Neu Gene Amplification Research Articles

Her-2/neu gene amplification and its correlation with other prognostic factors in Iranian breast cancer patients.

Breast cancer is one of the most common diseases in females, arising from overexpression of a variety of oncogenes like HER2/neu. The amplification rate of this gene is variable in different breast cancer patients. In this study, the amplification of the HER2/neu oncogene was distinguished in breast cancer patients and its correlation with prognostic factors. Also, the simultaneous effect of prognostic factors on the occurrence of a specific prognostic factor was investigated. The multiplex PCR technique was used to assay the amplification of the HER2/neu oncogene in breast cancer patients. After extracting DNA from 100 tumor tissue and 8 normal breast tissue samples, the amplification of the HER2/neu gene was distinguished by the co-amplification of a single-copy reference gene, γ-IFN, and the target gene HER2/neu in the PCR reaction and using the Gel analyzer software. SPSS 23 and STATA 9.1 software were used for statistical analysis. The HER2/neu gene was amplification in 30% of the tumor samples. The statistical analysis showed a statistically significant relationship between HER2/neu gene amplification and progesterone receptors. Amplification of the HER2/neu gene significantly increases the chance of lymph node involvement. Also, the amplification of this gene in tumors with histological grade II tissue is more than grade I. The amplification of the HER2/neu gene can be used as an independent prognostic factor in predicting lymph node involvement and histological grade in breast cancer patients.

MULTIMODALITY TESTING OF HER2/NEU MARKER IN BREAST CANCER: A CORRELATIVE STUDY

Currently HER-2/neu status in breast cancer is categorized by immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) in clinical practice. The amplification of the HER-2/neu gene has a prognostic significance. Quantitative determination of HER-2/neu gene amplification by qPCR or over-expression by qRT-PCR is not routinely used, whereas qPCR based assays can be an alternative method to evaluate HER-2/neu status. We searched literature and databases including the National Institutes of Health’s (NIH), National Library of Medicine (NLM), The National Center for Biotechnology Information (NCBI), PubMed, and Google Scholar. The current study aimed to identify the correlation between FISH, IHC, and real-time-PCR reaction in measuring HER-2/neu expression in breast cancer.

Comparison of Her-2/Neu Gene Amplification or Expression between IHC and FISH in BC.

This study aimed to analyze and compare the application of FISH and IHC in BC tissue Her-2 / neu gene amplification or expression. For this purpose, 110 patients with BC were selected. FISH and IHC were performed on the BC tissues that were surgically removed, and the results were compared and analyzed. The results showed that in 110 BC tissues, the expression of HER-2 protein was (+++) 25 cases (22.73%); (++) 44 cases (40%); (+) 26 cases (23.64%); (1) 15 cases (13.64%). There were 42 cases of HER-2 gene amplification in 110 BC tissues, and 68 cases had no amplification. IHC test positive (+++) is consistent with FISH positive coincidence rate, IHC test negative (+/-) is consistent with FISH negative coincidence rate, IHC test suspicious positive (++) compared with the FISH result, the difference is statistically significant. However, the total coincidence rate between IHC test results and FISH test results was 89.29 (25/28), and the two test methods were positively correlated. Generally, IHC positive and negative expressions are in good agreement with FISH test results. The suspected positive expression of IHC is inconsistent with the FISH test result, suggesting that the IHC test is suspicious, and the positive specimens need to be tested by FISH.

Impact of ASCO/CAP 2007, 2013 and 2018 recommendations on Her-2/neu gene amplification status testing in patients with breast cancer

Aim. To Assess the significance of changes in the diagnostics of Her-2/neu gene amplification status using the ASCO/CAP 2007, 2013 and 2018 recommendations by retrospective evaluation of breast cancer (BC) tissue test results performed by fluorescence in situ hybridization (FISH). Methods. Her-2/neu FISH dual probe test data of 797 BC specimens were re-evaluated in accordance with the requirements of the consecutive updated ASCO/CAP recommendations. Results. The proportion of cases without Her-2/neu gene amplification by the ASCO/CAP 2013 and 2018 criteria was 11,0 % lower than by the 2007 ASCO/CAP criteria. There was no statistically significant difference between the number of Her-2/neu gene amplification positive cases according to ASCO/CAP 2007, 2013 and 2018 criteria. The proportion of cases with a doubtful outcome for the use of ASCO/CAP in 2018 was significantly higher (12,2 %, p < 0,01) compared to that by ASCO/CAP 2007 and 2013 (6,6 % and 12,2 % accordingly). Conclusions. The findings of a re-benchmarking review (in accordance with the 2018 ASCO/CAP criteria) of the FISH case study results show practical differences from the previous ASCO/CAP 2007, 2013 recommendations. The proportion of samples, which needs the additional research, increases with each revision of the recommendations against the backdrop of the expansion of the case group with questionable amplification status of the Her-2/neu gene by the FISH double probe method. Keywords: ASCO/CAP, amplification status of the gene Her-2/neu, breast cancer.

