Factor In Triple-negative Breast Cancer Research Articles

Decreased Wnt5a Expression is a Poor Prognostic Factor in Triple-Negative Breast Cancer.

BackgroundWingless-type MMTV integration site family member 5A (Wnt5a) has been documented to either overexpress or be lost in several malignancies. Our study aimed to investigate the expression and clinical significance of Wnt5a protein in triple-negative breast cancer (TNBC).Material/MethodsBy using immunohistochemistry, Wnt5a expression was evaluated in 90 TNBC specimens. The association between Wnt5a expression and clinic-pathological factors was assessed by using the chi-square test. The survival analysis of patients was conducted by using the Kaplan-Meier and log-rank tests. Cox regression was utilized for the univariate and multivariate analyses of prognostic factors.ResultsResults showed that negative Wnt5a expression correlated with positive lymph node metastasis (LNM) (P=0.007) and Ki67 proliferation (P=0.002). Patients with negative Wnt5a expression had significantly poorer recurrence-free survival (RFS) than patients with positive Wnt5a expression (P=0.008). Multivariate Cox regression analysis revealed that decreased Wnt5a expression was an independent prognostic factor for RFS (P=0.014).ConclusionsNegative Wnt5a protein expression might contribute to the tumor progression and poor prognosis of TNBC and might be a new therapy target in TNBC.

Myeloid cell leukemia-1 is an important apoptotic survival factor in triple-negative breast cancer

Breast cancer is the second-most frequently diagnosed malignancy in US women. The triple-negative breast cancer (TNBC) subtype, which lacks expression of the estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2, afflicts 15% of patients and is refractory to current targeted therapies. Like many cancers, TNBC cells often deregulate programmed cell death by upregulating anti-apoptotic proteins of the B-cell CLL/lymphoma 2 (Bcl-2) family. One family member, myeloid cell leukemia-1 (Mcl-1), is commonly amplified in TNBC and correlates with a poor clinical prognosis. Here we show the effect of silencing Mcl-1 and Bcl-2-like protein 1 isoform 1 (Bcl-xL) expression on viability in a panel of seventeen TNBC cell lines. Cell death was observed in a subset upon Mcl-1 knockdown. In contrast, Bcl-xL knockdown only modestly reduced viability, indicating that Mcl-1 is a more important survival factor. However, dual silencing of both Mcl-1 and Bcl-xL reduced viability in most cell lines tested. These proliferation results were recapitulated by BH3 profiling experiments. Treatment with a Bcl-xL and Bcl-2 peptide had only a moderate effect on any of the TNBC cell lines, however, co-dosing an Mcl-1-selective peptide with a peptide that inhibits Bcl-xL and Bcl-2 was effective in each line tested. Similarly, the selective Bcl-xL inhibitor WEHI-539 was only weakly cytotoxic across the panel, but sensitization by Mcl-1 knockdown markedly improved its EC50. ABT-199, which selectively inhibits Bcl-2, did not synergize with Mcl-1 knockdown, indicating the relatively low importance of Bcl-2 in these lines. Mcl-1 sensitivity is not predicted by mRNA or protein levels of a single Bcl-2 family member, except for only a weak correlation for Bak and Bax protein expression. However, a more comprehensive index composed of Mcl-1, Bcl-xL, Bim, Bak and Noxa protein or mRNA expression correlates well with Mcl-1 sensitivity in TNBC and can also predict Mcl-1 dependency in non-small cell lung cancer cell lines.

S100A14 is a novel independent prognostic biomarker in the triple-negative breast cancer subtype.

Triple-negative breast cancer (TNBC) represents a heterogeneous subgroup with generally poor outcome and lack of an effective targeted therapy. Prognostic or predictive biomarkers to guide treatment decisions for this group of patients are needed. To evaluate the potential of S100A14 protein as a novel biomarker in TNBC, the protein expression of S100A14 was correlated with clinical outcomes in a Pilot Sample set and a Danish cohort of predominantly TNBC patients. Kaplan-Meier analysis identified a prognostic impact of S100A14 on disease-free survival and overall survival, showing that tumors with high S100A14 protein expression levels were significantly correlated with poor outcome in TNBC patients (p = 0.017; p = 0.038), particularly those in the basal-like subgroup (p = 0.006; p = 0.037). Importantly, TNBC patients with high S100A14 expression, but tumor-negative axillary lymph nodes (N-), had equally poor outcomes as those with tumor-positive axillary lymph nodes (N+), while TNBC/N- patients with low S100A14 expression had a significantly better disease free survival (p = 0.013). Multivariate analysis revealed that S100A14 is an independent prognostic factor for TNBC patients (p = 0.024; p = 0.05). At the cellular level, S100A14 was found to be expressed in epithelial-like, but not in mesenchymal-like, TNBC cells in vitro. S100A14 is an independent prognostic factor in TNBC and a novel potential therapeutic target in TNBC.

