Neu Status Research Articles

Frequency of recurrence in early breast cancer and association of known prognostic factors with relapse and time to relapse in a tertiary care hospital in Pakistan.

e21175 Background: Breast cancer is the most prevalent cancer in women and a leading cause of cancer-related death. Following surgery and initial treatment, recurrence remains a risk for all patients with breast cancer. Methods: This is a cross sectional study. Complete data of 300 patients treated for early breast cancer during 2004-2008 at Oncology Dept, LNH was analysed. Out of these 300 patients, 76 patients had relapsed. Data were collected on SPSS version 17, and analyzed. Variables were age, tumor size, nodal status, tumor grade, hormonal status and her 2 neu status, taxanes versus non taxanes treatment, use of trastuzumab and hormonal treatment. Early relapse was defined as in less than 2 years and relapse after 2 or more years was taken as late relapse. Results: 300 patients were analyzed. Mean age of our patients were 47.6 ± 11.2 years. Out of these 300, 73 patients relapsed (24.7%). 44 patients (58%) relapsed in less than 2 years, and 33 patients (42%) relapsed after 2 years. Risk factors associated with relapse included tumor size, nodal status, grade and hormone receptor status. There was significant association between hormonal status and time to relapse ((P 0.001). We did not find any significant association in over all relapse rate, and different immunohistochemical subtypes but on analyses of time to relapse, early relapse was higher in triple negatives and her 2 neu positive group (P 0.001). There was a trend towards increased incidence of visceral and brain metastases in early relapse and of bone metastases in late relapse. Patients who relapsed early (< 2 years) had median survival of 1 year while those who relapsed after 2 years had median survival of 4 years. Conclusions: We found strong association of ER/PR status only with time to relapse in carcinoma breast. Early relapse was higher in triple negatives and her 2 neu positive group with shorter median survival in early relapse.

Racial variation of leptin levels in women with breast cancer.

1590 Background: Incidence of breast cancer is lower in African-American women than White women, but African-American women have worse survival and represent a greater proportion of young women. Obesity is a factor in disparities. Leptin and adiponectin, hormones produced by adipose tissue, are active in breast cancer cell lines and correlate with tumor characteristics. Leptin has mitogenic effects in vitro and increases with obesity; adiponectin has inhibitory effects and decreases with obesity. Studies suggest that adipocytokine levels differ by race, adjusted for body mass index (BMI). In a small study of adipocytokine levels by race in women with breast cancer, we found that African-American women had higher levels of adipocytokines, adjusted for BMI. There was also a correlation between tumor grade and adipocytokine levels. In this study, we examined a larger number of women to replicate these findings. We measured adipocytokine levels in African-American and White women with breast cancer and correlated tumor characteristics with adipocytokine levels. Methods: We performed a retrospective case-case study of 164 African-American and 172 White breast cancer patients. Adiponectin and leptin levels were measured in serum by ELISA. Records were abstracted for BMI, age, stage, grade, ER/PR, and Her-2 neu status. Adipocytokine levels were compared between races by Chi-square analysis and adjusted for BMI. Results: Mean leptin levels were higher in African-American women, adjusted for BMI (p<0.001). There was a trend toward triple negative tumors with higher leptin levels (p==0.06). There was a correlation between younger age and premenopausal status with lower adiponectin levels, adjusted for BMI. Women aged 60 had higher adiponectin levels than women aged 40-49, (p==0.002). Premenopausal women had lower adiponectin levels (p==0.01). Conclusions: Our findings suggest that leptin levels may differ by race in women with breast cancer, adjusted for BMI. Higher levels of leptin may correlate with triple negative tumors. We found that younger women have lower levels of adiponectin, which may play a role in breast cancer in young women. With further study, differences in leptin may prove to be a biologic cause of some of the observed disparities in breast cancer.

Expression of Epidermal Growth Factor Receptor Tyrosine Kinase Family in Fine Needle Aspiration and Permanent Specimens of Invasive Lobular and Ductal Breast Cancers