HER-2 status of circulating tumor cells in a metastatic breast cancer cohort: A comparative study on characterization techniques.

BackgroundPersonalized targeted treatment in metastatic breast cancer relies on accurate assessment of molecular aberrations, e.g. overexpression of Human Epidermal growth factor Receptor 2 (HER-2). Molecular interrogation of circulating tumor cells (CTCs) can provide an attractive alternative for real-time biomarker assessment. However, implementation of CellSearch-based HER-2 analysis has been limited. Immunofluorescent (IF) image interpretation is crucial, as different HER-2 categories have been described. Major questions in CTC research are how these IF categories reflect gene expression and amplification, and if we should consider ‘medium’ HER-2 expressing CTCs for patient selection.MethodsTumor cells from spiked cell lines (n = 8) and CTCs (n = 116 samples) of 85 metastatic breast cancer patients were enriched using CellSearch. Comparative analysis of HER-2 expression by IF imaging (ACCEPT, DEPArray, and visual scoring) with qRT-PCR and HER-2/neu FISH was performed.ResultsAutomated IF HER-2-profiling by DEPArray and ACCEPT delivered comparable results. There was a 98% agreement between 17 trained observers (visual scoring) and ACCEPT considering HER-2neg and HER-2high expressing CTCs. However, 89% of HER-2med expressing CTCs by ACCEPT were scored negative by observers. HER-2high expressing tumor cells demonstrated HER-2/neu gene amplification, whereas HER-2neg and HER-2med expressing tumor cells and CTCs by ACCEPT were copy-number neutral. All patients with HER-2-positive archival tumors had ≥1 HER-2high expressing CTCs, while 80% of HER-2-negative patients did not. High relative gene expression of HER-2 measured on enriched CTC lysates correlated with having ≥1 HER-2high expressing CTCs.ConclusionAutomated images analysis has enormous potential for clinical implementation. HER-2 characterization and clinical trial design should be focused on HER-2high expressing CTCs.

HER-2/Neu and Hormone Receptor Analysis in Breast Carcinomas and Their Association with Clinicopathologic Parameters.

Invasive breast carcinomas of no special type (IC-NST) are the heterogeneous tumours showing distinct prognostic features even in patients with similar clinicopathological characteristics. To date, many clinicopathological data have been analyzed to make a guess about prognosis and to determine treatment modality. In this study, HER-2/neu status was analyzed by using both immunohistochemical (IHC) and fluorescence in situ hybridization (FISH) methods, and its correlations with hormone receptor status and clinicopathological parameters were investigated. The study was included 112 female patients with diagnosis of IC-NST. FISH for HER-2/neu was applied in only primary tumour tissues, while IHC analyses for HER-2/neu, estrogen (ER) and progesterone receptors (PR) were applied on both primary and metastatic lymph node foci. The results were compared with appropriate statistical methods. Our rates of HER-2/neu overexpression and gene amplification in the overall study group were 22.3 and 25%, respectively. In the metastatic group, these rates were higher than those of the overall study group (34% and 40%, respectively). Gene amplification rate of the axilla positive group was 40%, while this rate in non-metastatic group was 6.7% (p=0.015). Overexpression and amplification results were compliant (χ2=77,591, p<0.001). The concordance rates in HER-2/neu negative and overexpression groups were 95.3% and 88%, respectively. Our false negativity rate was 4.7%. While 36% of score 3+ cases were ER positive, 67.1% of HER-2/neu negative cases showed ER positivity (p=0.01). The increase of gene amplification rate in ER negative cases over 50 years age was more than two times and statistically significant (p=0.014). The concordance rates between the results of IHC and FISH in the HER-2 negative and the overexpression categories were compatible with the literature and lower than the literature, respectively. In the case of ER negativity, the patient's age over 50 years was associated with a higher rate of gene amplification.