Correlation between (18)F-FDG uptake on PET/CT and prognostic factors in triple-negative breast cancer.

The purpose of this study was to investigate whether a correlation exists between (18)F-fluorodeoxyglucose (FDG) uptake and prognostic factors in triple-negative breast cancer (TNBC). Between January 2009 and December 2013, 103 patients (mean age, 50.6 years) with primary TNBC (mean, 2.6 cm; range, 1.0-6.5 cm) underwent (18)F-FDG PET/CT for initial staging. Correlations between maximum standardized uptake value (SUVmax) on PET/CT and prognostic factors including tumour size, nodal status, histological grade, Ki-67 proliferation index, tumour suppressor p53, and 'basal-like' markers (epidermal growth factor receptor and CK 5/6) were assessed. The mean SUVmax of the 103 tumours was 10.94 ± 5.25 (range: 2-32.8). There was a positive correlation between SUVmax and Ki-67 (Spearman's rho = 0.29, P = 0.003) and tumour size (Spearman's rho = 0.27, P = 0.006), whereas this relationship was not observed in the nodal status, histological grade, p53 status and 'basal-like' phenotypes. In a multivariate regression analysis, Ki-67 (P < 0.001) and tumour size (P = 0.009) were significantly associated with SUVmax in TNBCs. Increased (18)F-FDG uptake on PET/CT was correlated with a high Ki-67 proliferation index and larger tumour size in TNBC. These results suggest a potential role of (18)F-FDG PET/CT in identifying TNBC with more aggressive behaviour. • A wide range of FDG uptake reflected heterogeneity of cancer metabolism. • FDG uptake was correlated with the Ki-67 proliferation index in TNBC. • FDG uptake was correlated with tumour size in TNBC. • FDG uptake was not correlated with 'basal-like' phenotype.

Expression of FGFR1 is an independent prognostic factor in triple-negative breast cancer.

Triple-negative breast cancers (TNBCs) are clinically aggressive tumors with limited treatment options. We examined the clinicopathological associations and prognostic implications of FGFR1 and FGFR2 expression in TNBCs. Tissue microarrays constructed from TNBCs were immunostained with FGFR1 and FGFR2, and scored by intensity and percentage of tumor cells stained per intensity for each subcellular compartment, which were correlated with clinicopathological parameters and survival. Cell migration following siRNA-mediated silencing of the FGFR1 gene in TNBC cell lines was also performed. 714 cases were informative for FGFR1 and FGFR2 immunostaining. Thresholds were defined as at least 1% of cells stained and H-score of 100 or more. Proportions positive by each threshold were, respectively, 89.9%, 7.1% for FGFR1 (cytoplasm); 36.8%, 7.8% for FGFR2 (cytoplasm); and 33.5%, 5.2% for FGFR2 (membrane). Significant associations included FGFR1 and FGFR2 immunostaining for lobular subtype, FGFR2 immunostaining with lower grade, and more basal-like cancers with H-scores of 100 or more FGFR1 immunostaining. Multivariate Cox regression analysis showed FGFR1 expression in TNBCs to be independently prognostic for overall survival (OS) at both thresholds. Cases completely negative (less than 1% staining) for FGFR1 immunostaining showed improved OS, while those with H-score of 100 or more immunostaining had the worst OS. Cell line studies revealed up-regulation of the FGFR1 gene in the MDA-MB-231 and Hs578T TNBC cells, and specific knockdown of FGFR1 expression significantly reduced cell migration in MDA-MB-231 cell line. In conclusion, FGFR1 expression in TNBCs is independently prognostic of OS, and H-score of 100 or more FGFR1 immunostaining may define tumors that have treatment potential via FGFR signaling inhibition.