Recently, the role of HER-2/Neu gene amplification has been enthusiastically investigated in breast cancer. Determining the HER-2/Neu status could be achieved by evaluating either histologic samples or cytologic specimens obtained by Fine Needle Aspiration (FNA). This study aimed at determining the concordance of HER-2/Neu expression in FNA and histologic sections. FNA samples, as well as their corresponding histologic sections of 90 cases with breast cancer were evaluated in Tabriz Sina Teaching Center in a 13-month period of time. The immunohistochemistry was employed for determining the HER-2/Neu amplification for both methods. The concordance rate and agreement were determined between the two methods. Ninety specimens of women with a mean age of 50.93 +/- 10.64 (29-84) years were assessed. There were 84 cases with invasive ductal carcinoma and 6 cases with invasive lobular carcinoma. Lymph nodes were involved in 50 cases and there were vascular and neural involvement in 40 and 35 cases, respectively. Her-2/Neu was not detected in 27 cases (30%) with weak and strong amplifications in 47 (52.2%) and 16 (17.8%) cases of FNA specimens, respectively. Her-2/Neu was not detected in 29 cases (32.2%) with weak and strong amplifications in 42 (46.7%) and 19 (21.1%) cases of histologic specimens, respectively. The concordance rate was 70% between the two methods. The agree ment was statistically significant between the two methods, as well (kappa = 0.51, p < 0.001). HER-2/neu gene amplification can be reliably estimated by immunohistochemistry on breast cancer FNAs and a good correlation has been found between this and results on histological sections.

Cancer biomarker HER-2/neu in breast cancer in Indian women

In Indian women with breast cancer, the HER-2/neu gene is amplified in 30% of cases. Elevated serum HER-2/neu levels have been shown to be associated with a poor clinical prognosis and decreased survival in early stage breast cancer patients, and testing for this may help to manage the disease. The present study was therefore to estimate serum HER-2/neu levels in breast cancer patients and associate these with other prognostic factors. Serum HER-2/neu levels were studied in 207 patients with breast cancer, 15 with benign breast diseases, (BBD) and 175 age-matched healthy controls. Patients' age, menopausal status, node, and estrogen receptor (ER) and progesterone receptor (PgR) status were compared with serum HER-2/neu levels. Serum HER-2/neu overexpression was associated with age, disease stage and positive nodal status but not with menopausal status. Serum HER-2/neu levels were negatively correlated with hormone receptor positivity. HER-2/neu serum tests should be done more frequently in Indian women with breast cancer, irrespective of their ER and PgR hormone receptor status. ELISA is a reliable and economical tool to assess the HER-2/neu status in tumors, when breast tissue samples are not available.

Analysis of CD40 expression in breast cancer and its relation to clinicopathological characteristics.

Breast cancer is currently the most common cancer in women worldwide. For this reason, new biomarkers for better predicting response to treatment are needed. CD40, described as expressed in haematological and epithelial tumors, is linked to apoptosis and offers promise as a new predictive/ prognostic marker. We evaluated CD40 expression in formalin-fixed, paraffin-embedded samples from 181 breast carcinomas using immunohistochemical staining with CD40 antibody. Samples were divided according to hormone (oestrogen receptor /ER/, progesterone receptor /PR/) and her-2/neu status into groups: 1.Luminal A (ER+PR+her-2/neu-), 2. Luminal B (ER+PR+her-2/neu+), 3.Triple-negative (ER-PR-her-2/neu-) and 4. Her-2/neu (ER-PR-her-2/neu+). The results of CD40 staining were correlated with clinicopathological data. CD40 was found to be expressed in membrane, cytoplasm and nucleus. Normal ducts expressed cytoplasmic CD40 in 30% of cases, in breast tumor ducts in 53% of cases. CD40 was evaluated as an independent marker and significant positive correlation was found with Bcl-2 (p =0.002), early stage (p =0.016) and preoperative chemotherapy (p =0.043). There was higher overall survival for patients with cytoplasmic CD40 expression (0.05). Differences in expression of cytoplasmic CD40 between groups with different hormonal and her-2/neu status were statistically highly significant (p=0.00003). In groups with different hormonal status, a positive statistical correlation was found for the luminal A group with relapse (p=0.024) and stage (p=0.006). No correlation was found with age, disease onset, family history of cancer/ breast cancer, patient history, hormonal replacement therapy, menopausal status at onset of disease, adjuvant chemotherapeutic treatment or disease free survival. Nuclear expression of CD40 was found to be unrelated to any clinicopathological data. However, there was higher ratio of positive cases in cancer cases (83%) than in normal tissue (30%). In conclusion, cytoplasmic expression of CD40 is related to factors connected to better prognosis and suggest that CD40 may have potential as a new prognostic factor in breast cancer.

