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Chromogenic in Situ Hybridization Technique versus Immunohistochemistry in Assessment of HER2/neu Status in 448 Iraqi Patients with Invasive Breast Carcinoma

BACKGROUND: The rapidly growing knowledge regarding factors controlling tumour growth, with the new modalities of therapy acting on the biological activity of the tumours draw the attention of most cancer researches nowadays and represent a major focus for clinical oncology practice. For the detection of HER2/neu protein overexpression and gene amplification, immunohistochemistry (IHC) and in-situ hybridisation (ISH) is the recommended techniques, respectively, with high concordance between the two techniques. The current United Kingdom recommendations for HER2/neu testing are either for a two-tier system using IHC with reflex ISH testing in equivocal positive cases, or a one-tier ISH strategy. AIM: To compare the results of HER2/neu gene status in patients with breast carcinoma obtained by chromogenic in situ hybridisation with those obtained by immunohistochemistry, and to compare these results with hormonal receptors expression by immunohistochemistry and with age of patients.METHODS: Immunohistochemistry technique was used for evaluation of status of estrogen receptors (ER) and progesterone receptors (PR) and HER2/neu protein expression in 448 Iraqi patients with invasive breast carcinoma with different grades and histological types and then chromogenic in situ hybridization (CISH) technique was applied for all scores of HER2/neu to detect the gene status and compare the results in all negative, equivocal and positive cases by immunohistochemistry (IHC). The cases were referred from different centres, and IHC and CISH techniques were done in central public health laboratory in Baghdad over 28 months, from July 2013 to November 2015. A comparison of the results was made to find the relationship between HER2/neu and hormone receptors status and other clinical parameters like patients age. RESULTS: The mean age of the study cases was 49.08 years, ranging from 24 to 83 years. Of the 448 cases of breast carcinoma, 44 (9.8%) cases were of score 0 by IHC, none of them (0%) showed HER2/neu gene amplification by CISH. 71(15.8%) cases were of score 1 by IHC, 15 (21.12%) of them showed HER2/neu gene amplification by CISH, all were of low amplification. There were 306 (68.3%) cases of score 2 by IHC, of which 102 (33.33%) cases showed HER2/neu gene amplification by CISH, with 79 (25.81%) of them with low amplification and 23 (7.51%) cases with high amplification, while only one case (0.32%) remained in equivocal category. In score 3, all the 27 (6.0%) cases showed gene amplification with 12 (44.44%) cases with low amplification and 15 (55.55) cases with high amplification with overall percentage of gene amplification in score 3 of 100%. There was a significant inverse relationship between hormone receptors (ER and PR) status and HER2/neu gene amplification. No significant relationship was found between the patient’s age and HER2/neu gene amplification.CONCLUSION: Although immunohistochemistry is a widely used, less expensive and reliable test, we strongly advice performance of chromogenic in situ hybridization in assessment of HER2/neu gene status in all cases diagnosed with breast carcinoma as significant number of cases that were reported as negative by immunohistochemistry showed positive amplification by chromogenic in situ hybridization and can get benefit from anti-HER2 targeted treatments.

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Publication Date
Sun Jan 01 2017
Journal Name
Indian Journal Of Pathology And Microbiology
Assessment of topoisomerase II-alpha gene status by dual color chromogenic in situ hybridization in a set of Iraqi patients with invasive breast carcinoma

Abstract Background: The human epidermal growth factor receptor 2(HER2) proto-oncogene is overexpressed or amplified in approximately 15%-25% of invasive breast cancers. Approximately 35% of HER2-amplified breast cancers have coamplification of the topoisomerase II-alpha (TOP2A) gene encoding an enzyme that is a major target of anthracyclines. Hence, the determination of genetic alteration (amplification or deletion) of both genes is considered as an important predictive factor that determines the response of breast cancer patients to treatment. The aims of this study are to determinate TOP2A status gene amplification in a set of Iraqi patients with breast cancer that have had an equivocal (2+) and positive HER2/neu by immunohistochemistry

