bluuebunny/crossref_metadata_2025 huggingface dataset:
P2-11-09: EGFR Overexpression in Triple Negative Breast Cancer (TNBC) and Its Association with the Prognosis.
Authors: Liu W , Zhang L , Ma K , Han B , Li S , Xu G , Fan Z , Liu N , Shi A
Published: June 2012

Objective: The aim of this study is to investigate EGFR expression in Triple Negative Breast Cancer (TNBC), and to find the relationship between EGFR overexpression and prognosis of TNBC, further to clarify the significance of EGFR in TNBC and provide valuable information for TNBC therapy.

Methods: 42 triple ***negandection ***ssitive breast cancer patients(studying group) and 40 HER2(3+) breast cancer patients(controling group) who underwent surgery from January 2000 to December 2005 were analyzed. 82 cases of paraffin-embedded specimens were detected by Immunohistochemistry(IHC), fluorescence in situ hybridization(FISH) and polymerase chain reaction(PCR) to investigate the overexpression, amplification and mutation of EGFR gene. The distant-free survival(DFS) and overall survival(OS) of these patients were used to investigate the relationship between EGFR overexpression and the prognosis of TNBC.

Results: 34(43.9%) EGFR overexpression was observed in all cases, while gene amplification was only 7(9.1%) cases. No EGFR gene mutation was found in all cases. Overexpression of EGFR occurring in 57.1% patients in TNBC group and 25.0% patients in HER2 group, and we didn't found any correlation between EGFR overexpression and clinicopathology. 50(61.0%) patients relapsed (TNBC 28,HER2 22) and 27(32.9%) patients died(TNBC 18,HER2 9) were observed during the more than 5 years follow-up. The 5-year DFS was 57.1% and 77.5% respectively, the 5-year OS was 71.4% and 87.5% in TNBC and HER2 groups. In TNBC group, the survival of the EGFR-overexpressing group patients was significantly lower than that of the non-EGFR-overexpressing group patients (p=0.018 for DFS, p=0.026 for OS); In HER2 group, no statistical difference was found (p=0.079 for DFS, p=0.055 for OS).

Conclusions: This study showed that EGFR overexpression increased significantly in TNBC patients, which was no correlation with their clinico-pathological data. EGFR gene amplification was much less frequent than its overexpression. It suggested that EGFR gene amplification may not be the unique mechanism of EGFR overexpression in TNBC. There may be other possible mechanisms and pathways that cause EGFR overexpression. In addition, it may suggest that gefitinib therapy is useless in TNBC patients because we did not find any mutations in the tested exons of TNBC. EGFR overexpression may associate with a poor outcome of TNBC patients which suggest it could be a significant prognostic factor for TNBC patients. EGFR may play important role for molecular-targeting therapy of TNBC.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-11-09.
P2-12-05: Correlation between Quantitative HER2 Protein Expression and Risk of Brain Metastases in HER2−Positive Advanced Breast Cancer Patients Receiving Trastuzumab-Containing Therapy.
Authors: Duchnowska R , Biernat W , Szostakiewicz B , Sperinde J , Piette F , Haddad M , Paquet A , Lie Y , Czartoryska-Arlukowicz B , Wysocki P , Jankowski T , Radecka B , Foszczynska-Kloda M , Litwiniuk M , Debska S , Weidler J , Huang W , Buyse M , Bates M , Jassem J
Published: June 2012

Background. Patients with HER2−positive breast cancer are at particularly high risk for brain metastases; however, the biological basis is not fully understood. Within HER2−positive breast cancer tumors, it is possible to resolve a ∼1.5-log range of HER2 protein expression using a novel quantitative HER2 assay (HERmark®). We investigated the correlation between quantitative HER2 protein expression in primary breast cancers and the time to brain metastases (TTBM) in HER2−positive advanced breast cancer patients treated with trastuzumab.

Methods. The study group included 142 consecutive patients who were administered trastuzumab-based therapy for HER2−positive metastatic breast cancer, defined as 3+ categorical staining by immunohistochemistry (IHC). HER-2/neu gene copy number was subsequently quantified as HER2/CEP17 ratio by central laboratory fluorescence in situ hybridization (FISH). HER2 protein was quantified as total HER2 protein expression (H2T) by the HERmark assay in formalin-fixed, paraffin-embedded primary tumor samples. HER2 variables were correlated with clinical features and TTBM measured from the initiation of trastuzumab-containing therapy.

Results. H2T level (continuous variable) was correlated with shorter TTBM (HR=2.3; p=0.013), whereas HER2 gene amplification by FISH (p=0.28) and continuous HER2/CEP17 ratio (p=0.25) had no significant prognostic impact. The correlation between continuous H2T level and TTBM was confirmed in a multivariate analysis (HR=3.2; p=0.021). Controlling for the competing risk of death from causes other than brain metastases, continuous H2T remained a strong correlate of TTBM (HR=2.7; p=0.0009). In the subset of patients that was centrally-determined HER2 positive by FISH (117 patients), above-median H2T level was significantly associated with shorter TTBM (HR=2.4; p=0.005), whereas this was not true for median FISH/CEP17 ratio (p=0.4). In a multivariate analysis of this subset, continuous H2T (p=0.021) and a time dependent covariate capturing time to non-brain metastases (p=0.0044) were prognostic for TTBM, whereas FISH/CEP17, ER, PgR and grade were not.

