several studies in Caucasian settings have suggested that the lymph node ratio (LNR, i.e. the ratio of the number of positive nodes to the total number of nodes excised) is a superior prognostic indicator for breast cancer as compared to the absolute number of nodes involved (pN stage). This study validates the prognostic performance of LNR in the South East Asian setting.

Methods: All patients diagnosed with non-metastatic invasive breast cancer at the National University Hospital (Singapore) or University of Malaya Medical Center (Kuala Lumpur) between 1990 and 2007 and with information on axillary nodes removed and involved were included for analysis (n = 1589). Multivariate Cox regression analysis was performed to evaluate the prognostic value of the LNR [low (> 0 and <0.2), intermediate (0.2 to 0.65) and high risk (>0.65 to 1)] and pN staging [pN1, pN2 and pN3] for all cause mortality. Patients staged pN0/LNR 0 were excluded from analysis.

Results: According to the LNR classification, 758 patients were categorized as low risk, 574 as intermediate risk and 257 as high risk LNR. For classic pN staging, 879 were pN1, 447 pN2 and 263 pN3. Women in the intermediate risk category (0.2 to 0.65) had a 1.5 fold increased risk of death [Adjusted hazard ratio [HRadj] of 1.5 (95%CI, 1.2 to 1.9]), and women with high risk (>0.65 to 1) LNR had a 3.2 fold increased risk of death [HRadj of 3.2 (95%CI, 2.6 to 4.1)] as compared to the low risk women (>0 and <0.2). Similarly, women with pN2 disease had a 1.9 fold increased risk of death [HRadj of 1.9 (95%CI, 1.5 to 2.4)], and women with pN3 disease had 3.1 fold increased risk of death [HRadj 3.1 (95%CI, 2.5 to 3.9)] compared to pN1 patients.

Conclusion: Among South East Asian breast cancer patients, both the Lymph Node Ratio and the pN Staging system seem to be equally good at predicting all cause mortality.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-29.
P3-07-46: Accuracy of SPIO-Enhanced MR Imaging Alone for the Diagnosis of Sentinel Node Metastases in Patients with Breast Cancer.
Authors: Motomura K , Nakahara S , Ishitobi M , Komoike Y , Koyama H , Inaji H , Horinouchi T , Nakanishi K
Published: June 2012

Background

Superparamagnetic iron oxide (SPIO)-enhanced MR imaging has been reported to be promising for the detection of metastases in sentinel nodes localized by CT lymphography in patients with breast cancer (Motomura, Ann Surg Oncol 2011). The current SPIO technique involves imaging before and after contrast administration. This study evaluated the accuracy of SPIO-enhanced MR imaging alone without unenhanced imaging.

Methods: This study included 120 patients with breast cancer demonstrating clinically negative nodes. Sentinel nodes were identified by CT lymphography, and MR imaging of the axilla before and 18–24 hr after interstitial administration of SPIO was performed. A node was considered non-metastatic if it showed a homogenous low signal intensity and metastatic if there was an absence of low signal intensity either in the entire node or in a focal area on SPIO-enhanced MR imaging. The diagnostic accuracy of the SPIO-enhanced imaging alone was compared with that of combined unenhanced and SPIO-enhanced imaging.

Results: The mean number of sentinel nodes identified by CT lymphography was 1.2 (range 1–3). Pathologic evaluation demonstrated that 28 (23.3%) of 120 patients showed metastasis to at least one node. One false negative result was added when the evaluation was based solely on SPIO-enhanced MR imaging. Consequently, the sensitivity decreased from 84.0% to 80.0% and the accuracy decreased from 89.2% to 88.3%, respectively, in the SPIO-enhanced MR imaging alone. However, the differences in sensitivity and accuracy between SPIO-enhanced MR imaging alone and the combined unenhanced and SPIO-enhanced imaging were not significant (McNemar's test; p=1.0). The specificity of enhanced imaging alone and that of combined unenhanced and SPIO-enhanced imaging were both 90.5%.