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Publication Date
Mon Feb 01 2021
Journal Name
International Medical Journal
Use of immunohistochemistry and silver in situ hybridization (Sish) in evaluation of human epidermal growth factor receptor2 (HER2/neu) status in Iraqi patients with breast cancer

Breast cancer is the commonest cause of cancer related death in women worldwide. Amplification or over-expression of the ERBB2 (HER/neu) gene occurs in approximately 15-30% of breast cancer cases and it is strongly associated with an increased disease recurrence and a poor prognosis. Determination of HER2/neu status is crucial in the treatment plan as that positive cases will respond to trastuzumab therapy. It has been used to test for HER2/neu by immunohistochemistry as a first step and then to study only the equivocal positive cases (score 2+) by in situ hybridization technique. The aim of our study is to compare between immunohistochemistry and silver in situ hybridization (SISH) in assessment of human epidermal growth factor (HER2/neu)

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Publication Date
Thu Dec 01 2016
Journal Name
Karbala Journal Of Medicine
Assessment Of Her-2/Neu Gene Amplification Status by Chromogenic in Situ Hybridization in Breast Cancer Patients with Equivocal 2+ Her-2/Neu Immunostaining and Its Relation to The Clinic Pathological Parameters

background: human epidermal growth factor receptor-2 (her2/neu) is related to growth factor receptors with alkaline kinase activity and it is regarded as important prognostic and therapeutic factor that can depended on in breast cancer therapy. HER2/neu expression by immunohistochemistry (IHC) is submitted to a great in terob server inconsistency. Subsequently additional confirmatory tests for assessment of gene alterations and amplification status are needed for patients with early or metastatic breast cancer. In situ hybridization techniques and specifically Chromogenic in situ hybridization (CISH) was arise as a practical, cost-effective, and alternative to fluorescent in situ hybridization in testing for gene alterationAims of the study

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Publication Date
Mon Jan 01 2018
Journal Name
Journal Of Clinical And Diagnostic Research
Immunohistochemical Expression of HER2/neu Receptors in Iraqi Patients with Endometrioid Carcinoma

Introduction: With the advent of era of targeted cancer therapy, the serious side effects of chemoradiotherapy have been minimised. Considering the success story of anti-HER2/neu drugs in breast cancer oncology, the present study was conducted. The study evaluates the immunohistochemical expression of HER2/neu in endometrioid cancer among Iraqi patients. Aim: To assess the immunohistochemical expression of HER2/neu in endometrioid carcinoma of uterus and to find the relationship of this expression with FIGO stage and grade and with patient age. Materials and Methods: In this cross sectional study, formalin-fixed, paraffin-embedded tissue blocks of 62 hysterectomy specimens that were diagnosed as endometrioid carcinoma in the teaching labora

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Publication Date
Thu Jan 14 2016
Journal Name
International Journal For Sciences And Technology
Evaluation of her/2-neu gene status using FISH/CISH techniques in Iraqi breast carcinoma patients

The present study aimed to examine the concordance between FISH/CISH techniques for assessment of amplification of her2neu gene in Iraqi breast carcinoma patients. Seventy four (74) Iraqi breast cancer patients were involved at the study from the Histopathology Department at the Central Public Health Laboratory in Bagdad, Iraq. Amplification of HER2neu was detected in (33.8%) by fluorescence in situ hybridization and (13.51%) showed high amplification by chromogenic in situ hybridization and (32.43%) showed low amplification. The results of chromogenic in situ hybridization were significantly correlated with the results of two-color fluorescence in situ hybridization with the same tumors. In addition, the study involved the correlation betw

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Publication Date
Sun Apr 01 2012
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Detection of Human papillomavirus in surface epithelial ovarian carcinoma using in situ hybridization technique