Assessment of human epidermal growth factor receptor 2/neu gene amplification and expression as a biomarker for radiotherapy and hormonal-treated breast cancer patients in upper Egypt.

Breast cancer plays major public health in Egyptian women. In upper Egypt, There is an increase in the incidence of breast cancer compared to other Egyptian areas without know the reasons. In this study, we aimed to evaluate the potential of HER-2/neu status as one of the important markers to classify the women suffering from breast cancer in upper Egypt and monitoring the responsiveness to different therapies. The present study was performed on 67 female breast cancer patients in the South Egypt Cancer Institute to evaluate HER-2/neu gene amplification and expression. Tissue samples were used for immunohistological analysis of endocrine receptors, HER-2/neu, and HER-2/neu gene amplification. In addition, the blood samples were also used to determine HER-2/neu gene expression. All statistical analyses were performed using Chi-square test. The statistical difference is considered statistically significant at P < 0.05. There was a statistically significant association between HER-2/neu gene expression and the age of patients. There is decrease in the level of HER-2/neu mRNA expression in group treated with chemotherapy and group treated with chemotherapy and radiotherapy compared to each group baseline level of HER-2/neu mRNA expression before treatment. On the contrary, the group treated with chemotherapy, radiotherapy, and hormonal therapy revealed increase on the level of HER-2/neu mRNA expression when compared with their baseline for the same patients before treatment. We need further studies on the large group of upper Egypt breast cancer patients to confirm that the level of HER-2/neu mRNA expression can be used as a marker for classified them and their response to different treatment.

HER-2/neu alterations in non-small cell lung cancer: A comprehensive evaluation by fluorescence in situ hybridization and immunohistochemistry

Aim: The molecular relationships of protein overexpression and HER-2/neu gene amplification in lung cancers are not clearly defined. The aim of the study is to evaluate HER-2/neu changes in non-small cell lung cancer with in situ hybridization and immunohistochemistry, to compare the effectiveness of both methods and to examine the influence of HER-2/neu gene and protein level alterations on prognosis. Material and Methods: HER-2/neu alterations were analyzed with fluorescence in situ hybridization and immunohistochemistry in 24 patients with non-small cell lung cancer. Results: Of 24 patients, 13 (54%) were found immunohistochemistry (IHC) positive, on the other hand 24 (100%) were found fluorescence in situ hybridization (FISH) negative. Balanced trisomy was detected in 23 out of FISH negative cases and balanced quadrisomy was detected in one case. Our study has revealed that overexpression of HER-2/neu protein is not associated with real gene amplification. No significant correlation was detected between HER-2/neu overexpression with amplification, and prognosis of non-small cell lung cancer. Conclusion: No correlation is present between fluorescence in situ hybridization and immunohistochemistry results. HER-2/neu alterations are present in non-small cell lung cancers. Therefore, HER-2/neu overexpression may be associated with transcriptional mechanisms rather than gene amplification.

Assessment of HER-2/neu, с-MYC andCCNE1 gene copy number variations andprotein expression in endometrial carcinomas.

To analyze copy number variations of HER-2/neu, c-MYC and CCNE1oncogenes and their protein expression in endometrioid endometrial carcinomas in relation to the degree of tumor progression and presence of a family history of cancer in cancer patients. The study was conducted on endometrial cancer (EC) samples from 68patients with I-II FIGO stages of disease. Copy number analysis of HER-2/neu, c-MYC and CCNE1genes was performed by quantitative PCR. Protein expression was analyzed using immunohistochemistry. Assessment of copy number variations of HER-2/neu, c-MYC and CCNE1genes revealed their amplification in the tumors of 18.8, 25.0and 14.3% of EC patients, respectively. High expression of corresponding proteins was detected in 14.6, 23.5and 65.6% of patients, respectively. It was established that HER-2/neu gene amplification is more common in the group of tumors of low differentiation grade than in moderate grade EC (35.7and 5.5% of cases, respectively, p < 0.05). Also, high expression of c-Myc protein was more frequently observed in low differentiated tumors compared to the moderately differentiated EC (36.6and 13.2% of cases, respectively, p < 0.05). Expression of HER-2/neu and cyclin E proteins was found to be dependent on the depth of tumor invasion into the myometrium. High expression of HER-2/neu protein was observed in 25.0and 4.1% of EC patients with tumor invasion > ½ and < ½ of the myometrium, respectively, and cyclin E- in 86.7and 46.6% of cases, respectively, p < 0.05. It was shown that among patients with a family history of cancer, a larger proportion of cases with high expression of c-Myc protein was observed compared to the group of patients with sporadic tumors (43.8and 17.3%, respectively; p < 0.05). Amplification of HER-2/neu gene, along with high expression of c-Myc, HER-2/neu and cyclin E proteins, are associated with such indices of tumor progression as a low differentiation grade and deep myometrial invasion, suggesting the potential possibility of including these markers in the panel for determining the molecular EC subtype associated with an aggressive course of the disease. In a certain category of EC patients, there is a relationship between a family history of cancer and high expression of c-Myc protein.