Conclusions: A single MR imaging examination performed after SPIO administration can be used for accurate diagnosis of sentinel node metastases, and thus reduce the time and cost of imaging.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-46.
P3-07-47: Validated Nomogram To Predict Sentinel Lymph Node Positivity in Breast Cancer Patients.
Authors: Kaplan R , Qiu Q , Monni S , D'Alfonso T , Shin SJ
Published: June 2012

Background: Sentinel lymph node(SLN) biopsy is commonly used to assess axillary lymph node status in breast cancer patients. Intraoperative (frozen section) evaluation of SLN may lead to immediate axillary dissection if the lymph node is found to be positive. In light of the recent outcomes of the Z00011 trial, the use of intraoperative evaluation for SLN should diminish since only select patients appear to benefit from axillary dissection. We have previously reported that tumor size and lymphovascular invasion (LVI) strongly predict SLN positivity in a cohort of 350 breast cancer patients [Modern Pathology (2011) 24, 30A]. From this test cohort, we developed a nomogram which could help identify patients more likely to have sentinel lymph node positivity. The aim of this study was to validate the efficacy of this nomogram by applying a new, non-overlapping cohort of breast cancer patients who had undergone sentinel lymph node sampling. Materials and Methods: 93 breast cancer patients aged 29–82 (mean:59) who underwent excision (54, 58%) or mastectomy (39, 42%) with a SLN biopsy were studied. Patients who had undergone neoadjuvant chemotherapy were excluded. Clinicopathologic parameters such as number of sentinel lymph nodes removed and/or positive, number of non-sentinel lymph nodes removed and/or positive, tumor size, histologic type/grade, presence of lymphovascular invasion, biomarker expression (ER,PR, HER-2/neu) and multifocality were recorded. Statistical analysis was performed to identify which variables correlated with SLN positivity in this validation cohort. In addition, the probability of SLN positivity for each case was determined by using the nomogram and this value was then compared to the patient's true SLN status. Results: When analyzed separately, tumor size (p=0.03) and LVI (p=0.01) were variables that were significantly associated with SLN positivity in both the test and validation cohorts. Employment of the nomogram demonstrated that 60% of patients who were predicted to have a positive SLN with at least 0.8 probability were truly positive by histologic and immunohistochemical examination. Moreover, 89% of patients who were predicted to have a negative SLN with at least 0.8 probability were truly negative by histologic and immunohistochemical examination. Conclusions: We conclude that the utility of this nomogram prior to surgery can help predict SLN positivity. This can serve as a complimentary adjunctive tool to better select patients who will likely need intraoperative evaluation of their sentinel lymph node due to their higher risk of sentinel node positivity and thus, benefit from the option of immediate axillary dissection at that time.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-47.
P3-09-06: Changes of Serum Vitamin D According to the Breast Cancer Treatment.
Authors: Kim HJ , Yi OV , Koh BS , Yu JH , Lee JW , Son BH , Ahn SH
Published: June 2012

Background: Vitamin D deficiency is associated with increased breast cancer risk and decreased breast cancer survival. The purpose of this study was to determine the effect of breast cancer adjuvant treatment to the vitamin D status, as measured by the serum hydroxyvitamin D (25OHD) in breast cancer patients.

Patients and Methods: For 589 patients who was diagnosed as a non metastatic breast cancer in 2009 at the asan medical center, blood was prospectively analyzed in batches for serum 25 OHD level at basal and at 6 and 12month. We excluded the patients who took a vitamin D supplementation and got a neoadjuvant chemotherapy. Vitamin D sufficiency was defined as serum as 30ng/ml or greater, insufficiency as 20 to 29 ng/ml and insufficiency as less than 20ng/ml.

Results: At baseline, mean serum 25OHD was greater in summer (April to Oct) than Winter(Nov to May) (28.2ng/ml vs 32.9ng/ml respectively, p=0.000). The patients who did not get a chemotherapy and antihormonal therapy as baseline, the patient with chemotherapy showed decreased serum 25OHD level than who without chemotherapy in 6 month but not in 12 month (p=0.003, vs p=0.156 respectively). The patients who had taken anti-hormonal therapy showed significant increasing serum 25OHD in 6 month and 12 months (p=0.000 both). For the patients who got both chemotherapy and anti-hormonal therapy, the changes of serum 25OHD level is smaller than the patients who got a chemotherapy only.

