Neu Expression Research Articles

Correlation of Hormonal Receptors Estrogen Receptor, Progesterone Receptor and Her-2/Neu with Tumor Characteristics in Breast Carcinoma: Study of 100 Consecutive Cases

Introduction-Breast cancer is the most common cancer and leading cause of death in women. In India, its incidence is rapidly rising over last few decades. Present study is aimed to study the pattern of expression of hormonal receptors and Her-2/neu in invasive breast carcinoma and to correlate estrogen receptor (ER), progesterone receptor (PR) and Her-2/neu expressions with various clinicopathological parameters. Material and methods: The present study was carried out in Department of Pathology, S.C.B. Medical College, Cuttack in the year 2013 taking consecutive 100 cases. Routine H&E staining for histological diagnosis and IHC analysis for ER, PR and Her 2/neu was carried out in all 100 cases of breast malignancies. Results: 99% of cases are invasive breast carcinoma, not otherwise specify (IDC-NOS). The age ranges from 23 years to 72 years. Majority of tumors are of grade 2 (70%) followed by grade 3 (30%). ER PR and Her-2/neu expression are seen in 45%, 35% and 30% respectively. Triple negative cases comprise 35%. Higher number of grade 2 tumor shows ER, PR positivity as compared to grade 3 tumors. Her-2/neu expression does not show any significant correlation with age or lymph node status of the patient. Conclusion: ER and PR expression in breast cancers in the current study are found to be comparable to the findings of other authors, but the frequency of HER-2/neu expression is slightly higher. Significant correlation is observed between hormonal receptor status and the grade of the tumor. Inverse relationship is found between Her-2/neu expression and ER, PR receptor status. Her-2/neu expression is increased with size and high grade of tumor.

Correlation between Her2/Neu and Histopatholoical Finding among Sudanese Female Breast Cancer Patients

Background: This study is one of very few studies which have investigated HER-2/neu expression and its correlation with histopathological finding. A total of 328 breast cancer tissue samples had been analyzed using immunohistochemical techniques. Objective: To compare the expression of tumor markers (HER-2/neu) with other prognostic parameters for mammary carcinomas. Results: Stage I- III B breast cancer HER-2/neu negative are (62%), whereas positive are (22%). Stage IV breast cancer HER-/neu negative are (4%), whereas positive are (3%). Recurrent breast cancer HER-2/neu negative are (3%) while positive are (5%).Stage I breast cancer HER-2/neu negative are (10%), whereas positive are (1%). Referring to stage II A, II B breast cancer HER-/neu negative are (59%), whereas positive are (16%). Stage III A, III B breast cancer HER-2/neu negative are (10%), while positive are (4%). HER-2/neu negative breast cancer Tumor size [le] 2 cm are (11%), while positive are (2%). HER-2/neu negative breast cancer Tumor size2-5 cm are (23%), whereas positive are (9%).HER-2/neu negative, Tumor size >5 cm are (26%), while positive are (29%). HER-2/neu negative breast cancer Nodal metastasis 0 are (5%), while positive are (2%). HER-2/neu negative Nodal metastasis1-9 are (16%), whereas positive are (19%). HER2/neu negative breast cancer Nodal metastasis [ge] 10 are (28%), while positive are (30%). HER2/neu negative histological subtype NIDC are (2%), whereas positive are (1%). HER-2/neu negative histological subtype IDC are (67%), while positive are (25%). HER-2/neu negative histological subtype ILC are (1%), whereas positive are (1%). Lastly, HER-2/neu negative with others histological subtype are (3%), whereas positive are (0%). HER-2/neu negative histological grade I are (6%), whereas positive are (2%). HER-2/neu negative histological grade II are (21%), while positive are (7%). Lastly, HER-2/neu negative histological grade II are (40%), whereas