Background: The role of Human papillomaviruses (HPV) in the etiology of ovarian cancer remains unclear and the results are controversial. Several studies have verified the presence of HPV DNA in both malignant and benign ovarian tumors.
Objectives: Determine the percentage of detection of HPV high (16&18) and low risk types (6&11) in surface epithelial ovarian carcinoma compared to benign and control groups.
Materials And Methods: Molecular detection and genotyping of HPV DNA were performed in 76 ovarian tissue blocks by using in situ hybridization (ISH) technique for detecting and localization of high risk HPV (16 and 18) and low risk HPV (6&11) types.
Results: The presence of ISH signals fo

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Publication Date
Thu Sep 30 2021
Journal Name
Iraqi Journal Of Science
Chromogenic in Situ Hybridization for Human Cytomegalovirus-DNA Detection in Tissue Subsets with Prostatic Adenocarcinoma and Benign Hyperplasia

     Human cytomegalovirus (HCMV) infects a wide range of human cells, resulting in both benign and malignant tumors. In the last few decades, proteins and/or nucleic acids of the virus were found to be often highly expressed in in patients with basal cell hyperplasia and prostatic neoplasia.

This research aimed to unravel the rate of HCMV infections among prostatic tissue subsets from Iraqi patients with adenocarcinoma and benign hyperplasia.

One hundred, formalin-fixed and paraffin embedded prostatic tissues were obtained from 40 tissue samples collected from different grades of prostate carcinoma; 40 from benign prostatic hyperplasia and 20 from apparently healthy prostatic tissues. These tissue spe

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Publication Date
Wed Apr 01 2015
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Detection of Human Papilloma Virus type 6 and type 11 in women with Breast Cancer by in situ hybridization technique.

Background: Breast cancer is one of the common malignancies among women worldwide. Human papillomavirus (HPV) infections have been linked to many human cancers in addition to cervical cancer and one of them is breast cancer.
Objective: To investigate the presence of human papilloma virus type 6 and type 11in breast cancer tissue specimens by in situ hybridization technique.
Patients and Methods: Thirty four formalin-fixed, paraffin embedded tissue blocks from breast cancer patients were obtained from the archives of the pathology laboratory of Al-Yarmouk Teaching Hospital from January 2011 to July 2012. In addition formalin-fixed, paraffin embedded blocks tissue for twenty fibroadenoma of breast were collected and used as control g

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Publication Date
Tue Jan 02 2018
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Breast Cancer Subtypes among Iraqi Patients: Identified By Their ER, PR and HER2 Status

Background: Breast cancer ranks the first among the Iraqi population since three decades and is currently forming a major public health problem being the second cause of death women. Novel management of breast cancer depends upon precise evaluation of their molecular subtypes; identified by Hormone (Estrogen and Progesterone) receptors and HER2 contents of the primary tumor.
Objective: To assess the rates of the different molecular breast cancer subtypes in the examined tissue specimens belonging to females diagnosed with breast cancer in Iraq; correlating the findings with those reported in the literature at the regional and global levels.
Patients and Methods: This retrospective study documented the findings of tissue biopsy exam

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Publication Date
Mon Apr 01 2019
Journal Name
Biochemical And Cellular Archives
Association Of Human Cytomegalovirus With Her2 Protooncogene Overexpression In Iraqi Breast Cancer Patients

Human cytomegalovirus (HCMV) has a worldwide distribution and common infections. The presence of HCMV genome and antigens has been detected in many kinds of human cancers. The proto-oncogene Her2/neu is overexpressed in 25% to 30% of human breast cancers and frequently associated with tumor aggressiveness and worse prognosis. Examination was applied to detect different HCMV antigens and to demonstrate their correlation with Her2/neu overexpression in breast cancer. The present study includes samples from 70 women of 60 breast cancer patients and 10 normal breast tissues. Formalin-fixed paraffin embedded tissue blocks were obtained from each woman according to ethical approval. Human cytomegalovirus early antigen expression was detected in 5

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