For the patients who got a chemotherapy, 57% of patients were vitamin D sufficient at baseline, but 27% of patients in 6 month and 49% in 12 month (p=0.001).

Conclusion: Vitamin D status was worsen during chemotherapy but recovered after chemotherapy. Anti hormonal therapy make the serum vitamin D level increased. The translational research about the effect of chemotherapy and anti-hormonal therapy to the vitamin D status should be warranted.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-09-06.
P3-10-04: Halting Early Breast Cancer Progression with Omega-3 Ethyl Esters: Altering Tumor Microenvironment in 21T Series Cell Lines.
Authors: Chen CH , Rhodes ME , Fabian C , Hursting SD , deGraffenried LA
Published: June 2012

Observable infiltration of immune cells in stages of early breast cancer progression mirrors two recently conceptualized factors of carcinogenesis: tumor microenvironment and tumor-promoting inflammation.

Clinical studies using immunohistochemical staining for CD45 in patient-derived breast tissue demonstrate the involvement of leukocytes in breast cancer development, but the lack of an appropriate in vitro model has hindered molecular studies thus far. However, the 21T series cell lines effectively mimic breast cancer progression. This cell line series was obtained from the same breast cancer patient and consists of three cell lines representing atypical hyperplasia (21PT), ductal carcinoma in situ (21NT) and invasive carcinoma (21MT-1). This unique cell line series is used in our molecular study of omega-3 ethyl esters as breast cancer preventive agents. Preliminary invasion assays have described varying degrees of monocyte recruitment according to stages of progression. 21PT cells when plated on the bottom chamber failed to mobilize monocytes as effectively as 21NT or 21MT-1 and the addition of omega-3 ethyl esters suppressed the baseline invasion capacity of monocytes by 30%. Epidemiologic studies suggest that omega-3 fatty acids can act as potent cancer preventive agents and are possibly more potent when used in an ethyl ester form. Based on this, we hypothesize that omega-3 ethyl esters may prohibit early breast cancer progression through inhibiting immune cell infiltration and disrupting the formation of a tumor-promoting inflammatory and oxidative microenvironment. Phenotypic studies aim to interrogate the effect of omega-3 ethyl esters on tumor microenvironment including chemokine secretion and MMP production whereas molecular experiments probe into the importance of the Wnt pathway as a key player in the transition from ADH to DCIS. This study will demonstrate how omega-3 ethyl esters may function as breast cancer preventive agents as well as provides a model of study to test the efficacy of cancer preventive agents in vitro.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-10-04.
P3-12-05: Breast Cancer Recurrence: 2nd Conservative Treatment Versus Mastectomy.
Authors: Michel V , Houvenaeghel G , Bannier M , Tallet A , Minsat M , Lambaudie E , Salem N , Butarelli M , Resbeut M
Published: June 2012

Objectives: Mastectomy (Mt) is considered standard treatment for isolated local recurrence (LR) of breast carcinoma. The aim of our study was to evaluate a second conservative treatment (defined as lumpectomy followed by interstitial brachytherapy (LpIB)) and to determine if it compares favorably with the current standard treatment. Materials and methods: Between January 1981 and December 2009, 348 patients were treated to the Paoli Calmettes Institute (IPC) for an isolated LR: 232 (66,7 %) underwent Mt, 62 (17,8 %) received a second radio-surgical conservative treatment (LpIB) and 54 (15,5 %) a 2nd single surgical treatment (Lp).

We classified each population according to the well known prognosis factors.

Then, populations Mt and LpIB were matched taking into account these criteria to compare the overall survival (OS), metastasis free survival (MFS) and 2nd local recurrence free survival according to the treatment delivered.