Conclusions. These data reveal a strong relationship between quantitative HER2 protein expression levels and the risk of brain relapse in HER2−positive advanced breast cancer patients. Consequently, quantitative assessment of HER2 protein expression may inform and facilitate refinements in therapeutic treatment strategies for selected subpopulations of patients in this group.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-05.
P2-12-06: Nomogram To Predict Subsequent Bone Metastasis in Patients with Non Metastatic Breast Carcinomas.
Authors: Lousquy R , Delpech Y , Rouzier R , Gligorov J , Hsu L , Barranger E , Pusztai L , Uzan S , Hortobagyi GN , Coutant C , Ibrahim NK
Published: June 2012

Background

Bone metastasis (BM) is one of the most common sites of distant metastasis for breast cancer. We hypothesized that its occurrence can be predicted if a clinical nomogram can be developed, thus allowing for selection of patients at high risk for BM.

Patients and Methods

Medical records of patients with non metastatic breast cancer were prospectively collected for the period between January 1997 and February 2007 at the M.D. Anderson Cancer Center (Texas, USA). A multivariate logistic regression analysis of selected prognostic features was done. Based on Cox proportional hazards regression model, a nomogram to predict BM was constructed and validated in an independent cohort of 579 patients with non metastatic breast cancer at time of diagnosis treated at the Tenon Hospital (Paris, France) between January 2003 and December 2005.

Results: Of 4175 patients with non metastatic breast cancer, 314 developed subsequent BM. Age, T stage, lymph node status, lymphovascular space invasion, breast cancer molecular subtype, adjuvant hormonotherapy were significantly and independently associated with subsequent BM. The nomogram had a concordance index of 0.69 (95% CI, 0.68 to 0.70) in the training set. The validation set showed a good discrimination with a concordance index of 0.65 (95% CI, 0.57 to 0.72). At 3, 5 and 7 years, the nomogram was well calibrated.

Conclusion: We have developed a robust tool that is able to predict subsequent BM in patients with non metastatic breast cancer. Selection of an enriched patient population at high risk for BM will allow to practice individualized therapeutic strategies, an adapted medical supervision and will facilitate the design of trials aiming at its prevention with the use of biphosphonate treatment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-06.
P2-12-14: Prognostic Value of HER2 Positivity and Negative Hormonal Status in Patients with Small Tumor (<1cm) and Node-Negative Breast Cancer.
Authors: Meattini I , Livi L , Saieva C , Agresti B , Scotti V , Nori J , Sanchez LJ , Vezzosi V , Bianchi S , Cataliotti L , Biti G
Published: June 2012

Introduction. Human epidermal growth factor receptor 2 (HER2) amplification has become the prototype biomarker to develop tailored biological treatment. Many studies suggested that HER2 positivity is an independent predictor of disease recurrence and breast cancer mortality. Trastuzumab has been introduced into clinical practice for high-risk HER2−positive patients who have completed the standard adjuvant treatment.

However, small (<1cm), node-negative tumors remain a subgroup of HER2−positive patients who are currently ineligible for trastuzumab treatment, as clinically they have been deemed to have no requirement for standard adjuvant chemotherapy.

The aim of our analysis was to evaluate the prognostic factors of local and distant recurrence in patients diagnosed with T1a-b, node-negative, HER-2 positive breast cancer.

Materials and methods. A total of 704 patients were diagnosed at Florence University between November 1999 and December 2008 with node-negative, invasive BC that were 1 cm or smaller. Patients with ductal carcinoma in situ, with recurrent BC at presentation and patients that received adjuvant chemotherapy were excluded from analysis.

Results. Mean follow-up was 4.9 years (0.5−10.8 years); we recorded a total of 19 recurrences, including 10 distant recurrences. Mean time to local relapse occurrence was 3.8 years (0.4−7.3 years); mean time to distant metastases diagnosis was 4.4 years (1.4−7.6 years). Among all patients, the recurrence-free survival (RFS) was 93.7%.

The only parameter that emerged as significant predictor of events is the age (p=0.02).

The distant recurrence-free survival (DRFS) rate was 96.5%. Patients who had HER2−positive BC had worse DRFS than patients who had HER2−negative BC (92.0% v 96.9%; p=0.045). In addition to HER2, also HR status was significantly associated with DRFS (p=0.026). Patients who had HR negative status had worse DRFS than patients who had HR positive status (91.4% v 97.4%; p=0.045).

Conclusions. Patients with T1a-b, node-negative, breast cancer have a low risk of distant and local recurrence. Women with HER2−positive and negative hormonal status have a significant risk of distant recurrence and should be considered for anti-HER2 adjuvant therapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-14.
P2-12-23: How Should We Assess Tumour Size (T Stage) in Patients with Multicentric/Multifocal Breast Cancer? Results from the NCIC CTG MA.5 Randomized Trial of CEF vs. CMF in Pre-Menopausal Women with Node Positive Breast Cancer.
Authors: Bouganim N , Dong B , Hilton JF , Chapman J-AW , Arnaout A , O'Malley F , Nielsen T , Gelmon K , Yerushalmi R , Levine M , Bramwell V , Whelan T , Pritchard KI , Shepherd L , Clemons M
Published: June 2012

Background

A common clinical conundrum in breast cancer management is whether pathologic T stage in women with multicentric or multifocal disease should be taken as the diameter of the largest focus or as the sum of all foci in the breast. Most staging systems, such as the American Joint Committee on Cancer (AJCC), simply use the largest tumour focus for staging. We examine here the impact of alternate methods of estimating tumour size including measures of total tumour size, volume and surface area.