Immunohistochemical detection of Her-2/neu overexpression in breast carcinoma in Nigerians: A 5-year retrospective study

To determine the pattern of Her-2/neu status among breast carcinoma in the University of Benin Teaching Hospital, Benin City, Nigeria. Immunohistochemical staining for Her-2/neu was performed on 10% formalin-fixed, paraffin-embedded primary carcinoma of the breast from 83 patients, between 2003 and 2007 using anti-Her-2/neu rabbit polyclonal antibody (DakoCytomation, CA, USA) and reactivity detected by an avidin-biotin immunoperoxidase method. The clinicopathologic parameters analyzed were patients' age, histological types, and tumor grade. The Her-2/neu Dako scoring system was used. During the 5-year period, 83 histologically confirmed cases of invasive breast carcinoma were assessed for Her-2/neu status. These included 67 (80.7%) cases of invasive ductal carcinoma, nine cases of invasive lobular carcinoma (10.8%), three cases of medullary carcinoma (3.6%), two cases of papillary carcinoma (2.4%), and a case each of mucinous and clear cell carcinoma (1.2%). Mean age of patients was 48.9 ± 13.6 years with an age range of 25 to 83 years. Only nine (10.8%) cases showed immunopositivity for Her-2/neu. Seven of these were cases of invasive ductal carcinoma, while two were invasive lobular carcinoma. Tumors were predominantly high grade with grade III, II, and I accounting for 42, 28, and 13 cases, respectively. There was no significant correlation between patient age, the various histological types, tumor grade, and Her-2/neu positivity. This study indicates that Her-2/neu may not be useful in defining the prognosis of breast cancer in this environment. Nevertheless, it highlights the need for routine immunostaining for Her-2/neu before institution of trastuzumab (Herceptin) therapy.

Status of HER-2/neu receptors and Ki-67 in breast cancer of Indian women

Background:Breast cancer is a leading cause of death in women. Receptor status is the most important prognostic and predictive marker for breast cancer.Aims:The present study was conducted with an aim to analyze breast cancer of Indian women with discordant receptor status, probably hormone dependent estrogen receptor (ER) positive, progesterone receptor (PgR) negative or ER– negative and PgR+ positive subgroup profile, infiltrating ductal breast cancer (IDC) not otherwise specified.Materials and Methods:Specimens from 100 IDC were grouped into three categories according to hormonal status (group 1: ER+ positive and PgR+ positive, group 2: ER+ positive and PgR– negative or ER– negative and PgR+ positive, group 3: ER– negative and PgR– negative) evaluated prognostic parameters.Statistical Analysis:Statistically significant difference was found between tumor receptor status distribution and menopausal status (P = 0.0235), age of patients (P < 0.001), histopathologic grade (P < 0.001), vascular invasion (P = 0.006), HER-2/neu status (P = 0.004) and Ki-67 proliferation rate (P < 0.001).Results:Group 1 tumors were found exclusively in post-menopausal patients with average age 68.9 years, most of which had intermediate grade II, without vascular invasion, with HER-2/neu status score predominantly 0 or 1+ and lower Ki-67 proliferation rate. Group 2 tumors were found predominantly in younger post-menopausal patients with average age 57.5 years, with vascular invasion found in 23% of cases. Group 3 tumors mostly had higher histopathologic grade, showed the highest percentage of the Ki-67 positive tumor cells and vascular invasion in 30% of the cases.Conclusion:It is concluded that patients with group 2 breast cancer were younger post-menopausal women, with tumors moderately differentiated, HER-2/neu score 0 or 1+ and with lower Ki-67 proliferation rate.

Correlation of her 2/neu gene amplification with immunohistochemistry and other prognostic factors in breast carcinoma

The purpose of this study was to determine relationship between HER-2/neu status and estrogen receptor, progesterone receptor, grade and age by comparing fluorescence in situ hybridization and immunohistochemistry. One hundred invasive breast carcinomas were reviewed and fluorescence in situ hybridization analysis was performed in all cases. Immunohistochemical scores showed a perfect concordance with fluorescence in situ hybridization amplification ratios (p < 0.0001). The results indicated a significant correlation between HER-2/neu and grade, but an inverse relationship between HER-2/neu and hormone receptors. In women aged ≤ 45 years, an inverse relationship between HER-2/neu and progesterone receptor was found and no association was noted between HER-2/neu and other factors. In women aged > 45 years, the results indicated a significant correlation between HER-2/neu and grade, and there was an inverse relationship between HER-2/neu and hormone receptors.

Relationship of Polymorphisms of Glutathione S-Transferase GSTT1 and GSTM1 With the Response to Chemotherapy In Mexican Women with Advanced Breast Cancer