Results: On 348 reviewed patients, with a median follow-up of 73,3 months, 65 patients died (42/232 Mt, 8/62 TecCur, 15/54 Tec) and 100 presented metastasis (64/232 Mt, 15/62 TecCur, 21/54 Tec). There was no difference in MFS for the 2 groups, LpIB and Mt (80 % at 5 years) and the OS was non significantly better in the group LpIB compared to the group Mt (90 % and 82 % at 5 years respectively, p=0,28), whereas in the LpIB group 17% and 30% presented a relapse at 5 years and 10 years respectively.

They subsequently underwent a salvage mastectomy.

Worse results were obtained with lumpectomy alone (OS = 72 % and MFS = 68 % at 5 years) compared with 2 other option treatments.

Conclusion: A second conservative treatment for breast cancer recurrence, i.e. lumpectomy and interstitial brachytherapy, is possible for selected patients, without any negative impact on overall survival, nor metastasis free survival.

Keywords: Local recurrence, interstitial brachytherapy, lumpectomy, conservative treatment, mastectomy, overall survival, metastasis free survival, breast carcinoma.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-12-05.
P3-13-03: Long-Term Symptoms after Radiotherapy of Supraclavicular Lymph Nodes in Breast Cancer Patients.
Authors: Lundstedt D , Gustafsson M , Steineck G , Alsadius D , Sundberg A , Wilderäng U , Holmberg E , Johansson K-A , Karlsson P
Published: June 2012

Background and Purpose: Irradiation of the supraclavicular lymph nodes has historically been shown to increase the risk of brachial plexopathy with neurological problems in the upper limb. The purpose of this study was to compare long-term symptoms after modern radiotherapy (based on 3D dose planning) in breast cancer patients with or without irradiation of the supraclavicular lymph nodes.

Material and Methods: We collected information from 814 recurrence free women consecutively treated with adjuvant radiotherapy for breast cancer at the Sahlgrenska University Hospital in Gothenburg, Sweden, 1999 to 2004. The women had breast conserving surgery or mastectomy with axillary dissection or sentinel node biopsy. The breast area was irradiated to 50 Gy in 2.0 Gy fractions. Women with more than three lymph node metastases had regional radiotherapy to the supraclavicular lymph nodes delivered in 2.0 Gy fractions up to 50 Gy. Systemic treatments were given according to regional guidelines. In this study the women were classified into three groups depending on if they had axillary dissection and regional radiotherapy. The first group had both axillary dissection and regional radiotherapy, the second group had axillary dissection without regional radiotherapy, and the third group had sentinel node biopsy (i.e. no axillary dissection) without regional radiotherapy. Three to eight years after radiotherapy, the women received a questionnaire asking about paresthesia, pain and strength in the upper limb.

Results: Among women with axillary dissection and regional radiotherapy 38/192 (19.8%) reported paresthesia in the hand compared to 68/505 (13.5%) among women with axillary dissection without regional radiotherapy; relative risk (RR) 1.47; 95% confidence interval (95% CI) 1.02 - 2.11, and compared to 9/112 (8.0%) among women with sentinel node biopsy without regional radiotherapy; RR 2.46 (95% CI 1.24−4.90). Type of breast surgery, number of examined axillary lymph nodes, and chemotherapy had no impact on the occurrence of paresthesia. Age was an effect modifier among the women with axillary dissection and regional radiotherapy; up to 49 years of age 26.8% reported paresthesia (RR 2.45; 95% CI 1.05−5.73), between 50 and 59 years of age 19.7% reported paresthesia (RR 1.81; 95% CI 0.73−4.44), and above 59 years of age 10.9% reported paresthesia (RR 1.00 Reference). We found no statistically significant differences between the groups regarding pain or decreased strength.