Methods: NCIC CTG MA.5 is a randomized trial of CEF versus CMF in pre-menopausal women with node positive breast cancer.

Median follow up is 10 years. Pathologically reported patient tumour dimensions for up to 3 foci were utilized to examine the effects of tumour size on Breast-Cancer-Free-Interval (BCFI). BCFI is defined as the time from randomization until recurrence: first local invasive or DCIS, regional, distant, contralateral invasive or DCIS. Tumour size was estimated as 1) pathologic T stage as per AJCC criteria; 2) largest dimension of largest tumour focus (cm); 3) sum of largest dimension(s) of tumour foci (cm); 4) sum of surface area(s) of tumour foci (cm2), and 5) sum of volume of tumour foci (cm3). Step-wise forward unstratified Cox regression was used to assess the different effects of tumour size.

Results: This study accrued 710 patients, 37% with T1 tumours, 52% with T2 tumours and 9% with T3 tumours; 61% had 1 to 3 positive lymph nodes. 59% hormone receptor positive. Higher pathologic T stage (p=0.001) and greater surface area (p=0.02) were associated with shorter BCFI, as was lymphovascular invasion (p=0.03), and # of lymph nodes involved (p<0.0001). Administration of anthracycline therapy led to significantly longer BCFI (0.003). The sum of largest tumour sizes (p=0.33) and sum of tumour volume (p=0.34) were not significantly associated with BCFI. Additionally, when the less complete locally reported tumour grade data were included, higher tumour grade was associated with shorter BCFI (p<0.0001).

Conclusions: Consideration of multicentric and multifocal disease was an important adjunct to standard pathologic tumour size as was estimation of tumour surface area in this chemotherapy trial of node positive premenopausal women. However, simply adding together the diameters of tumours in patients with multicentric or multifocal disease did not add any additional prognostic information in this high risk patient population.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-23.
P2-14-08: Hormone Receptor Status and Breast Cancer Survival among Hispanic and Non-Hispanic White Women over 10 Years of Follow-Up.
Authors: Denkhoff SR , Baumgartner KB , Yang D , Pinkston CM , Baumgartner RN
Published: June 2012

Introduction: The joint presence or absence of expression of estrogen receptor (ER+/−), progesterone receptor (PR+/−), and Her2neu (Her2+/−) is now widely used to define the hormone receptor (HR) phenotype of breast tumors. HR phenotype influences breast cancer etiology, response to treatment and survival, which may vary by ethnic group. The triple negative phenotype (TNT: ER-/PR-/Her2-) is associated with the poorest survival and is more common in African American than white women. Few studies have assessed how HR status differs by Hispanic (H) versus non-Hispanic white (NHW) ethnicity in relation to survival.

Methods: We analyzed data from the New Mexico site of the ‘Health, Eating, Activity and Lifestyle’ (HEAL) study for breast cancer mortality over 10 years of follow-up by HR phenotype and ethnicity for 126 H and 408 NHW women with invasive (I-IIIA), first primary breast cancer. Kaplan Meier curves were used to describe differences in survival by ethnicity and HR phenotype (ER+/PR+/Her2+, referent; ER+/PR+/Her2-; ER-/PR-/Her2+; and ER-/PR-/Her2-). Hazard ratios were estimated using Cox regression models stratified by phenotype and ethnicity, adjusting for age at diagnosis, stage, and diagnosis of another cancer (yes/no).

Results: Kaplan Meier (KM) analysis indicated that the TNT phenotype was associated with poorer survival, particularly during the first 5 years, although the difference was not statistically significant after 10 years of follow-up (p=0.36). The prevalence of TNT was higher in H (10.3%) than NHW (6.6%) women, and H also tended to poorer survival than NHW women (p=0.17). In multivariate-adjusted models, women with a TNT phenotype had a 26% increased risk of death (HR=1.26 95% CI 0.62−2.6). In NHW women, the risk was 12% (HR=1.12 95% CI 0.43−2.9). Among H women, those with a TNT phenotype had a 17% increased risk of death (HR=1.17 95% CI 0.35−3.8) while those with ER-/PR-/Her2+ had a 46% increase (HR= 1.46 95% CI 0.40−5.3).

Discussion: Although this analysis is limited by small numbers, the results suggest that H women may have worse 10-year survival rates than NHW women and that women with TNT have a higher risk of death regardless of ethnicity. These differences in survival could be due to a higher rate of recurrence or issues related to treatment. This study suggests that there is an ethnic disparity in survival by HR subtypes that should be examined in a larger population based study.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-14-08.
P2-15-02: Are We Performing More Mastectomies on Women Diagnosed with Invasive Breast Cancer?
Authors: Lo MCI , Hariri B , Gandamihardja T , Lewis J , Hogben K
Published: June 2012

Introduction: Surgical management of breast cancer has changed over the past several decades; breast-conserving surgery has become widely accepted. Mastectomies, however, are still being performed as part of the management of breast cancer. The aim of this study was to investigate the indications for mastectomy in women with invasive breast cancer within our institution.

Method: A systematic review of all new patients with invasive breast cancer treated with surgical intervention at our breast unit in the years 2000, 2005 and 2009 was performed. Using our databases, patient demographics, tumour characteristics and reasons for mastectomy were examined. Screen-detected and symptomatic patients were included in the study. Patients excluded were those with incomplete data and recurrent breast cancer.