Hormone receptor expression in breast carcinoma at our hospital: An experience

Background: Breast carcinoma is the most common cancer among women in the urban Indian population and second only to cervical cancer in the rural population based on cancer registry data. Prognosis and management of breast cancer are influenced by classic variables such as histologic type, grade, tumor size and lymph node status. More recently hormone receptor (HR) status of estrogen receptor (ER), progesterone receptor (PR), and HER-2/neu expression have opened a new gateway in the field of adjuvant hormonal and/or chemotherapeutic regimen. Objective: The objective of this study was to assess the ER, PR, and HER-2/neu reactivity pattern in breast carcinomas at our hospital. Materials and Methods : A study of 100 patient samples of breast carcinoma was carried out from June 2011 to June 2014 in the Department of Pathology, ESIC Medical College and PGIMSR, ESIC Model Hospital, Rajaji Nagar, Bengaluru. Brief demographic and clinical data were obtained. Immunohistochemistry (IHC) was done by peroxidase antiperoxidase technique for detection of ER, PR, and HER-2/neu receptor status. Details regarding histopathological diagnosis, pathological grading, staging, and HR status of breast carcinoma were collected. Obtained parameters were evaluated using descriptive statistical analysis and presented in terms of percentage. Results : The age of the patients ranged from 24 to 75 years. Majority of tumors were infiltrating ductal carcinomas-not otherwise specified and predominantly histological grade 2. By IHC 52% were ER+/PR+, 25% were HER-2/neu positive and 20% of triple negatives. Conclusion : Detection of hormone expression is of paramount importance since these are one among the classic variables needed for providing suitable adjuvant hormonal and/or chemotherapeutic options, targeted treatment, and predicting prognosis.

Correlation between estrogen receptor, progesterone receptor, HER-2/neu status and other prognostic factors in carcinoma breast in Indian population

Background: Incidence of breast carcinoma (BC) is increasing all over the world. Contrary to the West where it is more common in the elderly; in India it is more common among the younger women. India accounts for nearly six percent of deaths, due to breast cancers in the world and also that one out of every 22 women in India is diagnosed with breast cancer every year. But no information is available on its biological characteristics in the Indian population. Our aim was to evaluate the estrogen receptor, progesterone receptor and HER-2/neu receptor in single institute patients and to compare the expression of these factors with other prognostic parameters such as menopausal state, size of tumor, grade, histological type, mitotic index, lymphovascular invasion and nodal metastases. Methods: This is a prospective study conducted in the Department of General Surgery at Sri Ramachandra Medical University Hospital, Chennai, India. A batch of 60 confirmed patients of BC, diagnosed in between 2012 to 2014, were included in this study. ER, PR and HER-2/neu status were studied by immune-histochemistry and were correlated with other prognostic factors. Results: ER negative and PR negative cases were comparatively high (61.7% and 70%) than the positive cases. 66.7% of the cases were post-menopausal group, 68.3% of cases were T2 tumor and 81.7% cases were ductal carcinoma. There was a significant correlation identified between ER, PR with tumor grade and Mitotic Index of the tumor. HER-2/neu positive cases tended to have significant higher axillary metastasis. There was no significant correlation between ER, PR and HER-2/neu with menopausal state, size, histological type and lymphovascular invasion. Conclusions: The ER, PR and Her2/neu expression in BC in our patients is comparable with the available international data. ER and PR negative expressions are strongly associated with some known bad pathological factors like high grade tumor and high Mitotic Index. HER-2/neu expression was high in advanced diseases like axillary node positive patients.

Study of Her2/Neu Expression in Breast Carcinoma and Correlation with Various Prognostic Parameters

Background: Breast cancer is a heterogeneous disease with varied morphological appearances, molecular features, behavior, and response to therapy. High levels of human epidermal growth receptor 2(HER2/NEU) expressions are associated with recurrence and death in breast cancer (BC). Though it is associated with poorer survival but its main importance is as predictors of response to agents that target this transmembrane protein (transtuzumab). The aim of the study was to evaluate the expression pattern of HER2/NEU in invasive component of breast carcinoma and correlate with various prognostic parameters such a tumour size, histologic grade, histologic type, regional lymph node status, and proliferative marker Ki67. Study Setting & Design: A total of 112 breast cancer cases were studied for the histomorphological features followed by immunohistochemical study in the Department of Pathology of a tertiary care hospital. Material and methods: The lumpectomy or mastectomy specimens were fixed on 10% neutral buffered formalin immediately after surgery in 10 fold volume. Important clinical information from patients were collected in Performa. Finally routine hematoxylin and eosin sections were examined to confirm presence of invasive cancer, ascertain histological types, Histological Bloom Richardson grade (BRG)1 and axillary lymph node status. IHC of HER2/NEU protein was done using HER2/NEU antibody following standard procedures. The tumour was also immunostained with Anti ER, Anti PR, and Ki67 primary antibody. Statistical Analysis: Data was analyzed by SPSS software. Results and observation: The mean age of breast cancer presentation in this study was 44.6 years with standard deviation 11.24.This IHC evaluation for HER2/NEU was positive with 3+ score in 18.75% of cases. The data of HER2/NEU expression profiles were correlated with 3 BRG histologic grades of the tumours, it showed a positive association with high histologic grade. HER2/NEU profiles were inversely correlated with ER, PR expression (p value < 0.05). The HER2/NEU expression profiles was also correlated with tumour size and a significant relationship was found (p = 0.041).HER2/NEU positive tumours were belonged to mostly IDC.HER2/NEU expression profile was correlated with proliferative index Ki67 expression in this study and found a direct relationship (p < 0.005). Lymph node status and HER2/NEU showed a significant statistical correlation (p = 0.037). Discussion: The over-expression of HER2/NEU is associated with poorer prognosis, high grade features and resistant to usual chemotherapy10. Our study also revealed that the Patients with higher levels of HER2/NEU- overexpression or amplification had statistically significant with lower levels of ER and PR positive tumours. We also found that when Her 2 neu was positive in tumours along with ER and PR, then it was mostly high BRG and High Ki67.The new addition from our study wasHER2/NEU expression profile correlation with Proliferative marker Ki67 which revealed a positive association. Conclusion: The expression pattern of HER2/NEU protein showed a strong correlation with high Ki67 expression which means these tumours are likely to have higher proliferative fractions leading to high probability of recurrence without management by herceptin therapy. Keywords: Her2neu, Ki67, Prognostic parameters