Introduction: Breast cancer is a common disease diagnosed in Mexican women and the first leading cause of death [1]. Heterogeneity in patients’ response to treatment is consistently observed across populations. Glutathione S-transferases (GSTs) are involved in the metabolism of environmental carcinogens, reactive oxygen species and chemotherapeutic agents by catalyzing the glutathione with electrophilic compounds. The deletion of GSTT1 and GSTM1 genes result in loss of enzyme activity. A few studies evaluated the response to treatment and the polymorphisms of GSTT1 and GSTM1. The aim of this work is to make the association of the null polymorphisms of GSTT1 and GSTM1 with the response to chemotherapy basically doxorubicin and cyclophosphamide. Methods: The genotyping of thirty patients with breast cancer was made with the Polymerase chain reaction, to identify the polymorphisms of GSTT1 and GSTM1. We determine the status of Her-2 neu, estrogen and progesterone receptors, then the response to treatment was made with an ultrasound and pathological data. We made the association with the χi2 statistics using a p≤0.05. Results: Using the Sigma Stat 3.5 program and the chi-squared analysis, we do not observe a significant association with the GSTT1+/GSTM1+, GSTT1-/GSTM1+ and GSTT1-/GSTM1-polymorphisms and the better or worse response to cyclophosphamide and doxorubicin. With the Her-2 neu, estrogen and progesterone receptors status, we neither found an association with the response to the therapy. Conclusion: This study suggests that GSTT1 and GSTM1 polymorphisms have no statistical significance between the genotype of women with advanced breast cancer and the response to neoadjuvant chemotherapy, but we can see a clear tendency toward better response with the null genotype.

Her-2 neu (Cerb-B2) expression in fine needle aspiration samples of breast carcinoma: A pilot study comparing FISH, CISH and immunocytochemistry

Background:Breast cancers with Her-2 neu gene amplification are recognized as important markers for aggressive disease and targets which respond to therapy with trastuzumab. Her-2 neu testing on histological sections is routinely performed to select patients who may benefit from anti- Her-2 neu therapy. Few reports are available which document Her-2 neu status on fine needle aspirates (FNA).Aim:This pilot study is to document expression of Her-2 neu (Cerb-B2) on cytospin smears from FNA of patients with breast carcinoma.Materials and Methods:Twenty samples of FNA already collected for diagnostic purposes from patients with primary breast carcinoma were studied for demonstration of Her-2 neu expression by immunohistochemistry (IHC), Fluorescent in-situ hybridization (FISH) and chromogenic in-situ hybridization (CISH) on cytospin smears from FNA. Their expression was compared with tissue sections where possible.Results:Good correlation was observed between Her-2 neu protein expression and gene amplification in cytospin smears. Three of five (60%) breast carcinomas cases with 2+ and 3+ staining on IHC showed gene amplification by FISH and CISH. Three of 7 (43%) and 5 of 7 (71%) cases negative/1+ staining on IHC did not show gene amplification by FISH and CISH respectively. Tissue sections from 10 cases with 2+ and 3+ staining for Her2neu by IHC showed gene amplification in 8 cases.Conclusion:Demonstration of Her-2 neu by IHC, FISH or CISH in FNA is possible and may play a role in the management of patients with advanced breast cancer or those cases where surgical resection is not advisable.

Assessment of HER2/Neu status by fluorescence in situ hybridization in immunohistochemistry-equivocal cases of invasive ductal carcinoma and aberrant signal patterns: A study at a tertiary cancer center

HER-2/neu status determines the eligibility for targeted therapy with trastuzumab in breast carcinoma. Evaluation for HER-2/neu protein expression by immunohistochemistry (IHC) and gene amplification by fluorescence in situ hybridization (FISH) has become the gold standard. Since data on HER-2/neu assessment by IHC and FISH and studies regarding concordance between the results of the two techniques are limited, especially from India, we sought to study HER-2 gene amplification status by FISH in equivocal (2+) cases by IHC and also study aberrant signal patterns. Mastectomies and breast core biopsies, equivocal for HER-2/neu protein expression, were analyzed for HER-2 amplification by FISH. IHC (DAKO) and FISH (PathVysion dual-probe system) tests were performed on 68 of 112 (after exclusion) 10% neutral buffered formalin (NBF)-fixed paraffin-embedded tissues and evaluated according to American Society of Clinical Oncology ASCO guidelines. Chi-square (χ2) test and the two-tailed P value were applied using Graphpad Quickcels software, version 2006. It was found that 73.5% of the IHC 2+ patients were negative for HER-2/neu amplification, 25% were positive (ratios ranging from 2.3 to 5.6) and 1 patient was equivocal (2.2). Retesting FISH HER-2 equivocal case on another tumor block by IHC demonstrated HER-2 overexpression of protein 3+, thus resolving the equivocal status. Polysomy and HER-2 genetic heterogeneity were seen frequently. The findings reiterate that IHC HER-2 equivocal cases are a heterogeneous group and need FISH for further categorization. Low concurrence (25%) rate between both IHC and FISH results in the equivocal scenario can be attributed to tumors with polysomy 17 and HER-2/neu genetic heterogeneity.