Her-2/neu Oncogene Amplification by Fluorescence In Situ Hybridization and Protein Overexpression on Immunohistochemistry in Breast Cancer.

To investigate the concordance and discordance between the test results of Her-2/neu by immunohisto-chemistry (IHC) and flourescence In Situ hybridization (FISH) in breast cancer cases. Descriptive cross-sectional study. Department of Histopathology, Dr. Ziauddin Hospital, Karachi, from 2011 to 2016. Forty-three specimens of invasive ductal carcinoma of breast were evaluated for grade and Her-2/neu status using IHC and FISH methods. Concordance and discordance between their results was determined. There is 100% concordance between FISH and IHC in cases scoring 0, 1+ (negative) and 3+ (positive) immunostaining. Tumour cases scoring 2+ immunostaining showed amplification in 69.2% cases. All grade-I tumours were non-amplified on FISH, while most of the grade-III tumours showed Her-2/neu amplification on FISH. There is significant association of Her-2/neu IHC with tumour grade and FISH (p<0.05). A fairly high proportion i.e. 69.7% of cases showed Her-2/neu gene amplification. There was high concordance between Her-2/neu testing on IHC and FISH, (Kappa co-efficient 0.466, p <0.001). Her-2/neu amplification increases with increasing grade of breast cancer. A high proportion of Her-2/neu gene amplified cases indicates aggressive disease in that area and need for FISH testing on large scale, which is the gold standard for equivocal cases on immunohistochemistry.

Determination of HER-2/neu gene Status by Chromogenic in Situ Hybridisation Assay on Borderline (2+) Immunohistochemistry Cases in Patients with Invasive Breast Carcinoma: An Experimental Study on Preserved Tissue

HER-2 (also known as HER-2/neu) is a member of the epidermal growth factor (EGF) receptor family. The amplification of oncogene HER-2 is presented in 20 to 30% of breast cancers and results in an increase of the protein expression. HER-2 over expression has also been shown to correlate with poor prognosis. It is associated with poorly differentiated high grade tumors with lymph node involvement, greater risk of recurrence and relative resistance to some types of chemotherapy. The receptor is however a target for treatment with anti HER-2 antibody trastuzumab (Herceptin). Immunohistochemistry analysis for HER 2 scoring is subjective, requires trained personnel and expertise. One of the main concerns with IHC is that there is evidence of significant inter-observer variation in the assessment of staining, which can lead to misclassification of HER2 status. Chromogenic in situ hybridization (CISH) testing is sensitive and specific in detecting HER-2/neu gene amplification. Direct evaluation of gene amplification using CISH assay is a reliable method for routine diagnostic evaluation of HER2/neu status in breast cancer patients, especially in specimens showing 2+ IHC score. This observational experimental study was carried out in the Department of Pathology, Bangabandhu Sheikh Mujib Medical University, Dhaka, during the period of July 2014 to June 2015, The aim of this study was to assess HER-2 expression accurately in equivocal immunohistochemistry (2+) invasive breast cancer cases by Chromogenic in situ hybridization (CISH) and to associate the findings of CISH assay with the histologic prognostic and predictive factors (eg: tumor size, regional lymph node metastases, tumor grade and type. A total of 20 archival paraffin tissue blocks and IHC slides with IHC score 2+ for HER-2 were included in this study. All the slides were reviewed. CISH assay was done on section from archival paraffin block.CISH assay showed amplification of the HER-2 neu gene in 30% of cases. Majority (60%) was nonamplified. In two cases the results were unsatisfactory for interpretation. The differences were not statistically significant (p&gt;0.05) among three groups (amplified, nonamplified and unsatisfactory) regarding the baseline characteristics (age and sex) with CISH. All the results of CISH assay in association with tumor size, tumor grade and lymphnode metastasis were not statistically significant. HER-2 / neu status is the one of the most important prognostic and predictive factor. It is a target for treatment with anti HER-2 antibody trastuzumab (Herceptin). Detection of HER-2/ neu status by IHC may sometimes be difficult and inaccurate, specially in IHC score 2+ cases. Based on the findings of the study, CISH can be considered a useful, simple and reproducible method for detecting HER-2/ neu gene amplification in cases with borderline (2+) immunohistochemistry score , and patients may be benefited from Trastuzumab therapy.