Biologic roles of estrogen receptor-β and insulin-like growth factor-2 in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) occurs in 10–15% of patients yet accounts for almost half of all breast cancer deaths. TNBCs lack expression of estrogen and progesterone receptors and HER-2 overexpression and cannot be treated with current targeted therapies. TNBCs often occur in African American and younger women. Although initially responsive to some chemotherapies, TNBCs tend to relapse and metastasize. Thus, it is critical to find new therapeutic targets. A second ER gene product, termed ERβ, in the absence of ERα may be such a target. Using human TNBC specimens with known clinical outcomes to assess ERβ expression, we find that ERβ1 associates with significantly worse 5-year overall survival. Further, a panel of TNBC cell lines exhibit significant levels of ERβ protein. To assess ERβ effects on proliferation, ERβ expression in TNBC cells was silenced using shRNA, resulting in a significant reduction in TNBC proliferation. ERβ-specific antagonists similarly suppressed TNBC growth. Growth-stimulating effects of ERβ may be due in part to downstream actions that promote VEGF, amphiregulin, and Wnt-10b secretion, other factors associated with tumor promotion. In vivo, insulin-like growth factor-2 (IGF-2), along with ERβ1, is significantly expressed in TNBC and stimulates high ERβ mRNA in TNBC cells. This work may help elucidate the interplay of metabolic and growth factors in TNBC.

Prognostic value of tumor-infiltrating lymphocytes in triple-negative breast cancers from two phase III randomized adjuvant breast cancer trials: ECOG 2197 and ECOG 1199.

Recent studies suggest that tumor-infiltrating lymphocytes (TILs) are associated with disease-free (DFS) and overall survival (OS) in operable triple-negative breast cancer (TNBC). We seek to validate the prognostic impact of TILs in primary TNBCs in two adjuvant phase III trials conducted by the Eastern Cooperative Oncology Group (ECOG). Full-face hematoxylin and eosin–stained sections of 506 tumors from ECOG trials E2197 and E1199 were evaluated for density of TILs in intraepithelial (iTILs) and stromal compartments (sTILs). Patient cases of TNBC from E2197 and E1199 were randomly selected based on availability of sections. For the primary end point of DFS, association with TIL scores was determined by fitting proportional hazards models stratified on study. Secondary end points were OS and distant recurrence–free interval (DRFI). Reporting recommendations for tumor marker prognostic studies criteria were followed, and all analyses were prespecified. The majority of 481 evaluable cancers had TILs (sTILs, 80%; iTILs, 15%). With a median follow-up of 10.6 years, higher sTIL scores were associated with better prognosis; for every 10% increase in sTILs, a 14% reduction of risk of recurrence or death (P = .02), 18% reduction of risk of distant recurrence (P = .04), and 19% reduction of risk of death (P = .01) were observed. Multivariable analysis confirmed sTILs to be an independent prognostic marker of DFS, DRFI, and OS. In two national randomized clinical trials using contemporary adjuvant chemotherapy, we confirm that stromal lymphocytic infiltration constitutes a robust prognostic factor in TNBCs. Studies assessing outcomes and therapeutic efficacies should consider stratification for this parameter.

Molecular breast cancer subtypes and therapies in a public hospital of northeastern Brazil.

BackgroundThe frequencies of molecular breast cancer subtypes vary among different human populations. The Northeastern region of Brazil has a mixed population of African, Indigenous and European ancestry. This retrospective study investigated breast cancer subtypes and applied therapies in a public hospital of Northeastern Brazil.MethodsData of 633 patients with invasive breast cancer from 2005 to 2011 were obtained from medical records. Status of hormone receptor (HR), HER2 and Ki67 expression index of 269 out of 633 patients were used to define subtypes of Luminal A and B, HER2 and triple negative (TN) breast cancer. Expression index of Ki67 ≥ 14% was applied to distinguish Luminal A from Luminal B subtypes.ResultsOverall, 185 (68.77%) and 132 (49.07%) patients showed positive hormone receptor (HR+) and positive HER2 (HER2+) tumors. The mean age ranged from 53.33 to 58.25 years for patients with tumors of Luminal B and Luminal A subtypes, respectively (p = 0.0182). In general, 67.39% of patients with TN tumors aged over 50 and 19.57% aged between 31 and 40 years (p = 0.0046). The rate of small tumors (T1: ≤ 2.0 cm) varied from 22.73% to 52.46% for TN and Luminal A subtypes (p = 0.0088). The rate of high graded (G3) tumors was increased for HER2 and TN subtypes (35.29% and 34.28%) compared to Luminal A and Luminal B subtypes (3.92% and 12.62%), respectively (p < 0.0001). The five-year survival rate ranged from 92.86% to 75.00%, for Luminal A, HER2 and TN subtypes, respectively (HR: 0.260 to 1.015; 95% CI: 0.043 to 3.594; p = 0.2589). Patients with HER2 positive (HER2+) breast tumors did not receive immunotherapy and chemotherapy application varied from 54.84% to 86.49% for Luminal A and HER2 subtypes, respectively (p = 0.0131).ConclusionsThe results of this study revealed a high percentage of HER2+ breast tumors and an increased rate of patients with TN tumors aged over 50 years. This emphasizes the need for establishing immunotherapy as an additional therapeutic option to improve clinical outcomes for patients with HER2+ tumors and to investigate the risk factors of TN breast cancer.