Abstract P3-10-17: The Fractal Analysis of Tumor Histology Is an Independent Prognostic Factor in Breast Cancer Patients

Abstract Background: Despite the usefulness of standard clinical prognosticators for breast cancer patients, recommendations for systemic adjuvant therapy are not entirely straightforward. Hence, there is a need for a more accurate independent prognosticator that provides additional information to improve risk assessment and associated therapeutic decisions. Methods: We used fractal dimension analysis to quantitatively assess the morphologic complexity of breast epithelium. To investigate the relationship between fractal dimension and patient survival we applied fractal analysis to pan-cytokeratin stained tissue microarray (TMA) cores derived from a series of 408 patients with over 10 years follow up, and analyzed three TMA cores for each patient. A data-oriented approach was used to stratify patients according to low (&amp;lt;1.56), intermediate (1.56-1.75), and high (&amp;gt;1.75) fractal dimension. Univariate statistical analyses were performed to show the relationship between outcome and fractal dimension, tumour size, tumour grade, lymph node status, estrogen receptor status, and HER-2/neu status. Multivariate analysis was performed to assess the relative effect of these prognosticators on disease-specific and overall survival. Results: Patients with higher fractal score had significantly lower disease-specific 10-year survival (69.4%, 56.4%, and 25.0%, for low, intermediate, and high fractal dimension, respectively, P&amp;lt;0.001). Overall 10-year survival showed a similar association with fractal dimension. Cox regression analysis showed fractal dimension, lymph node status, and grade to be the only significant (P&amp;lt;0.05) independent predictors for both disease-specific and overall survival. Fractal dimension had the highest hazard ratio for overall survival 2.7(95% confidence interval (CI)=1.6-4.7); P&amp;lt;0.001), and the second highest for disease-specific survival 2.6(95% CI=1.4-4.8; P=0.002) versus 3.1(95% CI=1.9-5.1; P&amp;lt;0.001) for lymph node status. Discussion: Except for lymph node status, morphologic complexity of breast epithelium as measured by fractal dimension is more strongly and significantly associated with disease-specific and overall survival than standard clinical prognosticators. Furthermore, it is independent of standard prognosticators, and unlike tumour grade, it provides prognostic information that can be objectively assessed from TMA cores. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-17.

Abstract P4-09-12: The Effect of Age on Breast Cancer Outcomes in Women with Her-2 Positive Breast Cancer: Results from the HERA Trial

Abstract Background: Previous research has suggested that young age at diagnosis is an independent risk factor for breast cancer recurrence and death. However, young women are more likely to develop more aggressive subtypes of breast cancer and no prior studies have adequately controlled for Her-2/neu status or anti-Her-2 treatment. We sought to evaluate whether age is a prognostic factor in the HERA trial, which is a large randomized phase III trial of women with Her-2 positive early stage breast cancer who were randomized to receive trastuzumab (for one or two years) or observation following (neo)-adjuvant chemotherapy. Methods: We used 2 year median follow-up data from the HERA trial and conducted Cox proportional hazards models. Age was dichotomized at ≥40 years versus &amp;gt; 41 years to evaluate its prognostic effect on outcomes in women who received trastuzumab and those who did not. Only patients included in the 1-year trastuzumab and observation arms were included in the present analysis. Results: Of the 1703 women randomized to one-year of trastuzumab and 1698 to observation, 722 (21%) were age 40 or younger at study entry. Younger women were more likely to have node negative (33% vs 37%), estrogen receptor (ER)-negative (56 % vs 49%), and progesterone receptor (PR)-positive disease (41% vs 32%). In separate Cox models controlling for T and N stage, grade, ER and PR status, chemotherapy, endocrine therapy, and menopausal status at randomization, disease-free survival (DFS) hazard ratios were consistent for women ≥40 years compared with &amp;gt; 41 years old patients, regardless of whether they were assigned to observation or to receive trastuzumab. DFS were as follow: observation group: HR (younger/older) = 1.09, p-value = 0.62; trastuzumab group: HR (younger/older) = 1.05, p-value = 0.84. There was no statistically significant interaction between age and treatment (p=0.94). Conclusions: In women with early stage Her-2/neu positive breast cancer in a large RCT, age was not an independent predictor of risk of early recurrence. Future research to investigate whether age is a predictor of later risk of recurrence or is a predictor among women with other tumor subtypes is warranted. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-09-12.