Results: A total of 584 patients (F:M, 576:8); with cancer types which included invasive ductal carcinoma (n=501, 84%), invasive lobular carcinoma (n=51, 9%), invasive tubular carcinoma (n=27, 5%) and sarcomas (n=5, 1%) were studied.

Conclusion: In our institution, the majority of patients with screen-detected invasive breast cancer fulfilled the criteria to have breast-conserving surgery; mastectomy rate for screen-detected cancers remained stable. However, we found that in the symptomatic patient group, there was an increase in mastectomy rate from 16% (n=170) in 2000 to 52% (n=225) in 2009. Despite this trend, our results were still in keeping with the national standard as illustrated by the National Cancer Intelligence Network, where they noted that in 2006, 52% of symptomatic patients with invasive breast cancer underwent mastectomy in the United Kingdom.

The reason for a higher mastectomy rate in symptomatic patients compared to screen-detected patients in 2009 is multifactorial. We found that a larger proportion of symptomatic patients presented with larger tumours and tumours which were multifocal in nature. Other factors which may have contributed included more accurate estimation of tumour size by imaging (e.g. MRI) and patient choice.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-15-02.
P2-16-14: Skin Sparing Mastectomy and Immediate Latissimus Dorsi Flap Reconstruction: Patient Reported Outcome and Factors Affecting the Highest Patient Satisfaction.
Authors: Kim Z , Kang S-G , Lee J , Kim SY , Lim CW , Lee MH
Published: June 2012

Background: Skin sparing mastectomy (SSM) and immediate breast reconstruction (IBR) with latissimus dorsi (LD) flap is a tailored surgical procedure for breast cancer patients. In this study the oncologic safety, morbidities, and aesthetic results of SSM and LD IBR, as regards to patient satisfaction, were assessed.

Material and Methods: Between March 2000 and February 2011, single surgeon performed SSM and IBR for 145 patients. Eighty-five patients completed the patient satisfaction survey, and 65 patients with SSM and LD IBR were included. The patients were divided into 2 groups according to their degree of satisfaction, and a stratified analysis was performed.

Results: The mean age of the patients was 48.4 years (range, 21–74), and the pathologic results were infiltrating ductal carcinoma (n=48, 73.8%), ductal carcinoma in situ (n=15, 23.1%), and others (n=2, 3.1%). After a mean follow-up of 34 months (range, 1.6−89.9) no local recurrence occurred, and there were no skin necrosis or LD flap loss. Donor site morbidities were seroma (n=8, 12.3%), scarring (n=8, 12.3%), and back pain (n=6, 9.2%).

Fifty patients (76.9%) were satisfied with their surgical outcomes, and 40% reported their degree of satisfaction as excellent. Breast symmetry (P<0.001), nipple cosmesis (P<0.001), surgeon assessment (P<0.001), observer assessment (P<0.001), and visual difference of bilateral breasts (P=0.021) were factors that affected the highest patient satisfaction.

Conclusion: The present study demonstrated that our SSM and LD IBR was oncologically safe, and was associated with low morbidity rates. LD flap per se, without a prosthetic implant, could sufficiently produce a high level of patient satisfaction. Accurate surgical techniuqes and careful consideration for breast symmetry and nipple cosmesis would be the key to meet the patient's expectations and enhanced quality of life.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-16-14.
P2-18-06: Conventional Trastuzumab Is an Antagonist of Natural Killer Cells: Making the Case for Fucose-Depleted Trastuzumab.
Authors: Listinsky JJ , Siegal GP , Listinsky CM
Published: June 2012

The discovery of the HER2 receptor and later the development of trastuzumab (an anti-HER2 antibody) were major advances leading toward the treatment of a significant proportion of human breast cancers. Trastuzumab (Herceptin®) is a humanized monoclonal IgG1 whose discrete antitumor effects are exerted through the 2 main components of the antibody. Trastuzumab's FAB portion binds to the HER2 extracellular domain, interrupting HER2 signaling. The FC portion then engages the activating receptor (FcγRIIIa) on an effector cell, usually a natural killer (NK) cell and instigates a lytic attack via antibody-dependent cellular cytotoxicity (ADCC).

The value of trastuzumab in breast cancer therapy is well established, but, there are limitations to the drug's effectiveness. These include: (1) unresponsive tumors with low or equivocal HER2 expression; (2) de novo (primary) resistance to therapy in over 50% of patients with HER2−positive tumors; (3) frequent emergence of secondary resistance often after short exposure periods; (4) competition of plasma IgG proteins with antibodies for binding to the activating FC receptors on NK cells; and (5) the high cost of trastuzumab treatment, raising concerns for the economic sustainability of this form of therapy.

Trastuzumab and other therapeutic antibodies share a structural feature that is critical to their effectiveness: a specific oligosaccharide which is covalently linked to the FC region at asparagine 297 (Asn297) in each of the 2 heavy chains. This oligosaccharide, a mannosyl-chitobiose core, is “core-fucosylated” with a single α-L-fucose (fucose) sugar attached via an α(1,6) linkage. In 2002, Shields et al. developed a novel form of trastuzumab which lacked core fucose in the Fc region [J Biol Chem 2002; 277: 26733–40]. When compared to conventional trastuzumab, the fucose-depleted trastuzumab was 43-fold more potent in provoking ADCC against HER2−expressing human breast cancer cells in vitro. Subsequent studies with several monoclonal antibodies have elucidated the role of core fucose in inhibiting ADCC, by reducing the affinity of trastuzumab's Fc portion for the activating FC receptors located on effector cells. The superior ADCC effect of fucose-depleted trastuzumab has now been demonstrated in vivo [Junttila TT et al., Cancer Res 2010; 70: 4481–4489].