Conclusions: Radiotherapy to the supraclavicular lymph nodes increases the occurrence of paresthesia in the hand. The effect was mainly seen among younger women. Dose/volume-response analysis regarding paresthesia will be presented at the meeting.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-13-03.
P3-13-10: Does Lapatinip Increase Pulmonary Toxicity When Concurrently Used with Radiation Therapy? An Experimental Study with Wistar-Albino Rats.
Authors: Senel Bese N , Yetmen O , Guzel E , Karaca C , Coban I , Elicin O , Suzer O
Published: June 2012

Purpose: Lapatinib (L) is an oral receptor tyrosine kinase inhibitor which has shown activity in the treatment of metastatic breast cancer. Adjuvant usage of L is being investigated in clinical phase III trials. There is no data regarding the side effects of combination of RT and L which may be a problem when L is used in the adjuvant setting. Lung is the most radiosensitive organ to observe late effects of RT. We evaluated if concurrent administration of L has any impact for the development of radiation induced pulmonary fibrosis in rats (RIPF). Material Methods: 40 female Wistar-albino rats (WAR) were divided into 4 experimental groups (G). G1 (control) did not receive any treatment. G2 (RT) received RT to whole thoracic region. G3 (L) received L without RT. G4 (L+RT) received L with RT. A total dose of 30 Gy in 10 fractions was given to both lungs with an anterior field at 2 cm depth. L equivalent to 1500 mg/day, 60 kg adult dose, were calculated according to the mean weight of rats, orally administrated with a feeding tube twice daily including the week-ends until WAR were sacrificed. WAR were anesthetized and sacrificed 16 weeks after RT which was shown to be a sufficient period for the development of RIPF in rats. Paraffin sections (5 μm thick) of lungs were stained with hematoxylin-eosin and Masson's trichrome. A comparative analysis was performed among 4 groups by scoring the pulmonary injury between 0 and 3 according to the infiltration of inflammatory cells into the alveolar spaces, alveolar wall thickening and architectural deformation across the entire lung section. Normality distribution were tested, then one-way ANOVA followed by post hoc Holm-Sidak testing were used.

Results: In G2 inflammatory cell infiltration, fibrosis with damage to lung structure and formation of small fibrous masses were observed. Alveolar septa was significantly thicker than G1 (p<0.05), which revealed totally normal pulmoner structure. G3 showed minimal alveolar septal thickening and infiltration of inflammatory cells into the alveolar spaces which was not significantly different than G1. Histopathological findings in G4 were similar to those in G2 and statistically different when compared with the G1 and G3(p<0.05). Conclusion: Study shows that addition of L to RT does not increase RIPF in rats.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-13-10.
P3-14-19: Impact of Chemotherapy-Induced Amenorrhea on Response to Neoadjuvant Chemotherapy in Breast Cancer.
Authors: Ahn SK , Moon HG , Kim JS , You JM , Shin HC , Han W , Noh D-Y
Published: June 2012

BACKGROUND: Although chemotherapy and ovarian ablation independently improve the outcome of breast cancer, there is controversy about the benefit of chemotherapy-induced amenorrhea (CIA) in breast cancer. We investigated impact of CIA on response to neoadjuvant chemotherapy in breast cancer patients.

METHODS: We reviewed the records of 198 premenopausal patients with breast cancer treated with neoadjuvant chemotherapy between January 2005 and December 2010. Chemotherapy-induced amenorrhea (CIA) was defined as serum FSH level ≥40 IU/L after completion of all scheduled neoadjuvant chemotherapy and prior to definitive surgery.

RESULTS: Among 198 breast cancer patients, 132 pts (66.7%) developed CIA after neoadjuvant chemotherapy. 156 pts (78%) underwent DA chemotherapy. The age of CIA patients was older than non-CIA patients (41.55±5.55 years vs. 38.27 ± 6.86 years, p=0.001). The incidence of CIA after neoadjuvant chemotherapy was significantly higher in responder group (responder vs. nonresponder: 87 pts (74.4%) vs. 45 pts (55.6%); p=0.006). Additionally, FSH level after all scheduled neoadjuvant chemotherapy was significantly higher in responder group (FSH 56.41±32.41 IU/L vs. 45.76±30.31 IU/L; p=0.021). In univariate analysis, CIA (p=0.006) and total number of chemotherapy cycle regardless of chemotherapy regimen (p=0.04) were significantly associated with tumor response. CIA was only independent factor for tumor response after neoadjuvant chemotherapy on multivariate analysis (p=0.012).

CONCLUSION: CIA after neoadjuvant chemotherapy was significantly associated with response to neoadjuvant chemotherapy in locally advanced breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-19.