Abstract P4-09-16: WAVE3 Over-Expression Is Associated with Adverse Tumor Characteristics and Mortality in Breast Cancer

Abstract Background: WAVE3 regulates actin polymerization and subsequent cell migration leading to enhanced metastatic potential. Based on pilot data that suggested WAVE3 expression was associated with high histologic grade and absence of estrogen receptor (ER) expression we hypothesized that WAVE3 expression would correlate with ER status and tumor grade in a matched group of breast cancer (BC) patients. WAVE3 expression was alsoanalyzed in relation to adverse tumor characteristics, distant recurrence (DR) and BC specific mortality. Methods: Our institutional BC database was reviewed for patients who presented with, invasive BC from 1999-2009. Matching by stage and treatment was achieved for 61 patients with Scarff-Bloom-Richardson (SBR) grade 1 and ER+ tumors (SBR1/ER+) to 61 patients with SBR grade 3 and ER-tumors (SBR3/ER-). Cytosolic WAVE3 tumor expression was determined by immunohistochemistry. The product of stain intensity (0-3) and percentage of cells staining (0-100) was used to derive a WAVE3 score (0-300). The log rank test was utilized to compare BC specific mortality or distant recurrence free survival at various WAVE3 scores. A score of ≥212 was found to have the strongest association with poor outcome. The association between WAVE3 score and clinicopatholigic features, DR and BC specific mortality was assessed. Results: Increased frequency of Her2-neu (+) status, DR and BC specific mortality was noted in the SBR3/ER-group but WAVE3 score was no different between the two groups(Table1). In all 122 patients median WAVE3 score increased with tumor size (0.234, p=0.009), (+) lymph node status 200 vs. (-), 145, p =0.03, and stage (I, 160 vs. II,180 vs III, 240, p=0.012). There was no association between WAVE3 score and Her2-neu status(+200 vs. -180, p=0.51). In the SBR1/ER+ group only (+) lymph node status remained associated with WAVE3 score(+) 200 vs. (-) 130 (p=0.02). In the SBR3/ER-group only lymph node status lost association with WAVE3 score (+)180 vs.(-)170, (p=0.50). DR and BC specific survival could only be assessed in the SBR3/ER-group. Median WAVE3 score was elevated with DR (240 vs. none, 160, p=0.03) and BC specific mortality (270 vs. none 170, p=0.004). A WAVE3 score ≥212 was associated with distant recurrence and BC specific mortality on Kaplan Meier analysis (p=0.01) and P&amp;lt;0.001). On multivariate analysis a WAVE3 score ≥212 was associated with an increased risk for BC specific mortality (p=0.009). The association of DR and WAVE3 score ≥212 approached significance (p=0.068). Table 1 Conclusion: WAVE3 expression is not associated with tumor grade, and ER or Her2 neu status. WAVE3 is associated with tumor size, stage, DR and BC specific mortality in the high risk SBR3/ER-group. A WAVE3 score of ≥212 is associated with distant recurrence and breast cancer specific mortality on univariate analysis and BC specific mortality on uni-and multivariate analysis. WAVE3 expression may contribute to adverse outcome in high risk breast cancer patients. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-09-16.

Androgen receptor expression in breast cancer patients tested for BRCA1 and BRCA2 mutations

To assess the expression of receptors for androgen (AR), oestrogen (ER) and progesterone (PR) as well as human epidermal growth factor receptor type 2 (Her-2/neu) status of breast carcinomas in breast cancer susceptibility gene (BRCA) BRCA1/2 mutation carriers and BRCA1/2 negative tested women. One hundred and thirty-five breast cancers in women tested for BRCA1/2 mutations. Screening for BRCA1 and BRCA2 mutations was performed by direct sequencing of all BRCA1 and BRCA2 exons as well as the surrounding intronic sequences. Additionally, BRCA genes were analysed with multiplex ligation-dependent probe amplification. Consecutive paraffin sections were examined immunohistochemically for AR, ER, PR and Her-2/neu. Of the 135 tumours, 43 (32%) were BRCA1-related, 18 (13%) were BRCA2-related and 74 (55%) were BRCA1/2-negative. Seventy-two per cent of the BRCA1-related, 22% of the BRCA2-related and 12% of the BRCA1/2-negative tumours were triple (ER, PR, Her2neu)-negative. Eighty-four per cent of BRCA1 mutated cancers were high-grade (G3) tumours. ARs were expressed in 30% (13 of 43) of BRCA1-related, in 78% (14 of 18) in BRCA2-related tumours and in 76% (56 of 74) in BRCA1/2 negative tumours. Twenty-one per cent of ER-negative BRCA1-related tumours expressed androgen receptors. Approximately one in five BRCA1 mutated breast cancers negative for ER and PR express androgen receptors. Modulation of AR might open a new avenue for treating these high-risk cancers.