We review both in vitro and in vivo evidence from multiple laboratories (working with trastuzumab, rituximab and other therapeutic antibodies) to argue that fucose-depleted trastuzumab offers several potential advantages over conventional trastuzumab. Such advantages in clinical use, we postulate, include (1) treatment of patients with trastuzumab-resistant tumors, (2) treatment of patients with equivocal or low expression of HER2, and (3) decreased cost of treatment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-18-06.
P3-01-07: A Murine Xenograft Model for the Study of Ciclooxygenase-2 Expression and Function in Human Breast Cancer.
Authors: Solano AR , Orlando UD , Garona J , Ripoli GV , Duarte AB , Maloberti PM , Avagnina MA , Alonso DF , Gomez DE , Podesta EJ
Published: June 2012

BACKGROUND: ciclooxygenase-2 (COX-2) has been suggested as a necessary component of the cellular and molecular mechanisms behind breast cancer cell motility and invasion The potential therapeutic benefit of COX-2 inhibitors in a range of cancers is being seen as a great promise however, there have been recent concerns about potential cardiotoxicity. Thus, there is an urgency to develop new inhibitors exhibiting a better risk/benefit ratio.

Acyl-CoA synthetase 4 (ACSL4), belongs to a five-member familyof enzymes that esterifies mainly arachidonic acid into acyl-CoA. We have provided first-time evidence demonstrating that, ACSL4 is the key enzyme that regulates the induction of COX-2, the production of prostaglandin E2 (PGE2) and the proliferation and metastatic potential of breast cancer cells. Therefore, the aim of the study was the development of an in vivo model of human breast tumor xenografts for the study of the regulation of COX-2 expression and action.

RESULTS: stable transfection of MCF-7 cells with ACSL4 cDNA under the control of tetracycline (MCF-7 Tet-off-ACSL4) resulted in a significant increase in the expression of COX-2. The increment in COX-2 expression registered in MCF-7 Tet-off-ACSL4 cells is accompanied with an increase in the production of PGE2, and the proliferation and metastatic potential of cancer cells. Next, we tested whether the injection of MCF-7 Tet-off/ACSL4 cells into nude mice resulted in tumor development.

The results of those experiments demonstrate that MCF-7 Tet-off/ACSL4 cells develop into murine mammary tumors whereas cells transfected with the MCF-7 Tet-off empty vector did not. Interestingly, treatment of nude mice with tetracycline resulted in tumor growth inhibition. The results show that the sole transfection of ACSL4 results in a phenotype change that endows the cells with the capacity to develop into tumors when injected into nude mice. Tumor volume (TV) at day 70 reached 468.4 ± 189.3mm3. Treatment with tetracycline reduced TV to 189.3 ± 65.9mm3 (p<0.05). The growth rate between days 45–70 was 12.30 mm3/day. Tumors were classified as Basal-like subtypes.

We have also observed an inhibition of proliferation and migration of MDA-MB-231 cell cultures when exposed to a combination of ACSL4 and COX-2 inhibitors. Interestingly, the compounds assayed markedly reduced cell proliferation and migration at concentrations that are less effective when used alone. These results point to an effect that has the advantage of exposing the cells to lower drug concentrations.

CONCLUSION: Based on our results, we hypothesize that ACSL4 and COX-2 could constitute potential therapeutic targets for the control of tumor growth. Our animal model of mammary tumors constitutes a proper platform for the study of those therapies.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-01-07.
P3-01-13: Leptin Modulates the Response to Tamoxifen Treatment in ER-Positive Breast Cancer Cell Lines.
Authors: Yom CK , Han W , Kim S-W , Kim DW , Kang E , Noh D-Y
Published: June 2012

Obesity is known to a risk factor for breast cancer in postmenopausal women. Leptin is a potent adipokine which mainly produced by adipose tissue and leptin signaling plays a significant role in tumor development and invasion of breast cancer. In recent study, leptin induced aromatase gene expression, elevating aromatase activity and increasing estrogen synthesis in MCF-7 cell. Leptin was also able to enhance estrogen receptor α (ERα)-dependent transcription by decreasing ERα ubiquitination and degradation. And this imply that lepin may play some role on hormone resistancy of ER-positive breast cancer. The aims of this study is to evaluate whether leptin modulates the effect on tamoxifen treatment in estrogen receptor (ER)-positive breast cancer cell.

At first, we examined leptin and Leptin receptor(Ob-R) expression by western blot assay in various breast cancer cells and cell proliferation by cell viability assay in combination with leptin and tamoxifen under estradiol (E2) treatment. And we analyzed the effect of leptin on the response of tamoxifen treatment in ER-positive breast cancer cells and which result from activation of ERK1/2 and STAT3 signaling pathway in leptin treatment.