Patterns and risk factors of recurrence in triple-negative breast cancer

To analyze the recurrence characteristics of triple-negative breast cancer (TNBC) and explore their clinicopathological correlations in northern China. Regression analyses were performed for 4 579 breast cancer patients from September 2002 to September 2007. Among them, there were 787 TNBC patients. The median follow-up period was 72 (9-116) months. And 187 (23.8%) TNBC patients recurred and it was significantly different from the recurrence rate of non-TNBC group (17.4%, P = 0.024). The median time to recurrence was 30.2 (4-110) months and 36.3% recurred 2 years while 27.8% did within 2-3 years. The rates of lung, liver and brain metastases to other sites were higher in TNBC patients than those in non-TNBC patients. TNBC patients had higher local recurrence rates than non-TNBC counterparts (2.92% vs 1.56%) (P = 0.009). Age, regional lymph node infiltration, tumor size and pathological stage were all independent prognostic factors of TNBC. TNBC has unique recurrence characteristics. It is prone to recur shortly with a high local recurrence rate. Metastatic rate is higher in visceral organ than bone.

Prognostic and predictive value of tumor infiltrating lymphocytes (TIL) in two phase III randomized adjuvant breast cancer (BC) trials.

11087 Background: TIL have been proposed as a new prognostic factor for triple negative BC [Dieci MV, 2014]. Here we seek to evaluate the prognostic and predictive role of TIL in BC patients enroll...

Pretreatment platelets count levels as a prognostic factor in triple-negative breast cancer in a Hispanic cohort.

1115 Background: Platelet count has been reported to have predictive value in various cancer entities. Several studies suggest a negative impact of thrombocytosis for patient's survival in different types of cancer. In the case of breast cancer, evidence about involvement of platelets is still inconclusive. Therefore, the aim of our study was to assess the influence of pretreatment platelet count levels on survival and establish its prognostic relevance for breast cancer patients in a large cohort of triple negative breast cancer patients. Methods: We retrospectively studied patients diagnosed with primary triple negative breast cancer that had completed all phases of primary treatment from 2005 to 2012, finding 118 patients with platelet counts prior to therapy and clinical follow-up. Baseline platelet numbers were evaluated at diagnosis (before the start of any therapy for disease). Platelet levels were categorized into the following three groups: normal low (< 200 g/L), normal high (200-400 g/L) and thrombocytosis (> 400 g/L). Overall survival (OS) was assessed using the Kaplan-Meier method and were compared using the LogRank test. To evaluate the independent prognostic significance of NLR, multivariate Cox regression models were used. Results: Patients with lower platelets count (< 200 g/L) showed significantly lower median overall survival than those with normal high (200-300 g/L) and thrombocytosis (> 400 g/L) (33 months vs. 71 months, log Rank p=0.024). Pretreatment thrombocytosis was observed only in 9 patients (7.6%). Low platelets counts along with advanced stage were independently correlated with poor prognosis, with hazard ratio 1.2 (p 0.02), and 2.1 (p 0.001), respectively. Estimated media OS, for patients with low platelets count, normal high count and thrombocytosis were 56, 38 and 36 months respectively. Conclusions: In our retrospective study, normal low platelet counts at time of diagnosis were associated with poor prognosis in triple negative breast cancer. We hypothesize that platelets in normal high levels as scavenger of VEGF and therefore as a potent antiangiogenic cellular component of the tumor microvasculature in triple negative breast cancer can also be considered.

Associations between mammography and ultrasound imaging features and molecular characteristics of triple-negative breast cancer.