Clinicopathological Characteristics and Her-2/neu Status in Chinese Patients with Uterine Papillary Serous Carcinoma

Objective. To analyze clinico-pathological features of Chinese patients with UPSC, and investigate roles of Her-2/neu protein expression and gene amplification in UPSC prognosis. Methods. Thirty-six patients with UPSC treated in Cancer Hospital of Fudan University from 1996 to 2006 were analysed retrospectively. Chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) were performed to evaluate Her-2/neu gene amplification and protein expression respectively. Results. The median age was 63 years, and 61% (22/36) were late stages (stage III/IV). The 1-year, 3-year, and 5-year overall survival (OS) was 73.1%, 51.9% and 43.9%, respectively. Advanced stages (P = .0006) and deep myometrial invasion (P = .0138) were significantly associtated with a shorter OS. In 36 cases, 27.8% (10/36) showed 2+ staining and 8.3% (3/36) showed 3+ by IHC. Amplification of the Her-2/neu gene was observed in 11.1% (4/36) cases. The 5-year overall survival rate in Her-2/neu IHC 2 + ∼3+ and 0 ~ 1+ cases was 12.9% and 68.6% respectively. Her-2/neu protein expression 2 + ∼3+ was significantly associated with advanced surgical stage and worse overall survival (P = .03 and P = .0023, resp.). Conclusion. Chinese patients with UPSC showed characteristics of deep myometrial invasion, advanced stages and poor overall survival. Her-2/neu protein overexpression is associated with advanced stage and poor survival outcome.

HER-2/neu overexpression in breast cancer

Background: In breast carcinoma, amplification &amp;/or over-expression of HER-2/neu has been associated with a group of unfavorable prognostic factors. The Food &amp; Drug Administration Agency approved Trastuzumab (Herceptin) for the therapy of metastatic breast cancer but only in patients with amplification &amp;/or over-expression of this gene. Because of these advances, evaluation of HER-2/neu status in breast cancer specimens is of vital importance.Patients and methods: thirty eight females with breast carcinoma were included in this study. All histologic sections were stained routinely with the hematoxylin and eosin stains andimmunohistochemically for HER2/neu. All cancers were graded and subcategorized into ductal and lobular carcinomas, with or without an insitu component, and with or without Paget disease. The assessment of HER-2/neu over-expression was quantitated.Results: Of the twenty four cases of infiltrative duct carcinoma (IDC), HER-2/neu staining was positive in ten (41.7%) cases. None of the fourteen cases of infiltrative lobular carcinoma (ILC)showed a positive reaction. Statistical analysis revealed no significant differences in respect to HER2/neu expression between IDC with and without ductal carcinoma in situ (DCI). All cases of IDC with overlying Paget disease (5 cases; 13.2%) were positive for the stain. All grade I cancers were negative for HER-2/neu over-expression, in contradistinction to a positive staining in 41.7% grade II tumors and 62.5% grade III tumors.Conclusion: there is significant correlation between HER2/neu overexpression and cancer histopathological type. The presence or absence of an in situ component had no effect on the marker overexpression. Significant correlation was observed between HER-2/neu overexpression and higher histological grades of IDC. No correlation was found between the positivity of the marker and axillary lymph node status. Similarly, no association was found with the other variables except the size of tumor. It is recommended that HER-2/neu assessment to be included as a routine test in the screening program of all breast cancer Iraqi patients.

Abstract B74: Racial variation of adipocytokine levels in women with breast cancer

Abstract Background: The incidence of breast cancer is lower among African-American women than White women; however, African-American women have worse survival, represent a greater proportion of young women with breast cancer, and tend to have more aggressive tumors at diagnosis. A number of social factors contribute to the disparities; however, biologic differences are being explored. Obesity is a risk factor that has a significant effect in breast cancer disparities. Two cytokines produced by adipose tissue, leptin and adiponectin, have been shown to be active in breast cancer cell lines in vitro and to correlate with breast cancer risk and tumor characteristics in epidemiologic studies. Leptin has mitogenic effects on breast cancer cells in vitro and increases with obesity. Adiponectin has inhibitory effects on cancer growth and decreases with obesity. Data from studies of cardiovascular disease and metabolic syndrome suggest that levels of adipocytokines may differ by race, adjusted for body mass index (BMI). We measured levels of leptin and adiponectin in African-American and White women with breast cancer to test our hypothesis that serum levels differ by race. We hypothesized that African-American women would have higher levels of leptin and lower levels of adiponectin, adjusted for BMI. We also correlated tumor characteristics, including grade, age, stage, ER/PR and Her-2/neu status with levels of these adipocytokines to test the hypothesis that more aggressive tumor features would correlate with higher levels of leptin and lower levels of adiponectin. Methods: We performed a retrospective case-case study of 68 African-American and 59 White breast cancer patients. Adiponectin and leptin levels were measured in serum samples by enzyme-linked immunosorbent assay (ELISA). Medical records were abstracted for BMI, age, stage, tumor grade, ER/PR, and Her-2 neu status. Differences in leptin and adiponectin levels between races were assessed using the Kruskal-Wallis test. Linear regression was used to detect differences in levels after adjusting for BMI. Results: Mean levels of leptin and adiponectin were significantly higher in African-American women, compared to White women. Mean leptin levels were 57.8 ng/ml and 30.1 ng/ml in African-American and White women, respectively (p&amp;lt;0.001). Mean adiponectin levels were 24.68 ng/ml and 19.19 ng/ml in African-American and White women, respectively (p=0.006). A persistently significant difference in both leptin and adiponectin between racial groups remained after adjustment for BMI. Adiponectin levels were positively correlated with higher grade tumors (p=0.04). There was a trend of a positive correlation between leptin and higher grade tumors, but this association did not reach statistical significance (p=0.057). We found no correlation between adiponectin and leptin and stage at diagnosis, ER/PR, Her-2/neu status, orage. Conclusions: Our findings suggest that adipocytokine levels may differ by race in women with breast cancer, adjusted for BMI. There may also be a positive correlation between adipocytokine levels and higher tumor grade in women with breast cancer. Our study population was small, and we will validate the results in a larger sample size. If the results are demonstrated in further studies, differences in adipocytokine levels may prove to be a biologic cause of some of the observed racial disparities in women with breast cancer. Citation Information: Cancer Epidemiol Biomarkers Prev 2010;19(10 Suppl):B74.