We used the MCF-7 (ER+/Leptin +/Ob-R+) cell and T-47D (ER+/ Leptin +/Ob-R-)cell to compare the effect of leptin according to Ob-R status. Leptin had the proliferative effect on MCF-7 comparing to control and T-47D cells. But leptin had no additional effect on both cells in combination with tamoxifen or E2. Interestingly, Leptin had an inhibitory effect on tamoxifen especially in high E2 (20 nM) setting in both MCF-7 and T-47D cells. In cell signaling, when leptin was added to combination treatment of E2 and tamoxifen, the total- and phospho- of ERK1/2 was more activated than combination without leptin in MCF-7. In case of STAT3, activation of total- and phospho-STAT3 were relatively remarkable comparing to combination without leptin, when the three agents are combined to treatment in MCF-7. On the contrary, in T-47D cell, when leptin added to combination of E2 and tamoxifen, leptin relatively induced the activation of total ERK1/2 comparing to combination of E2 and tamoxifen without leptin. But this result was not shown in phospho-ERK1/2 signaling. In STAT3 signaling, same results was noted in T-47D.

We concluded that leptin interfere with the anti-estrogenic effects of tamoxifen under E2 condition both in MCF-7 and in T-47D ER-positive breast cancer cells through activation not only Ob-R but also crosstalk other signaling system. These results imply that inhibition of leptin expect to enhance the hormone response in ER-positive breast cancer, and therefore, to be the promising way to overcome hormonal resistance.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-01-13.
P3-05-06: Progression of Breast Cancer Molecular Subtypes through Different Clinical Stages.
Authors: Ciruelos EM , Castaneda CA , Andrés E , Gomez HL , Castellano D , Mendiola C , Manso L , Ghanem I , Farfan C , Cortes-Funes H
Published: June 2012

Purpose : Molecular classification of breast cancer (BC) through immunohistochemistry classifies patients globally in 4 subtypes with similar features and prognosis. The present study was designed to compare features of BC subtypes in early versus locally advanced clinical stages. A secondary objective was compare molecular subtypes of the primary vs recurrences.

Methods : The study included 1621 patients with non-metastatic invasive BC that were consecutively diagnosed at Hospital Universitario 12 de Octubre, Madrid, between 1997 and 2007. Luminal A was defined as ER+ and PR+, HER2−negative and Ki67 < 14%. Luminal B was defined as PR-negative, HER2−positive or ki67≥14%. HER2 was defined as ER and PR-negative, and HER2−positive.Triple negative (TN) were tumors with ER, PR and HER2 -negativity. Clinicopathological characteristics and outcomes were retrospectively reviewed. Variables were compared with the X2 test, and survival curves were evaluated with Kaplan-Meier method.

Results : Most patients were diagnosed as T1 (48%) and T2 (39.7%) clinical stages, and classified as Luminal B (49%) and Luminal A (29%). GIII frequency increased from T1 to T3-4 in Luminal A (p=0.002), and Luminal B (p=0.051) subtypes, but not in HER2 (p=0.867) or TN (p=0.53).

Molecular subtypes carry significant different prognosis (DFS and OS) in all T stages except for T1a cases. When T3-4 and T1 stages of the same molecular subtype were compared, a significant shorter DFS were found for more advanced stages in the Luminal A (p=0.0002), Luminal B (0.0001) and TN (0.0017), but not for HER2 (p=0.54) subtype. And, similar results were found for OS.

We compared molecular subtypes in the primary tumor and in the metastatic site in 83 cases (excluding contralateral recurrences) and found change of phenotype in 54% of the cases. Changes from Luminal A to a more aggressive phenotype were more frequent than in the opposite situation (14 vs 2 cases).

Conclusion : Despite the molecular classification of early-stage BC that classifies patients in well-defined prognostic subgroups, tumors are continuously changing and tumor behavior becomes more aggressive through progression. Therefore, even tumors with favorable phenotype could loss their good prognosis in locally advanced stages. Obtention of tumor tissue at metastatic sites is also mandatory to a better selection of systemic therapies in relapsed patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-05-06.
P3-06-04: Sno/miRNA Expression Via Next Generation Sequencing: Variation in Patients before and after Treatment.
Authors: Banerjee N , Kamalakaran S , Varadan V , Janevski A , Lezon-Geyda K , Bossuyt V , Flowers D , Sikov W , Abu-Khalaf M , Rizack T , Harris L , Dimitrova N
Published: June 2012

Background

Aberrant expression of small RNA molecules has been shown in breast cancer. It is yet unclear if variation exists in small RNA molecule expression in response to treatment. Since next generation sequencing offers more globally sensitive detection of sno and miRNAs, we performed an RNA-seq study to explore patients pre- and post- brief exposure to treatment.

Methods: We sequenced transcriptomes of frozen biopsy samples from 8 breast cancer patients enrolled in a clinical trial for neoadjuvant therapy using trastuzumab (HER2 positive) or bevacizumab (HER2 negative) with chemotherapy. Tumor core biopsies were taken before and after 10 days of either biologic or nab-paclitaxel treatment and stored in OCT compound. Total RNA was extracted and libraries were constructed for the 16 samples using TruSeq (Illumina). We performed 74bp paired-end sequencing with the Illumina GAII platform. Sequences were aligned to the sno/miRNA track (containing 928 miRNAs and 413 snoRNAs) in UCSC and read counts were determined using Bowtie. We performed differential miRNA and snoRNA expression analysis pair-wise in all pre- and post-therapy samples. Given that miRNA deregulation relies on their protein-coding gene targets, we analyzed the predicted targets of the significantly varying miRNAs for functional enrichment.