Comparative estimation of ErbB family growth receptors, Ki-67 and E-cadherin expression in pancreatic ductal adenocarcinoma cells

Aim. In immunohistochemical research of biopsy and postoperative material of 120 patients with the pancreatic ductal adenocarcinoma the levels of growth receptors of ErbB family and cancer prognostic markers expression were determined.Methods and results. Positive expression of EGFR by the tumour cells is determined in 87,5% of patients with ductal adenocarcinoma, simultaneously 77,5% of such patients had negative, and 13,33% of patients – doubtful HER-status. Expression of Ki-67 inductal carcinoma cells was found in 74,17% of patients, there were distinctions in expression of this marker within one tumour. The decline of expression of Е-cadherin was marked in 28,33% of patients with ductal adenocarcinoma, it was significantly more expressed in the areas of tumour invasion into the wall of duodenum.Conclusion. In patients with ductal adenocarcinoma direct correlation of medium strength between the levels of EGFR and HER-2/neu, EGFR and Кі-67, Кі-67 and HER-2/neu expression by the tumour cells is marked, inverse strong correlation between the level of EGFR and Е-cadherin expression, inverse correlation of medium strength between the level of expression of HER-2/neu and Е-cadherin, and between the level of Кі-67 and Е-cadherin expression there is weak inverse correlation.

Detection of Her-2/neu expression in gastric cancer: Quantitative PCR versus immunohistochemistry.

The aim of this study was to compare quantitative polymerase chain reaction (qPCR) with immunohistochemistry (IHC) for the detection of Her-2 in gastric cancer, and to investigate the correlation between the expression levels of human epidermal growth factor receptor 2 (Her-2) and clinical features. Clinical data from 426 cases of gastric cancer were collected. Her-2 expression levels in cancerous tissue were detected using IHC, and the Her-2/neu gene expression levels were determined by qPCR. The correlation between the expression level of Her-2 and clinical features was investigated. The positive expression rate of Her-2 in cancerous tissue detected using qPCR and IHC was 11.17% (46/412) and 13.38% (57/426), respectively. The positive expression of the Her-2 protein/gene was significantly correlated with the depth of invasion and lymphatic metastasis, as well as the TNM stage (P<0.05). No significant correlation was identified between positive expression of the Her-2 protein/gene and tumor location, age, gender, differentiation degree and Lauren classification (P>0.05). The diagnostic consistency was good between the two methods (κ=0.828). The results indicate that the expression of Her-2/neu is closely associated with the development of gastric cancer. qPCR is a convenient, objective and efficient method, which may be used as an alternative to IHC or fluorescence in situ hybridization for the detection of Her-2/neu gene.

Overexpression of HER-2/neu in patients with prostatic adenocarcinoma.