Changes of HER2 Status in Circulating Tumor Cells Compared With the Primary Tumor During Treatment for Advanced Breast Cancer

Background HER2/ neu status of tumor cells at metastatic sites in patients with advanced disease may differ from that of the primary tumor. Assessing the presence of target antigens on circulating tumor cells (CTCs) might affect treatment choice. Patients and Methods From June 2007 to October 2008, we collected 23 mL of blood from each of the 76 consecutive patients before and during chemotherapy to determine CTC numbers and HER2 overexpression. CTCs were isolated with the CellSearch System® (Veridex, LLC; Raritan, NJ) and fluorescently stained with the Epithelial Cell Kit®. Tumor Phenotyping Reagent® was used to investigate HER2/ neu overexpression. Results Concordance of HER2 status between the primary tumor and CTCs was 86% (49 out of 57 patients) at baseline and 82% (50 out of 61 patients) in the treatment samples. HER2 overexpression in CTCs was acquired in 8 out of 45 patients (18%) and lost in 3 out of 16 patients (19%) during a treatment containing trastuzumab. The overall discordance rate between the primary tumor and CTCs was 18% (11 out of 61 patients). Patients with HER2 overexpression in CTCs had poorer progression-free survival compared with those without CTCs or with HER2− CTCs (log-rank P =.036). Conclusion Information on the presence or absence of HER2 overexpression can be obtained in CTCs. Larger trials are needed to evaluate the activity of HER2-targeted therapy in patients with acquired HER2 overexpression in CTCs.

Micrometastatic Disease and Isolated Tumor Cells as a Predictor for Additional Breast Cancer Axillary Metastatic Burden

Our study aims were to investigate breast cancer patients with micrometastases or isolated tumor cells (ITCs) in sentinel lymph nodes (SLNs) to determine the rate of non-SLN metastasis and axillary recurrences, and to compare actual non-SLN metastasis rates with those predicted by the Memorial Sloan-Kettering Cancer Center (MSKCC) nomogram. We identified 116 stage I to III breast cancer patients who underwent sentinel lymph node biopsy and had micrometastases or ITCs (<2-mm deposits). Patients underwent completion axillary lymph node dissection (ALND) (group 1) or had no further axillary surgery (group 2). P<0.05 was considered statistically significant. Of 116 patients with micrometastases or ITCs in SLNs, 55 (47%) underwent completion ALND (group 1), and 61 (53%) had no further axillary surgery (group 2). The rate of non-SLN metastases in group 1 patients was 9 (16%) of 55, which was significantly less than that predicted by the MSKCC nomogram (median 30%, P<0.001). Patient age, race, tumor histology, tumor grade, estrogen receptor/Her-2neu status, and lymphovascular invasion did not differ significantly between group 1 patients with positive non-SLNs and those with negative non-SLNs (P>0.05 for each), but patients with positive non-SLNs had larger tumors (P < 0.001). No patient in group 1 experienced an axillary recurrence, while only one patient (1.6%) in group 2 experienced axillary recurrence. The actual rate of positive non-SLNs for breast cancer patients with SLN micrometastases or ITCs who underwent completion ALND was significantly less than that predicted by the MSKCC nomogram. The rate of axillary recurrence is negligible, regardless of the extent of axillary staging.