Results: Each sample had on average 46 million paired-end reads, of which on average 70% were mapped to the human genome. Overall, we detected 138 miRNAs in at least one sample, with each sample expressing 33 miRNAs on average. We detected a total of 11 miRNAs (7%) that showed significant differential expression with treatment. Interestingly, 6 of these miRNAs varied in all patients. The predicted targets of these miRNAs were enriched in DNA-dependent transcription, gene expression, cell proliferation and cell communication. Similarly, we detected 202 snoRNAs in at least one sample, with each sample expressing 87 snoRNAs on average. Of these, we found 21 snoRNAs (10%) to vary significantly upon treatment and 6 of these snoRNAs showed expression changes in all patients.

Conclusions: These results suggest that variation in sno/miRNA expression may play a role in response to treatment. The consistent variation of sno/miRNAs in response to treatment suggests shared small RNA-mediated mechanisms. If validated, these results suggest that next generation sequencing technologies will allow lead to improved methods of stratifying, subclassifying and managing breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-06-04.
P3-07-05: Nomogram Incorporating SLN Metastasis Size Provides the Most Accurate Prediction of Non-SLN Involvement in Breast Cancer Patients.
Authors: Gainer SM , Mittendorf EA , Hunt KK , Bassett RL , Yi M , Meric-Bernstam F , Ross MI , Babiera GV , Kuerer HM , Caudle AS , Hwang RF
Published: June 2012

Introduction: The finding of metastatic disease in the axillary nodes remains an important factor that guides adjuvant therapy decisions in breast cancer. The value of completion axillary lymph node dissection (ALND) may not be the same in all patients and determining the risk of non-SLN involvement can be a guide in surgical decision making. We recently developed a nomogram that incorporates the maximum SLN metastasis size to predict non-SLN axillary involvement (Mittendorf et al., Ann Surgery 2011,in press). The purpose of this study was to compare the accuracy of this nomogram to another widely used nomogram that does not include SLN metastasis size.

Methods: We queried a prospective breast cancer database for patients with clinically negative nodes undergoing sentinel node biopsy who did not receive neoadjuvant therapy. We identified 431 patients with invasive breast cancer treated from 1996 to 2007 who had a positive SLN and underwent completion ALND. We evaluated clinicopathologic data including histology, multifocality, nuclear grade, pathologic tumor size, number of SLNs recovered, number of positive SLNs, maximum SLN metastasis size, presence of extranodal extension, hormone receptor status, and presence of lymphovascular invasion. Data were entered into our nomogram and the nomogram that does not include SLN metastasis size. The accuracy of the 2 nomograms in predicting non-SLN axillary metastasis was compared in all patients and in subgroups of patients based on burden of disease in the SLN.

Results: For the entire cohort of 431 women with positive SLNs, our SLN size nomogram was more accurate in the detection of non-SLN disease (area under the receiver operating characteristic curve [AUC] 78.3% vs. 72.4%, p=0.01). Whether or not SLNs were subjected to frozen section did not influence the relative accuracies of the 2 nomograms. When patients were analyzed by burden of disease in the SLNs, our SLN size nomogram was more accurate in predicting non-SLN disease in patients with macrometastases in the SLN (AUC 77.3% vs. 70.3%, p=0.008). There was no significant difference in the accuracy of the 2 nomograms for patients with isolated tumor cells or micrometastases in the SLNs.

Conclusions: Our nomogram incorporating SLN metastasis size with other clinicopathologic factors is the most accurate predictor of the risk of non-SLN metastasis in patients with a positive SLN. The SLN size nomogram is highly accurate in women with macrometastases, who may benefit the most from completion ALND. Using a nomogram with SLN metastasis size can provide the most accurate information for patient counseling and surgical decision making.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-05.
P3-07-06: Prognostic Utility of Upfront Nodal Staging Prior to Neoadjuvant Chemotherapy: The UAB Experience.
Authors: Keene KS , De Los Santos JF , Krontiras H , Hinton B , Meredith R , Li Y , Carpenter JT , Bland K , Forero A
Published: June 2012