The triple-negative breast cancer (TNBC) is an aggressive cancer characterized by the absence of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). Preoperative mammography and ultrasound features of TNBC may potentially suggest characteristics of the disease and assist in treatment decisions. The study covered 153 patients with TNBC from May 2011 to May 2012 who were confirmed by postoperative pathology results in our hospital. We compared the radiological findings among the patients and sought to determine the significant iconographic features. The biomarkers p53 and Ki-67 are regarded as significant factors in TNBC. They were therefore used to divide the TNBC into four groups for assessment of relationships with TNBC imaging features. On mammography, most TNBCs exhibit obscure (44.3%) masses. On ultrasound, the majority of masses (95.4%) were predominantly indistinct (50.7%), irregular (76.0%) or featuring posterior echo enhancement/shadowing. Color Doppler flow imaging (CDFI) emphasized hypervascular (32.9%) masses. Differences in CDFI by ultrasound among the four groups were statistically significant (p=0.009). There were obvious differences in the percentages of spiculated margin (p=0.049) and intensive posterior echo (p=0.006) with spotty flow imaging by ultrasound between the Ki-67 (+) p53 (+) and other groups. A combination of mammography and ultrasound revealed the imaging characteristics of TNBC included an obscure mass with less attenuated posterior echoes and some vascularity. A worse prognosis was associated with spiculated margin and intensive posterior echoes with spotty flow imaging.

Association between single nucleotide polymorphisms (SNPs) of XRCC2 and XRCC3 homologous recombination repair genes and triple-negative breast cancer in Polish women.

XRCC2 and XRCC3 genes involved in homologous recombination repair (HRR) of DNA and in the maintenance of the genome integrity play a crucial role in protecting against mutations that lead to cancer. The aim of the present work was to evaluate associations between the risk of triple-negative breast cancer (TNBC) and polymorphisms in the genes, encoding for two key proteins of HRR: XRCC2 Arg188His (c. 563 G>A; rs3218536, Genbank Accession Number NT 007914) and XRCC3 Thr241Met (c. 722 C>T; rs861539, Genbank Accession Number NT 026437). The polymorphisms of the XRCC2 and XRCC3 were investigated by PCR–RFLP in 70 patients with TNBC and 70 age- and sex-matched non-cancer controls. In the present work, a relationship was identified between XRCC2 Arg188His polymorphism and the incidence of triple-negative breast cancer. The 188His allele and 188His/His homozygous variant increased cancer risk. An association was confirmed between XRCC2 Arg188His and XRCC3 Thr241Met polymorphisms and TNBC progression, assessed by the degree of lymph node metastases and histological grades. In conclusion, XRCC2 Arg188His and XRCC3 Thr241Met polymorphisms may be regarded as predictive factors of triple-negative breast cancer in female population.

Abstract P3-14-19: Panitumumab in combination with FEC 100 (5-fluorouracil, epirubicin, cyclophosphamide) followed by docetaxel for operable, triple negative breast cancer (TNBC): Patient outcome