Prostatic adenocarcinoma is one of the main causes of cancer death, and its timely diagnosis and preventing its progression dramatically helps improve life indexes. Given the high disease recurrence rate, today, research is more inclined toward exploring causes of recurrence and development, and innovation of modern treatment methods. Several studies have explored over-expression of human epidermal growth factor receptor 2 (HER-2/neu) in prostatic cancer so far, with different results. Thus, it was decided to investigate HER-2/neu overexpression in patients with prostatic adenocarcinoma in Iran. A sample size of 40 patients with prostate cancer entered the study, using a cross-sectional, non-randomized sampling method. Parameters studied included patient age at surgery, Gleason score, serum prostatic specific antigen (PSA) before surgery, and positive sample rate after immunohistochemical staining to investigate HER-2/neu overexpression. In terms of HER-2/neu receptor staining rate, of 40 slides, 16 (40%) scored 0, 13 (32.5%) 1+, 7 (17.5%) 2+, and 4 (10%) 3+. In total 27.5% of slides showed HER-2/neu overexpression. In terms of age, an inverse correlation was found (-0.181), but without significance (p=0.263). In terms of serum PSA, the correlation coefficient was 0.449 (p=0.004). With respect to Gleason score, the coefficient was 0.190 (p=0.240). In this study, HER-2/neu overexpression occurred in 27.5% of prostate cancer cases, which is a relatively high figure, compared to similar studies elsewhere. While, we failed to reveal any relationship between HER-2/neu expression status with progression and prognosis of disease, it was demonstrated that the serum PSA level was significantly higher in cases with increased receptor expression.

HER-2/neu is an independent prognostic factor in type I endometrial adenocarcinoma.

To define the prognostic significance of HER-2/neu and PTEN expression in patients with endometrioid (type I) endometrial cancer. Seventy-seven patients with endometrioid endometrial carcinoma were included in the study, in a period between 1996 and 2009. Patients with coexisting malignancy and those having incomplete immunohistochemical data or clinical follow-up were excluded. Histological staging was defined according to the revised FIGO staging (2009). Clinico-pathologic and immunohistochemical characteristics were correlated in a multivariate Cox regression analysis with overall survival (OS), cancer-related survival (CRS) and disease-free survival (DFS). Mean age of the patients was 62.7 years. The median follow-up was 67 months (9-124 months). HER-2/neu expression was detected in 18.2 % (n = 14), and PTEN expression in 72.7 % (n = 56) of our patients. Multivariate Cox regression analysis showed that patient's age, FIGO staging and HER-2/neu expression were independent prognostic factors for OS, CRS and DFS. PTEN expression did not significantly affect survival outcomes of the present study. HER-2/neu but not PTEN expression is an independent prognostic factor for type I endometrial carcinoma.

Correlation of Breast Cancer Subtypes Based on ER, PR and HER2 Expression with Axillary Lymph Node Status

Background:Lymph node metastasis is the most important prognostic factor in breast cancer patients. The present study was carried out to evaluate the association between ER, PR and HER-2/neu expression status, separately and in combinations with axillary lymph node involvement. Patients and methods: Two hundred and fifty eight breast cancer patients treated with modified radical mastectomy with axillary lymph node dissection were included. The primary tumor tissue and axillary lymph node were evaluated histologically and the expression of ER, PR and HER-2/neu was evaluated by immune his to chemistry. Results: ER and PR expression were demonstrated in 78.7% and 76.4%, respectively and over-expression of HER-2/neu was detected in 13.2% of cases. Triple positive breast cancer is more likely to have axillary lymph node metastasis and ER+/PR+/HER-2- (PPN) is the most protected group (p<=0.001). There was a strong correlation between tumor size and tumor grade with lymph node involvement (p= 0.0001 and 0.024, respectively). ; Conclusion: triple positive breast carcinomas are more likely axillary lymph node metastasis. Tumor size, tumor grade and pathological subtypes correlate with axillary lymph node status. Further confirmatory studies with a larger number of patients are necessary to define factors predicting axillary lymph node status.

English

Breast carcinoma is the most common cancer in women and leading cause of cancer death in women. The role of hormone receptor status and Her-2/neu expression as a prognostic and therapeutic tool in breast carcinoma is widely accepted. The present study was undertaken to observe the trend and to analyze the status of hormone receptors and Her-2/neu expression in breast carcinoma at our hospital and its correlation with tumor grading.

Regional differences in breast cancer biomarkers in american Indian and Alaska native women.

Breast cancer is not a homogeneous disease, but several different and unique subtypes defined by gene expression analysis. Incidence and mortality rates vary by almost 3-fold between Alaska (highest) and the Southwestern tribes (lowest). We hypothesized that these differences may be due to, in part, varying levels of biologic tumor aggressiveness. A biorepository of the North Central Cancer Treatment Group with 95 cases of American Indian and Alaska Native (AIAN) women with adenocarcinoma of the breast surgically treated from 1990 to 2000 was tested for several biomarkers. Comparison distributions of biomarker values across state of residence using t tests for continuous (p53, MIB-1, cyclin D) and ordinally scaled markers [EGF receptor (EGFR), BCL-2, Her2] and χ(2) tests of significance for binary markers [estrogen receptor (ER), progesterone receptor (PR)] were done. Significant regional differences in some biomarker expression levels were seen. No increase was observed in "triple-negative" breast cancer or Her2 overexpression in these cases. Despite a 3-fold difference in breast cancer mortality in Alaska Native versus Southwestern American Indians, standard biomarkers such as ER, PR, and Her2 neu expression did not explain the disparity. There is a need for research to understand the biologic basis of breast cancer disparities in AIAN women. Potential for a prospective trial will be explored with tribes.