HER-2/neu gene amplification in gastric adenocarcinoma and its relationship with clinical and pathological findings.

Amplification of the HER-2/neu oncogene influences the progression of gastric cancer, its prognosis, and therapy. A precise examination of HER-2/neu-amplified tumor tissue is essential for managing disease and prescribing the appropriate treatment. This study aimed to investigate the status of HER-2/neu gene in the gastric cancer samples and its relationship with clinical and pathological information. In this study on 80 paraffin-embedded tissue samples from patients with gastric adenocarcinoma [2006-2011], DNA was extracted to quantify the gene expression levels of HER-2 using a polymerase chain reaction (PCR) method. Data were statistically analyzed by chi-square test using SPSS16.0 software. PCR results indicated that HER-2/neu gene amplifications occurred in 58 of the 80 samples (72.5%). HER-2/neu gene expression was not significantly related to age and sex, but the larger tumor size and the more advanced stage were significantly associated with HER-2/neu overexpression. The data show the HER-2/neu gene is more amplified in stage 4 of gastric cancer with a larger size of mass. Older age and male sex also appear to be more associated with HER-2/neu gene expression.

Recent Advances in Comprehending the Signaling Pathways Involved in the Progression of Breast Cancer.

This review describes recent advances in the comprehension of signaling pathways involved in breast cancer progression. Calcium sensing receptor (CaSR), caveolae signaling, signaling referred to hypoxia-inducing factors and disturbances in the apoptotic machinery are related to more general biological mechanisms and are considered first. The others refer to signaling pathways of more specific biological mechanisms, namely the heparin/heparin-sulfate interactome, over-expression of miRNA-378a-5p, restriction of luminal and basal epithelial cells, fatty-acid synthesis, molecular pathways related to epithelial to mesenchimal transition (EMT), HER-2/neu gene amplification and protein expression, and the expression of other members of the epithelial growth factor receptor family. This progress in basic research is fundamental to foster the ongoing efforts that use the new genotyping technologies, and aim at defining new prognostic and predictive biomarkers for a better personalized management of breast cancer disease.

Enrichment and Molecular Analysis of Breast Cancer Disseminated Tumor Cells from Bone Marrow Using Microfiltration

PurposeMolecular characterization of disseminated tumor cells (DTCs) in the bone marrow (BM) of breast cancer (BC) patients has been hindered by their rarity. To enrich for these cells using an antigen-independent methodology, we have evaluated a size-based microfiltration device in combination with several downstream biomarker assays.MethodsBM aspirates were collected from healthy volunteers or BC patients. Healthy BM was mixed with a specified number of BC cells to calculate recovery and fold enrichment by microfiltration. Specimens were pre-filtered using a 70 μm mesh sieve and the effluent filtered through CellSieve microfilters. Captured cells were analyzed by immunocytochemistry (ICC), FISH for HER-2/neu gene amplification status, and RNA in situ hybridization (RISH). Cells eluted from the filter were used for RNA isolation and subsequent qRT-PCR analysis for DTC biomarker gene expression.ResultsFiltering an average of 14×106 nucleated BM cells yielded approximately 17–21×103 residual BM cells. In the BC cell spiking experiments, an average of 87% (range 84–92%) of tumor cells were recovered with approximately 170- to 400-fold enrichment. Captured BC cells from patients co-stained for cytokeratin and EpCAM, but not CD45 by ICC. RNA yields from 4 ml of patient BM after filtration averaged 135ng per 10 million BM cells filtered with an average RNA Integrity Number (RIN) of 5.3. DTC-associated gene expression was detected by both qRT-PCR and RISH in filtered spiked or BC patient specimens but, not in control filtered normal BM.ConclusionsWe have tested a microfiltration technique for enrichment of BM DTCs. DTC capture efficiency was shown to range from 84.3% to 92.1% with up to 400-fold enrichment using model BC cell lines. In patients, recovered DTCs can be identified and distinguished from normal BM cells using multiple antibody-, DNA-, and RNA-based biomarker assays.