Abstract Background: Panitumumab is an antibody targeting the epidermal growth factor receptor (EGFR) for which an important role has been suggested in TNBC. Consequently, we evaluated a combination of the standard chemotherapy (FEC 100 followed by docetaxel) with panitumumab as neoadjuvant therapy of operable TNBC. Complete pathologic response (pCR) was the primary endpoint, with clinical response, toxicity, and outcome as secondary endpoints. An investigation of biomarkers possibly predictive of pCR accompanied this trial. Here we focus on tumor recurrence, after a median follow up of 33 months [25-40] as on April, 1, 2013. Methods: Sixty patients (pts) with stage II-IIIA TNBC were prospectively included. Systemic neoadjuvant treatment (ST) consisted of the anti-EGFR antibody panitumumab combined with FEC 100, followed by 4 cycles of docetaxel. All pts underwent surgery after ST completion. Patient characteristics: median tumor size: 40 mm [20-120]; SBR grade III: 71.7%; pCR rate: 55.3% and 46.8% (the Sataloff and the Chevallier classifications, respectively). Paraffin-embedded and frozen tumor samples were collected before and after ST for biomarker analysis. EGFR, IGF-1R, MET, cytokeratins 5/6 and 8/18, PTEN, P-cadherin, ALDH1, Ki-67, p53, tumoral FOXP3 expression and the number of FOXP3+ or CD8+ tumor-infiltrating lymphocytes (TIL) were evaluated by immunohistochemistry. Results :.We have observed 9 recurrences: 1 local and 8 distant recidives, including 1 both local and distant. The distant recidives (metastases) were as follows: brain (4 pts); brain and lungs (1 pt); lungs only (1 pt), pleura (1 pt); liver (1 pt). 6 out of the 8 metastatic pts died and all were non-pCR post-ST. The 2 alive pts had brain metastases, but reached a pCR after the ST. Among the 9 relapsed pts 6 were 55 years old or less at the diagnosis. Seven out of those 9 pts had tumors with the clinical size equal or higher than 4 cm. As previously reported (SABCS 2012, abstract 1081), the pCR-predictive biomarkers in this study were high CD8+ TIL count (p = 3.4.10−6) and high ratio between the CD8+ and FOXP3+ TIL counts (CD8+/FOXP3+ &amp;gt; 1.23, p = 8.5.10−5). With this in mind, we have evaluated whether those parameters, assessed before or after the ST, could predict the recurrences. No difference was found in the preoperative CD8+ and the FOXP3+ TIL counts, as well as in the CD8+/FOXP3+ ratio, between the patiens who have recurred and the others. Conclusion : As it has been reported in previous studies, in our cohort of TNBC pts, the relapses occurred early after the administration of the last systemic treatment. The patients who relapsed died rapidly and most of them have not reached pCR after the ST. In addition, half of the metastatic pts got brain deposits. This implies that research on the resistance factors in TNBC should focus on those important for seeding of the “sanctuaries”, like brain. This research is ongoing in our group. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-14-19.

TMPRSS4 as a Poor Prognostic Factor for Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) is characterized by the lack of immunohistochemical staining for estrogen receptors (ER), progesterone receptors (PR), and lack of overexpression or amplification of human epidermal growth factor receptor 2 (HER2). Our aim was to investigate the expression of transmembrane protease, serine 4 (TMPRSS4) in TNBC patients and its possible relationship to the outcome of the disease. A total of 72 TNBC patients and 109 non-TNBC patients who were diagnosed between 2003 and 2008 were enrolled in this study. Immunohistochemistry was used to compare the expression pattern of TMPRSS4 in TNBC and non-TNBC groups, and the prognostic significance was assessed by Kaplan-Meier analysis and Cox proportional hazards regression in TNBC patients. The rate of high expression of TMPRSS4 was significantly higher in TNBC group than that in non-TNBC group. High expression of TMPRSS4 was significantly correlated with lymph node metastasis, histological grade, and tumor size. TNBC patients with high TMPRSS4 expression showed the poorer overall survival (OS) and disease-free survival (DFS) than those patients with low TMPRSS4 expression. In multivariate analysis, only lymph node metastasis and TMPRSS4 expression were the independent prognostic factors for OS and DFS in TNBC. Our study provides evidence that TMPRSS4 expression is associated with lymph node metastasis, tumor size, and histological grade in TNBC patients, and also is an independent prognostic factor for TNBC.

Insurance status as a strong predictor of outcome in triple-negative breast cancer (TNBC): A multi-institutional retrospective study.

1069 Background: TNBC accounts for about 15-20% of all breast cancer. TNBC has particularly aggressive clinical course and accounts for a disproportionate number of breast cancer relapses and deaths in early stage disease. TNBC also have worse prognosis especially in African American (AA) women compared with white women. This retrospective study aims to investigate various clinical and demographic prognostic factors in TNBC. Methods: 476 TNBC patients who were treated at Eastern Carolina University and University of North Carolina, CH between 4/96 and 9/11 were included in this analysis. We collected data on age, race, grade, lymphovascular invasion (LVI), stage, treatments including surgery, chemotherapy, radiation, chemotherapy drugs, insurance type and year of treatment. Overall Survival (OS) was computed from the date of diagnosis to the date of death or last FU. For disease free survival (DFS), patients were scored if they failed either locally or distally. The Cox proportional-hazards regression model was used to compute hazard ratios. Results: The median age was 52 years (21-88 years) with median FU of 3.7 years. 49% women were white race followed by AA 223 (47%) and Hispanic or other race (5%). Stage (p&lt;0.001), grade (p=0.02), surgery (p&lt;0.0001), adjuvant chemotherapy (p=0.025), LVI (p=0.05) and type of insurance were significant predictor of OS. Similarly, stage (p&lt;0.0001), surgery (p&lt;0.0001), LVI (p=0.03) and insurance were significant predictors for DFS. On multivariate analysis, stage, surgery and adjuvant chemotherapy remained statistically significant predictors. Medicaid vs. Private Insurance also remained a significant detriment of survival (OS: HR=3.6, p&lt;0.0001; DFS: HR=2.8, p&lt;0.0001) as did having No Insurance for OS (HR=2.0, p=0.0074). Conclusions: To our knowledge, this is a largest retrospective study showing type of insurance to be a strong predictor of outcome in this very specific breast cancer subtype which remained significant after adjusting all other variables. Further insight is needed to determine the cause of this finding.