Inhibition of cell growth by BrMC through inactivation of Akt in HER-2/neu-overexpressing breast cancer cells.

We previously reported that chrysin (ChR) and its analogs induced cell cycle arrest and apoptosis in human estrogen receptor-positive/-negative breast cancer cells. However, it was unknown whether 8-bromo-7-methoxychrysin (BrMC), a novel synthetic ChR analog, inhibited the cell growth of human epidermal growth factor receptor 2 (HER-2)/neu-overexpressing breast cancers. In the present study, it was demonstrated that BrMC preferentially inhibited the cell viability of HER-2/neu-overexpressing MDA-MB-453 and BT-474 cells. Western blot analysis revealed that HER-2/neu expression and tyrosine phosphorylation were inhibited by BrMC in a concentration-dependent manner; whereas the proteasome inhibitor, MG-132, significantly prevented BrMC-induced HER-2/neu depletion and cell death in MDA-MB-453 cells. This directly indicated that BrMC-induced HER-2/neu depletion and cell growth inhibition was mediated by a proteasomal pathway. BrMC significantly downregulated the expression of cyclin D1, cyclin E and CDK4, followed by the suppression of protein kinase B phosphorylation and downstream effectors, GSK-3β and β-catenin. A colony formation assay also confirmed the growth-inhibitory effects of BrMC. Thus, these findings clearly demonstrate the anticancer activity of BrMC against human HER-2/neu-overexpressing breast cancer cells. Thus, these findings clearly demonstrate the anticancer activity of BrMC against human HER 2/neu-overexpressing breast cancer cells, and highlight BrMC as a promising candidate for breast cancer therapy.

Correlation between HER-2/neu(erbB-2) expression level and therapeutic effect of combination treatment with HERCEPTIN and chemotherapeutic agents in gastric cancer cell lines

IntroductionAlthough advanced gastric cancer has many limitations and response rate is marginal in chemotherapy. Overexpression of human epidermal growth factor receptor 2(HER-2/neu) gene and its protein are associated with increased cell division and a high rate of tumor growth and have been reported in several malignancies. Especially, approximately 30% of breast cancer patients have overexpression of HER-2/neu protein and the overexpression metastasize faster, induces resistance of the chemotherapy and down-regulate function of estrogen receptor. Recombinant humanized anti-HER2 antibody (Herceptin) inhibits proliferation of HER-2/neu overexpressing tumor cells and the use of that in combination in metastatic breast cancer have increased cytotoxicity of chemotherapeutic agents.MethodsWe evaluated the expression of HER-2/neu protein in gastric cell lines by FACS and then comparing the cytotoxicity in chemotherapeutics (doxorubicin, cisplatin, paclitaxel, 5-FU) alone and in combination with Herceptin according to the expression of HER-2/neu protein by MTT assay.Results1. NCI-N87 (88%) gastric cancer cell line and SK-BR-3 (89%) breast cancer cell line with strong positivity of HER-2/neu expression. YBC-2 (55%) and YBC-3 (48%) gastric cancer cell line with intermediated, weak positivity respectively. Negative control U-87 MG (6%) brain cancer cell line were showed low expression of HER-2/neu. 2. Cell growth was dose-dependently inhibited in HER-2/neu positive, control cell line SK-BR-3 by Herceptin treatment but not observed in HER-2/neu negative control cell line U-87 MG. Effective growth inhibition was not observed in gastric cancer cell lines with single treatment of Herceptin, all cell lines observed the dose-dependent growth inhibition to chemotherapeutic agents (doxorubicin, cisplatin, paclitaxel and 5-FU). 3. Combination of Herceptin with doxorubicin observed synergistic effects in all cancer cell lines except YBC-3, combination of Herceptin with cisplatin observed NCI-N87 and SK-BR-3 and combination of Herceptin with paclitaxel observed synergistic effects in YBC-2. Combination of Herceptin with 5-FU observed antagonistic effects in all cancer cell lines.ConclusionsAccording to HER-2/neu expression level, effect of anti-cancer agents was observed differently in combination of Herceptin with chemotherapeutic agents. This suggests that HER-2/neu expression level can be applied standard of combination drug selection in combination of Herceptin With chemotherapeutic agents in gastric cancer.