Assessment of HER-2/neu gene amplification and expression as a biomarker for radiotherapy and hormonal treated breast cancer patients in Upper Egypt

Assessment of HER-2/neu gene amplification and expression as a biomarker for radiotherapy and hormonal treated breast cancer patients in Upper Egypt

A Study of Evaluation of Immunoreactivity of c-erbB-2 and Histopathological Parameter in Esophageal Carcinoma

Abstrct Background: Squamous cell carcinoma is commonest histologic variant of carcinoma of esophagus (ESCC). It is predominant in developing countries and usually has a poor prognosis, because of its advanced stage at the time of diagnosis. Understanding the prognostic factors helps in selection of the most appropriate surgical approach in order to select patient for target directed therapies. HER-2/neu (C-erbB-2), member of human epidermal growth factor receptors (EGFR) is important, as it plays a crucial role in therapeutic implications of esophageal carcinoma. Considering the importance of c-erbB-2 expression and its prognostic significance in esophageal cancer, this study was undertaken. Result: Fifty cases of esophageal carcinoma including squamous cell carcinoma and adenocarcinoma studied showed M: F ratio of 1.9:1 with age ranging from 32 years to 80 years (mean age 59.1 years). There were 43(86%) cases of Squamous cell carcinoma with 65.1% positivity of which 10 (20%) cases were poorly diffentiated with a positivity of 60% ,20 (40%) cases were moderately diffentiated with a positivity of 65% and 09 (18%) were well differentiated with a positivity of 55.6% for c-erbB-2 staining. Seven (14%) cases of adenocarcinoma with a positivity of 85.7% and four (08%) cases of endoscopic biopsy have a positivity of 75% for c-erbB-2 staining. Conclusion: Present study results illustrate that HER-2/neu(c-erbB-2) gene amplification is a influential prognostic indicator in ESCC. Therefore, additional studies with more cases including supplementary techniques are compelling to verify molecular alterations involved in tumor progression, thus assisting in the advancement of modern therapies for ESCC.

Abstract 1208: Androgen receptor expressing metaplastic carcinoma cell line established from non-caucasian female patient

Abstract Background: Metaplastic carcinoma (MCa) is a rare entity of breast cancer (BCa) where tumor of epithelial origin manifests differentiation into non-glandular, mesenchymal phenotype. These tumors exhibit heterogeneity with spindle, squamous, chondroid or osseous differentiation. Majority of these tumors do not express estrogen or progesterone receptors and are negative for HER-2/neu gene amplification, limiting use of available targeted therapies. This may translate into poor clinical outcome in this subset of patients as opposed to triple negative subtype of breast cancer. Cancer cell lines have been proved to be a valuable tool in understanding pathogenesis of BCa and development of appropriate drugs. Considering tumor heterogeneity, selecting an appropriate cell line model is imperative to understand signaling mechanisms of each of the distinct sub-types of BCa.Aim of this study was to establish and characterize a BCa cell line from a non-caucasian Pakistani patient. Methodology: Cell line was established from primary culture of tumor tissue from a 65 year old Pakistani female patient diagnosed with T4N1M0 MCa of breast. Specimen was procured in Dulbecco's Modified Eagle's Medium and processed under sterile conditions by mincing and gentle pipetting followed by culturing in complete medium supplemented with 10% fetal bovine serum. Epitheliod colonies were visualized after 3 weeks of culture, which were subsequently passaged and propagated. This cell line has been phenotypically and genotypically characterized using karyotyping, gene expression analysis, immunocytochemistry and florescent insitu hybridization (FISH). Results: Cell line has a human karyotype with multiploidy and population doubling time of 60 hours. Gene expression profiling revealed negative expression of ER, PR and positive expression of androgen receptor (AR) and HER-2/neu. FISH analysis was negative for HER-2/neu amplification. Both basal (cytokeratins 5, 14 &amp; 19) and luminal markers (cytokeratin 8 &amp; 18) are expressed at mRNA level along with myoepithelial markers (CD10, S100A7, p-cadherin, desmin, S100A4, S100A2 &amp; α-SMA). In accordance with mesenchymal phenotype, it has low expression of e-cadherin and high expression of vimentin. Invasion assay revealed this cell line to be non-invasive forming spheroids. CAG repeat length of AR is 22 and AR has been found to be functional by luciferase reporter assay in response to treatment with dihydrotestosterone. Conclusions: We report a novel metaplastic carcinoma cell line from a nan-caucasian female patient with functional AR. Targeting AR signaling may be a promising therapeutic target in this rare sub-type of BCa. Citation Format: Nazia Riaz, Zulfiqar Naqvi, Anwar Ali Siddiqui, Shaista Masood Khan, ElNasir Lalani. Androgen receptor expressing metaplastic carcinoma cell line established from non-caucasian female patient. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1208. doi:10.1158/1538-7445.AM2014-1208