Black race as a prognostic factor in triple-negative breast cancer patients treated with breast-conserving therapy: a large, single-institution retrospective analysis

Triple-negative breast cancer (TNBC) disproportionately affects black women. However, black race as a prognostic factor in TNBC has not been well studied. We evaluated the effect of race, among other variables, on outcomes in women with TNBC. A total of 704 patients with stages I-III TNBC treated with breast-conserving surgery ± adjuvant radiation therapy (RT) and chemotherapy were identified from an institutional database. Competing risk analyses, Kaplan-Meier methods, and Cox proportional hazards models identified associations among clinicopathologic variables on locoregional recurrence (LRR), distant recurrence (DR), and overall survival (OS). LRR was defined as a biopsy proven, triple receptor-negative recurrence in the ipsilateral breast or regional lymph nodes. At a median follow-up of 51 months, there were 55 LRR, 61 DR, and 111 death events. Compared to non-black women, black women had higher disease stage and were more likely to receive axillary lymph node dissection, chemotherapy, and nodal irradiation (all P < 0.05). After adjustment for stage, age, lymphovascular invasion, chemotherapy, and RT on multivariate analysis, black race was prognostic for increased risk of LRR (hazard ratio [HR] = 3.17; 95 % confidence interval: 1.7-5.8; P = 0.0002). The 5-year risk of regional recurrence was higher in black women (10 vs. 2 %, P < 0.0001), but local failures were similar between groups (3.0 vs. 5.3 %, P = 0.15). RT was an independent predictor for decreased LRR and increased OS on multivariate analyses (P = 0.0006 and P = 0.0003, respectively). Black women with TNBC had equivalent local control, but higher risk of regional nodal failure, compared with non-black counterparts. The routine use of comprehensive nodal irradiation may be beneficial for black women with TNBC.

Vimentin as a poor prognostic factor for triple-negative breast cancer

Triple-negative breast cancer (TNBC), characterized by the absence of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2, is a highly heterogeneous disease. Recent studies suggest that there are links between TNBC and the epithelial-mesenchymal transition (EMT). To identify prognostic biomarkers and novel therapeutic targets, vimentin, one of the most major factors associated with EMT was investigated in TNBC. Sporadic invasive ductal carcinoma specimens were obtained from 659 Japanese patients, and 90 (14 %) cases were diagnosed as TNBC. The vimentin mRNA and protein expression levels were evaluated by quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry. The mRNA expression of vimentin was significantly upregulated in the basal-type breast cancer cell line. Immunohistochemically, the vimentin expression was significantly higher (p = 0.0042) in TNBC compared with the other subtypes. Vimentin expression was associated with a younger age (p = 0.016), high nuclear grade (p = 0.023) and high Ki67 expression (p < 0.0001), and a poorer prognosis in terms of both the recurrence-free survival (RFS) (p = 0.0058) and overall survival (OS) (p = 0.013) in TNBC patients. A multivariate analysis showed that vimentin expression was an independent prognostic factor for the RFS (p = 0.043). Vimentin expression was also associated with a significantly shorter RFS (p = 0.021) and OS (p = 0.017) in patients with basal-like breast cancer (BLBC). The elevated expression of vimentin contributes to the aggressive phenotype and poor prognosis in TNBC. Vimentin expression might be useful as a biomarker for the prognosis of TNBC.