Impact of the nottingham prognostic index (NPI) on the occurrence of relapses compared with the HER-2/neu expression in breast cancer patients

Introduction Aim of this study was to determine the influence of clinical, histopathological and immunohistochemical parameters and the Nottingham prognostic index (NPI) on the occurrence of relapses compared with the HER-2/neu overexpression in breast cancer patients. Materials and methods In this study, 174 patients with primary breast cancer were analyzed in the period from June 2007 to June 2010. Clinical and histopathological parameters with the NPI, and additional immunohistochemical parameters were determined (HER-2/neu status, Ki67 and p53 protein product). During the follow-up period (42–80 months) relapses were observed in 38 patients (21.8%). Results The age of patients ranged from 28–83 years (mean 55.48+10.0). Positive lymph nodes were observed in 99 (57%) patients and in 70 (40%) patients; the tumor was poorly differentiated. HER-2/neu overexpression was observed in 33 (19%) patients, and in 13 (40%) of them, relapses were present. NPI was higher than 3.4 in 90% (30/33, HER2/neu positive patients) while HER-2/neu overexpression was associated with the tumor size, lymph-nodal status, mitotic index, degree of histological differentiation, lymph-vascular invasion, stage of the disease, Ki67, and p53 expression (p < 0.05). Conclusion NPI score is an excellent parameter to evaluate the course of disease in HER-2/neu positive breast cancer compared to occurrence of relapses. Aim of this study was to determine the influence of clinical, histopathological and immunohistochemical parameters and the Nottingham prognostic index (NPI) on the occurrence of relapses compared with the HER-2/neu overexpression in breast cancer patients. In this study, 174 patients with primary breast cancer were analyzed in the period from June 2007 to June 2010. Clinical and histopathological parameters with the NPI, and additional immunohistochemical parameters were determined (HER-2/neu status, Ki67 and p53 protein product). During the follow-up period (42–80 months) relapses were observed in 38 patients (21.8%). The age of patients ranged from 28–83 years (mean 55.48+10.0). Positive lymph nodes were observed in 99 (57%) patients and in 70 (40%) patients; the tumor was poorly differentiated. HER-2/neu overexpression was observed in 33 (19%) patients, and in 13 (40%) of them, relapses were present. NPI was higher than 3.4 in 90% (30/33, HER2/neu positive patients) while HER-2/neu overexpression was associated with the tumor size, lymph-nodal status, mitotic index, degree of histological differentiation, lymph-vascular invasion, stage of the disease, Ki67, and p53 expression (p < 0.05). NPI score is an excellent parameter to evaluate the course of disease in HER-2/neu positive breast cancer compared to occurrence of relapses.

Assessment of Her‐2/neu status using immunocytochemistry and fluorescence in situ hybridization on fine‐needle aspiration cytology smears: Experience from a tertiary care centre in India

Breast carcinoma shows amplification/overexpression of Her-2/neu in ∼20-30% of cases. The determination of Her-2/neu expression accurately is vital in clinical practice as it has significant predictive value and eligibility for anti Her-2/neu therapy. Amplification and overexpression of Her-2/neu gene is traditionally identified by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) on tissue sections; only a few studies have evaluated feasibility of these techniques on cytological smears. One hundred cases of breast cancer with fine-needle aspiration cytology (FNAC) samples and corresponding surgically resected specimen were selected. Immunocytochemistry (ICC) and FISH for Her-2/neu was done on FNA smears, whereas IHC was performed on corresponding tissue sections. Diagnostic accuracy of ICC was 99% when compared with IHC. Comparison of FISH results with IHC showed 100% concordance. Unlike many centers in West, FNAC is still routinely performed in developing countries like India where vast majority of breast cancer cases present as palpable lumps. The high rates of accuracy of ICC and FISH for Her-2/neu detection can make FNAC a relevant first line of investigation as a cost effective model with a rapid turn-around time, providing complete information necessary for initial management of breast cancer patients.