Purpose: Controversy exists regarding the prognostic utility of upfront lymph node staging in patients receiving neoadjuvant chemotherapy. This retrospective study explores whether upfront verses outback nodal staging influenced locoregional control and survival endpoints. Methods: Between 1999 and 2005 one hundred and fifteen patients treated with neoadjuvant chemotherapy at UAB were identified. Patient, tumor, and treatment variables were recorded. Timing of nodal assessment was based on either surgeon preference or stipulated upfront per several clinical protocols. Survival was measured using the Kaplan Meier statistics. Univariate and multivariate analyses of covariates associated with local-regional control (LRC), progressionfree (PFS) and overall survival (OS) were performed. Results: Mean age was 49 years and mean follow-up was 5.8 years. Stage distribution was as follows: 40 IIA, 34 IIB, 26 IIIA, 10 IIIB, and 5 IIIC. Definitive surgery included breast conservation in 49 patients, total mastectomy in 21 and modified radical mastectomy in 44. Seventy-two patients had upfront nodal sampling before neoadjuvant therapy, 36 by fine needle assessment and the remainder by sentinel node biopsy. Forty-three patients had their nodal assessment following neoadjuvant chemotherapy. Of those with upfront nodal staging: forty nine patients had a positive nodal result and 23 had negative findings. In those that were sampled at the time of definitive surgery: 21 had positive results, 21 had a negative result, and 1 patient did not have any nodes in the specimen. One hundred five patients had post-operative radiation therapy. Overall there was no difference in LRC, PFS or OS outcomes between patients that had an upfront nodal staging procedure and those that had their nodes sampled at the time of definitive surgery. Patients achieving pCR had a non-significant trend towards improved overall survival (p=0.12). Final pathologic T, N and group stage were statistically significant in determining the progression free, p=0.003, 0.011, 0.005 and overall survival, p=0.02, 0.037, and 0.009. Lymphovascular space invasion was associated with progression free survival on univariate analysis, p=0.0179, but not multivariate analysis. Other covariates including age, biologic subtype, grade, type of chemotherapy, use of radiation therapy, radiation volume, time to complete radiation therapy, and use of hormonal therapy did not affect outcomes. Discussion: Upfront as compared with outback lymph node staging did not influence locoregional control or survival endpoints. Final pathologic stage and not initial clinical stage remains the most important prognostic factor associated with survival.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-06.
P3-07-12: Value of Preoperative Ultrasound-Guided Axillary Lymph Node Biopsy for Preventing Futile Sentinel Node Procedures in Breast Cancer: A Systematic Review and Meta-Analysis.
Authors: Diepstraten SC , Sever AR , Buckens CF , Veldhuis WB , van den Bosch MA , Mali WP , Verkooijen HM
Published: June 2012

Purpose: Women affected with invasive breast cancer generally undergo sentinel node biopsy (SNB) of the axilla, which is followed by axillary dissection if lymph node metastases are found. This systematic review and meta-analysis aims to evaluate the utility of preoperative axillary ultrasound-guided needle biopsy in terms of staging the axilla and preventing futile SNB.

Methods: We systemically searched Pubmed, Embase and Cochrane through March 14, 2011, for studies addressing preoperative assessment of axillary lymph node (ALN) status by axillary ultrasound and biopsy. Eligible studies had to report on biopsy indication and method, and true and false positive and negative outcomes. A pooled estimate was calculated for prevalence of futile SNB (defined as the proportion of patients needing secondary ALN dissection after a negative axillary ultrasound and biopsy) and sensitivity (defined as the proportion of patients with ALN involvement detected by axillary ultrasound and biopsy).

Results: Twenty-three studies were included which reported on a total of 6205 procedures. The pooled prevalence of futile SNB was 25% (95% CI = 23% to 27%) and the pooled sensitivity was 51% (95% CI = 43% to 59%). There was substantial heterogeneity across studies for both futile SNB prevalence (I2 = 60.5) and sensitivity (I2 = 93.3), which could not be explained by between-study differences in biopsy technique, inclusion criteria, biopsy indication or study design. Sensitivity was increased in studies with a high prevalence of axillary lymph node metastases.

Conclusion: Preoperative axillary ultrasound guided biopsy is a useful step in the process of axillary staging as around fifty percent of breast cancer patients with axillary involvement can be identified pre-operatively and are spared a two step surgical intervention. Still, one in four women with a US biopsy ‘proven’ negative axilla have ALN involvement on SNB and need to undergo completion axillary dissection. New techniques may be able to reduce the prevalence of futile SNB.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-12.
P3-07-17: Optimal Number of Sentinel Lymph Nodes That Could Avoid Completion of Axillary Lymph Node Dissection in Operable Breast Cancer.
Authors: Salmon RJ , de Ricke Y , Falcou MC
Published: June 2012

Background

Axillary lymph node dissection (ALND) has been the standard of care for breast cancer patients with sentinel lymph node (SLN) metastasis. It is now under discussion since the publications of ACOZOG Z0011 and NSABP-32. The 1–3% rate of axillary recurrence is very low in these studies, whereas the rate of additional nodal metastasis after the completion of ALND varies from 15% to 35%. The purpose of this study was to determine the optimal number of nodes to be removed in order to obtain an axilla free of disease after initial surgery.

Patients and methods: 4928 consecutive patients treated for breast cancer ≤ 2 cm with primary breast surgery with SNB between 2000 and 2009 were reviewed from the Institut Curie breast cancer prospective database. 1165 patients with a positive SLN (macrometastasis, micrometastasis and isolated tumor cells) underwent ALND. The proportion of patients with positive ALND was analysed according to the total number of sentinel lymph nodes biopsied. Results were compared with Chi-square test for qualitative variables and with Kruskal-Wallis for quantitative variables.

Results: Among the 1165 patients, 308 (26.4%) had a positive ALND. Among them, 81.1% of patients had a macrometastasis in the SLN versus 45.6% in the 857 patients with negative ALND (p<0.0001); 10.8% had a micrometastasis versus 31.4% (p<0.001) and 8.1% of patients had isolated tumor cells in the SLN versus 23% (p<0.001). The proportion of patients with positive ALND decreased with the number of sentinel nodes removed (Figure 1), independently of the number of positive SLN. Among the 1165 patients, the rate of patients with 5 nodes removed or more and with residual disease in the axilla was < 1% (n=11). Those patients could have avoided completion ALND, representing 3.6% of the total positive ALND that would have been neglected.

Conclusions: We showed that patients with positive SLN have a very low risk (<1%) of residual disease in the axilla when 5 nodes or more have been removed. This number which is a major component of nomograms, could be used as a threshold to avoid completion ALND.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-07-17.