RAD51 genotype and triple-negative breast cancer (TNBC) risk in Polish women

The most lethal damage for the cell among all damage is double-strand breaks (DSB) of DNA. DSB cause development of cancer diseases including the triple-negative molecular subtype of breast cancer. The aim of this work was to evaluate the single nucleotide polymorphism -135G>C (rs1801320) of the RAD51 gene encoding DNA repair proteins by homologous recombination (HR) in triple-negative breast cancer (TNBC). We assessed the RAD51 -135G>C polymorphism in 50 women with triple-negative breast cancer and in 50 women from the control group. RAD51 polymorphism was analysed by the PCR-RFLP (restriction fragment length polymorphism) technique. Our results demonstrated a significant positive association between the RAD51 C/C genotype and TNBC, with an adjusted odds ratio (OR) of 5.95 (p = 0.002). The homozygous C/C genotype was found in 68% of breast cancer cases and 20% of controls. The variant 135C allele of RAD51 increased TNBC risk. This is the first study linking single nucleotide polymorphisms of the RAD51 gene with TNBC incidence in the population of Polish women. In conclusion, RAD51 polymorphisms may be regarded as predictive factors of triple-negative breast cancer in the female population. Large studies are needed to confirm our findings.

Abstract P1-06-03: Microvessel density as determined by computerized image analysis of CD34 and CD105 expression correlates with poor outcome in triple-negative breast cancer

Abstract Background: Triple-negative and basal-like breast cancers tend to relapse and metastasize early with a peculiar pattern of dissemination. Occurrence of metastasis is related to angiogenesis, yet literature on angiogenic characteristics of these tumors is limited. We examined blood vessels topography and microvessel density (MVD) in triple-negative breast cancer (TNC), investigating their association with clinicopathological criteria, basal phenotype and prognosis. Methods: For identification of blood vessels, FFPE whole tissue sections were stained with CD34 and CD105. Slides were scanned using ScanScope, Aperio Digital Pathology System. Subsequently, digital slides were analyzed with Aperio ImageScope, Microvascular Density v1 algorithm. Total MVD and MVD in three areas (intratumoral, IT, peripheral tumoral, PP and peritumoral, PT) were assessed as described by Weidner et al and correlated with prognosis. Results: A total of 48/133 pts (36.1%) were younger than 50 years (mean 55 yrs; range 32–86 yrs) and 41 (30.8%) were premenopausal women. The majority of TNCs were invasive ductal carcinomas NST (106/133); 80 (60.2%) cases were lymph node positive and 101 (75.9%) were found to have a basal-like phenotype. LVI was identified in 64/133 (48.1%) tumors. Median follow-up was 40 mos (range 1–161 mos). Overall, 53 (39.1%) patients had developed a recurrence by the time of the last follow-up, 49 (36.8%) had developed distant metastasis and 45 (33.8%) had died from breast cancer. Blood vessels were located largely in the peripheral tumoral area. Tumors with high total CD34 MVD, CD34 MVD at the peripheral area (PP-MVD) and CD34 MVD at the peritumoral area (PT-MVD) were significantly associated with the development of visceral metastasis (p = 0.006; p = 0.013; p = 0.019). Also, high CD34 PT-MVD was associated with recurrence (p = 0.010), advanced tumoral stage (p = 0.028) and lymph node (LN) metastasis (p = 0.048). While high total CD105 MVD and CD105 PP-MVD were associated with distant metastasis (p = 0.048; p = 0.031), CD105 PT-MVD was correlated with vascular invasion (p = 0.015). Moreover, CD105 PP-MVD was strongly associated with poorer overall survival (OS; p = 0.001) and disease-free interval (DFI; p = 0.001). Interestingly, TNCs with a basal phenotype showed higher CD34 PT-MVD and CD105 PT-MVD (p = 0.017; p = 0.046). In multivariate analysis for OS, total CD34 MVD (RR = 2.20; 95% CI 1.11–4.35; p = 0.023) and CD105 PP-MVD (OS: RR = 2.78; 95% CI 1.41–5.49; p = 0.003) retained significance after adjustment for stage, LN status and distant metastasis. As for DFI, CD34 PT-MVD (RR = 1.99; 95% CI 1.03–3.87; p = 0.040) and CD105 PP-MVD (RR = 2.90; 95% CI 1.54–5.48; p = 0.001) showed statistical significance after adjustment for the same variables. Conclusions: Our findings demonstrate significant associations between MVD in different tumoral areas and various parameters in triple-negative breast tumors, indicating that TNCs with aggressive features have higher MVDs. Furthermore, peripheral tumoral CD105 MVD is a strong, independent prognostic factor in TNC. Finally, blood vessels are mainly located in the peripheral area of TNCs and seem to have a critical role in tumor progression by representing means for disease dissemination. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-06-03.