Abstract P2-09-07: Inhibition of aurora-A by MLN8237 decreases SMAD5 expression and increases effectiveness of chemotherapeutic agents in breast cancer cells

Abstract BACKGROUND: Breast cancer affects thousands of women each year. Epithelial to Mesenchymal transition (EMT) has been associated with increased metastatic potential of cancer cells as well as resistance to chemotherapy. Predicatively, presence of EMT leads to worse prognosis in patients with breast cancer. We have recently showed that Aurora-A plays a key role in development of EMT and increased ability of breast cancer cells for self renewal. Therefore we hypothesized that inhibition of Aurora-A in breast cancer cells will lead to increased sensitivity to chemotherapy. RESULTS: Increased Aurora-A expression, as assessed by IHC staining, in patients with locally advanced disease was associated with worse prognosis. We show that in ER+ cell line, MCF7, activating MAPK pathway through overexpression of Raf1 leads to increase in Aurora-A activity. These cells have decreased sensitivity to treatment with paclitaxel when compared to untransfected cells in MTT assays. This effect is even more exacerbated with over-expression of Aurora-A. The sensitivity to paclitaxel was restored with inhibition of Aurora-A by specific inhibitor, Alisertib. This was further explored in triple negative (MDA-MB-231) or Her-2/neu expressing (SUM149) cell lines. These cells express higher levels of Aurora-A. Inhibition of Aurora-A activity in combination with paclitaxel was superior when compared to either therapy alone. Similar effect was seen with use of anthracyclines. Inhibition of Aurora-A resulted in decreased SMAD5 expression as well as decreased Akt phosphorylation. Current studies are looking at a role of Aurora-A in activating SMAD5 and its role in resistance to chemotherapy. In vivo experiments evaluating combination therapies in breast cancer animal model are ongoing. CONCLUSIONS: Inhibition of Aurora-A by a specific inhibitor, MLN8237, resulted in decreased SMAD5 expression and increased effectiveness of chemotherapeutic agents in breast cancer cells. These results contribute to better understanding of signaling pathways involved in resistance of breast cancer cells to chemotherapy. This knowledge could be very useful in developing more effective treatments for breast cancer patients in neo-adjuvant, adjuvant and metastatic settings. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-09-07.

Abstract P2-18-11: A multi-center prospective study of image-guided radiofrequency ablation for small breast carcinomas

Abstract Background: As the management of breast carcinoma evolves toward less invasive treatments, the next step is the possibility of removing the primary tumor without surgery. The most promising noninvasive ablation technique is radiofrequency ablation (RFA), which can effectively kill tumor cells with a low complication rate. Our preliminary studies of RFA followed by standard surgical resection have indicated that this technique is effective for surgical ablation of small (≤ 2cm) breast tumors without extensive intraductal components (EIC). Methods: To determine if RFA is oncologically and cosmetically appropriate for the local treatment of primary breast carcinoma, this multi-center prospective study used RFA as the sole local treatment of breast tumors ≤ 1.5cm in size on ultrasound and MRI. Exclusion criteria include receiving of preoperative chemotherapy, or the presence of invasive lobular carcinoma or invasive ductal carcinoma with suspicious EIC. After confirmation that the standard baseline core biopsy for diagnosis and measurement of tumors markers (ER, PgR, HER-2/neu expression and the presence of the Ki-67 proliferative marker) have been obtained, consent will be obtained and the patient scheduled RFA. All patients received adjuvant radiation therapy. The use and choice of systemic therapy will be based on the information from the baseline core biopsy and imaging studies. The first primary endpoints of this study is successful tumor ablation, as evidenced by negative findings on vacuum-assisted or core biopsies and imaging studies after RFA. The second primary endpoints is the incidence of procedure related adverse events. Forty patients with small tumors that are clearly identifiable and measurable by ultrasound and MRI were enrolled. The response to ablation was evaluated with both vacuum-assisted or core biopsies and imaging studies every 3 months during the first year. The long-term outcomes were assessed using quality of life measurement scales and imaging studies every 6 months thereafter through year 5. Results: Of the 58 patients who participated in this study, 55 completed the protocol. In 50 of the 55 (91%) treated patients, successful tumor ablation, as determined by negative findings on vacuum-assisted or core biopsies and imaging studies, was confirmed. In these 50patients, there were no local or distant recurrences after 1 year. The remaining 5 patients with biopsies positive for residual tumor underwent surgical resection. In 46 of the 50 patients with successful ablation, cosmetic outcomes were excellent after 1 year and only 2 patients experienced minor skin burns during the tumor ablation. No other adverse events were reported. Conclusions: RFA can be safely used alone in patients with small breast tumors, provided that local tumor control must be regularly assessed by image-guided vacuum-assisted or core biopsies after ablation. RFA in patients with early breast carcinoma can achieve local control rates similar to those with wide local excision, but with improved cosmetic results. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-18-11.

Abstract A115: Withaferin A downregulates her-2/neu protein expression in human breast cancer cells.