A PILOT STUDY OF HER-2/NEU GENE AMPLIFICATION ASSESSED BY FLUORESCENCE IN SITU HYBRIDIZATION (FISH) IN GASTRIC CANCER

Background: HER-2/neu is a predictive biomarker for treatment of gastric cancer using trastuzumab in combination with chemotherapy. This study aimed to evaluate the status of HER-2/neu gene amplification using fluorescence in situ hybridization (FISH) in gastric cancer. Patients and methods: thirty six gastric cancer patients were assessed HER-2/neu gene amplification by FISH using PathVysionTM HER-2 DNA Probe kit (including HER-2/neu probe and CEP-17 probe) with biopsy and surgical specimens. Results: The HER-2/neu gene amplification was observed in three cases (8.3%), the HER-2/neu gene amplification rate in Lauren’s intestinal-type and diffuse-type were 11.8% and 5.2%, respectively. Conclusion: We applied successfully FISH technique with gastric cancer tissue samples. This technique could be performed as routine test in gastric cancer in order to select patients that benefit from trastuzumab in combination with chemotherapy.

Clinicopathological study on human epidermalgrowth factor receptor-2/neu gene amplification and protein expression in esophageal squamous cell carcinoma

Objective To investigate human epidermalgrowth factor receptor-2 (HER2)/neu gene amplification and protein expression in esophageal squamous cell carcinoma (ESCC) and to explore their clinicopathological correlations.Methods The HER2/neu gene status and protein expression in 120 cases of ESCC were evaluated by using fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC).Results HER2/neu gene amplification and HER2 protein overexpression (+++) were found in 5 (4.17％) and 4 (3.33％) cases of ESCC respectively.HER2 overexpression was associated significantly with HER2/neu gene amplification.There was no correlation between the HER2/neu gene amplification or overexpression of HER2 protein and the ESCC patients' clinicopathological features.Conclusion The HER2/neu gene amplification rate and its protein overexpression rate in ESCC were low,and the role HER2/neu gene plays in ESCC still need further studies. Key words: Esophageal squamous cell carcinoma; Human epidermalgrowth factor receptor-2/neu gene; Immunohistochemistry

High-grade transformation in salivary gland carcinomas

High-grade transformation (HGT) has been described in a variety of salivary gland carcinomas, including acinic cell carcinoma, adenoid cystic carcinoma, epithelial-myoepithelial carcinoma, polymorphous low-grade adenocarcinoma, myoepithelial carcinoma, low-grade mucoepidermoid carcinoma, hyalinizing clear cell carcinoma, and mammary analogue secretory carcinoma, although the phenomenon is a rare event. HGT tumors are composed of conventional carcinomas juxtaposed with areas of HG morphology, usually either poorly differentiated adenocarcinoma or undifferentiated carcinoma, in which the original line of differentiation is no longer evident. The HG component is generally composed of solid nests, sometimes occurring in cribriform pattern of anaplastic cells with large vesicular pleomorphic nuclei, prominent nucleoli and abundant cytoplasm. Frequent mitoses and extensive necrosis are evident. The Ki-67 labeling index is consistently higher in the HG component. p53 abnormalities have been demonstrated in the transformed component in a few examples, but the frequency varies by the histologic type. HER-2/neu over-expression and/or gene amplification is considerably exceptional. The molecular-genetic mechanisms responsible for the pathway of HGT in salivary gland carcinomas largely still remain to be elucidated. Salivary gland carcinomas with HGT have been shown to be more aggressive than conventional carcinomas with a poorer prognosis, suggesting the need for wider resection and neck dissection.