Abstract HER-2/neu protein is over-expressed in approximately 20-30% of breast cancers. Overexpression of this oncoprotein results in a more aggressive tumor phenotype and poor prognosis for patients. Current treatment strategies include passive administration of anti-HER-2/neu monoclonal antibody Trastuzumab as well as the small molecule inhibitor lapatinib. Both of these molecules function at the receptor level to block HER-2/neu signaling resulting in inhibition of cell growth and tumor regression in the majority of patients. However, a significant number of patients relapse or develop progressive disease due to acquired mechanisms of resistance to current therapy. Alternative approaches, that utilize multiple modes of action, are therefore necessary to combat this severe form of breast cancer. Withaferin A (WA) is a natural compound isolated from the Indian ayurvedic medicinal plant Withania Somnifera. Natural compounds are notorious for their multiple mechanisms of action, which make them ideal therapeutics for complex diseases such as cancer. Recent research has demonstrated that WA is effective in inducing cell death in several cancer cell lines by inhibiting molecular pathways such as Akt and MAPK proliferative cascades which are downstream of her-2/neu activation. Additionally, WA has also been shown to inhibit function of the molecular chaperone heat shock protein 90 (HSP90) in pancreatic carcinoma cells. Since HER-2/neu is a client protein of HSP90 and the chaperone functions to stabilize HER-2/neu protein expression on the cell surface, our goal was to determine whether WA would directly affect HER-2/neu expression. We initially established the cytotoxic activity of WA on HER-2/neu expressing human breast cancer cell lines such as SKBr3, MCF-7her-2neu and BT474 by MTT assay. We report that WA exhibits potent cytotoxic activity in these cells with IC50 concentrations of 1.6, 2.1 and 0.6 μM respectively after 48hr treatment. Comparison of WA sensitivity in a low HER-2/neu expressing cell line MCF-7 generated an IC50 of 5.1μM, indicating that cell lines that overexpress HER-2/neu may be more susceptible to the compound. Analysis of HER-2/neu expression both at the cell surface and intracellular localization were next performed in response to WA. WA treatment completely downregulated the expression of HER-2/neu protein from cell surface by 24hr and was both time and concentration dependent. Abrogation of intracellular protein expression also correlated to the downregulation from the cell surface. We determined that WA does not alter transcription of HER-2/neu, as there was no change in the gene expression with WA treatment compared to untreated cells. Further experiments are ongoing to address whether WA specifically targets signaling pathways that alter HER-2/neu signaling. Understanding the mechanisms by which WA induces down-regulation of HER-2/neu would serve as a means of developing WA as an alternative therapeutic strategy for HER-2/neu cancers not amenable to standard treatments. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A115. Citation Format: Ananlise Smith, Sarah Poliquin, Darlah Lopez-Rodriguez, Samita Andreansky. Withaferin A downregulates her-2/neu protein expression in human breast cancer cells. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A115.

Clinical instability of breast cancer markers is reflected in long-term in vitro estrogen deprivation studies.

BackgroundLong-term estrogen deprivation models are widely employed in an in vitro setting to recapitulate the hormonal milieu of breast cancer patients treated with endocrine therapy. Despite the wealth information we have garnered from these models thus far, a comprehensive time-course analysis of the estrogen (ER), progesterone (PR), and human epidermal growth factor 2 (HER-2/neu) receptors on the gene and protein level, coupled with expression array data is currently lacking. We aimed to address this knowledge gap in order to enhance our understanding of endocrine therapy resistance in breast cancer patients.MethodsER positive MCF7 and BT474 breast cancer cells were grown in estrogen depleted medium for 10 months with the ER negative MDA-MB-231 cell line employed as control. ER, PR and HER-2/neu expression were analysed at defined short and long-term time points by immunocytochemistry (ICC), and quantitative real-time RT-PCR (qRT-PCR). Microarray analysis was performed on representative samples.ResultsMCF7 cells cultured in estrogen depleted medium displayed decreasing expression of ER up to 8 weeks, which was then re-expressed at 10 months. PR was also down-regulated at early time points and remained so for the duration of the study. BT474 cells generally displayed no changes in ER during the first 8 weeks of deprivation, however its expression was significantly decreased at 10 months. PR expression was also down-regulated early in BT474 samples and was absent at later time points. Finally, microarray data revealed that genes and cell processes down-regulated in both cell lines at 6 weeks overlapped with those down-regulated in aromatase inhibitor treated breast cancer patients.ConclusionsOur data demonstrate that expression of ER, PR, and cell metabolic/proliferative processes are unstable in response to long-term estrogen deprivation in breast cancer cell lines. These results mirror recent clinical findings and again emphasize the utility of LTED models in translational research.