Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Rosa, Francisco Dalla; Santos, Renata Ferreira dos; Rogatto, Sílvia Regina; Domingues, Maria Aparecida Custódio

doi:10.1590/1414-431x20132483

Cited by 23 publications

(23 citation statements)

References 73 publications

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“…Owing to its resemblance to immunohistochemistry staining (57), CISH is also easier to interpret by pathologists who are not trained in fluorescence microscopy. Furthermore, in previous studies, CISH was observed to be well-correlated with FISH (46,(58)(59)(60). The present study used standard CISH to demonstrate that HER2 was amplified in Iranian (Tehran province) patients with PCa.…”

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confidence: 66%

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HER2 gene amplification in patients with prostate cancer: Evaluating a CISH-based method

et al. 2016

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Abstract. Prostate cancer (PCa) is one of the most widespread malignancies in the world. The role of the human epidermal growth factor receptor 2 (HER2) in the pathogenesis and progression of human PCa remains poorly understood. In contradiction with breast cancer, studies on HER2 overexpression and gene amplification in PCa have produced varying results, although the HER2 oncogene has been implicated in the biology of numerous tumor types, and serves as a prognostic marker and therapeutic target in breast cancer. Technical challenges are considered the main reasons for data discrepancies. Amplification of the HER2 gene has previously been reported in PCa, in which it was associated with tumor progression. The present study aimed to evaluate the prevalence and clinical significance of HER2 amplification in PCa. A total of 32 biopsy samples obtained from human prostate adenocarcinomas were evaluated by chromogenic in situ hybridization (CISH) to determine the frequency of patients with HER2 gene amplifications. High copy numbers of HER2 were detected in 19 of the prostate tumors analyzed. The results of the present study suggested that, in patients without amplification of HER2, high levels of prostate-specific antigen or a high Gleason score were not significantly correlated with a high pathologic stage. Furthermore, amplification levels of the HER2 gene were directly associated with pathologic stage in patients with PCa. Therefore, the potential use of HER2 as a prognostic factor or therapeutic target for PCa warrants further study. IntroductionProstate cancer (PCa), a common non-skin, sex-limited cancer, is the second cause of cancer-associated mortality (after lung cancer) in the USA (1,2) and the second most prevalent cancer among Iranian men (3). Worldwide, PCa is the second most commonly diagnosed cancer and, according to the International Agency for Research on Cancer's GLOBOCAN 2012 (4) database, it is the fifth leading cause of cancer-associated mortality in men. The incidence of PCa is increasing worldwide, although there is a marked variation in its incidence among different regions (5). The clinical configuration of PCa has noticeably altered over the past few years. As a localized disease, it is easily treated by radical radiation therapy or a prostatectomy; however, if the tumor becomes malignant, it transforms into a life-threatening disease (6).The progression and application of novel high-resolution technologies has enhanced the detection of genomic alterations, enabling elucidation of the complex nature and heterogeneity of PCa (7). The differentiation of PCa tumors is typically based on the serum expression levels of prostate-specific antigen (PSA), although, in certain cases, PSA levels do not accurately reflect tumor burden (8). Previous studies have identified a number of genetic, epigenetic and environmental risk factors for PCa (9-11). Among them, genetic aberrations and chromosomal changes have been suggested to serve a significant role in the development and progression of PCa (12)

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confidence: 66%

“…Permanent staining and the absence of a fluorescent dye make CISH a suitable replacement for FISH (69). In addition, its usability, relative inexpensiveness and speed make CISH more attractive than FISH for assessing HER2 amplification/overexpression (59,70).…”

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confidence: 99%

HER2 gene amplification in patients with prostate cancer: Evaluating a CISH-based method

et al. 2016

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“…First, IHC scores vary depending on the antibodies used [9]. Second, IHC is sensitive to pre-analytical variation, including tissue fixation [31].…”

Section: Discussionmentioning

confidence: 99%

“…These two techniques, demonstrating excellent concordance [9], are less sensitive to pre-analytical variation than IHC, but they are labor-intensive, expensive and somewhat difficult to interpret.…”

Section: Introductionmentioning

confidence: 99%

Added Value of HER-2 Amplification Testing by Multiplex Ligation-Dependent Probe Amplification in Invasive Breast Cancer

Kuijpers¹,

Moelans

Slooten³

et al. 2013

PLoS ONE

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BackgroundHER-2 is a prognostic and predictive marker, but as yet no technique is perfectly able to identify patients likely to benefit from HER-2 targeted therapies. We aimed to prospectively assess the added value of first-line co-testing by IHC, and multiplex ligation-dependent probe amplification (MLPA) and chromogenic in situ hybridization (CISH).MethodsAs local validation, HER-2 MLPA and CISH were compared in 99 breast cancers. Next, we reviewed 937 invasive breast cancers, from 4 Dutch pathology laboratories, that were prospectively assessed for HER-2 by IHC and MLPA (and CISH in selected cases).ResultsThe validation study demonstrated 100% concordance between CISH and MLPA, if both methods were assessable and conclusive (81.8% of cases). Significant variation regarding percentages IHC 0/1+ and 2+ cases was observed between the laboratories (p<0.0001). Overall concordance between IHC and MLPA/CISH was 98.1% (575/586) (Kappa = 0.94). Of the IHC 3+ cases, 6.7% failed to reveal gene amplification, whereas 0.8% of the IHC 0/1+ cases demonstrated gene amplification. Results remained discordant after retrospective review in 3/11 discordant cases. In the remaining 8 cases the original IHC score was incorrect or adapted after repeated IHC staining.ConclusionsMLPA is a low-cost and quantitative high-throughput technique with near perfect concordance with CISH. The use of MLPA in routinely co-testing all breast cancers may reduce HER-2 testing variation between laboratories, may serve as quality control for IHC, will reveal IHC 0/1+ patients with gene amplification, likely responsive to trastuzumab, and identify IHC 3+ cases without gene amplification that may respond less well.

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“…These methods are used worldwide to test for HER2 overexpression/amplification within routine pathology. However, HER2 IHC analysis is influenced by several factors including fixation time, antigen retrieval and antibody specificity and sensitivity (6,(19)(20)(21)(22). Furthermore, the interpretation of HER2 IHC is semi-quantitative and subjective, which may lead to inter-observer variability affecting the accuracy of the results (23)(24)(25).…”

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Evaluation of analytical accuracy of HER2 status in patients with breast cancer

Jensen

Thomas

Christensen

et al. 2020

APMIS

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Human epidermal growth factor receptor 2 (HER2) gene status and overexpression, occurring in~13.6% of primary breast cancers, is essential for identifying patients likely to benefit from biological treatment. In this method of evaluation study, we tested and compared the HER2 gene-protein assay (GPA) with routine HER2 immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). The GPA was evaluated using 67 formalin-fixed paraffin-embedded (FFPE) HER2 equivoval IHC (2+) breast cancer tissue samples. Overall, agreement between GPA silver in situ hybridization (SISH) and FISH was 91.9% (57/62). Regression analysis revealed slightly higher, but non-significant difference in HER2/chromosome enumeration probe 17 (CEP17) ratio for GPA as compared to FISH (p = 0.074). Intraclass correlation coefficients (ICCs) of 0.94 and Spearman´s rank correlation coefficients of 0.93 (p < 0.0001) for FISH and GPA SISH suggested strong inter-observer association for methods with one observer counting on average 0.23 significant higher for GPA SISH (p = 0.014). Intra-observer IHC method reproducibility was 52.6% (j = 0.3122, p = 0.004) and 79.7% (j = 0.6428, p = 0.9197), suggesting fair significant and substantial non-significant difference between tests for reviewers. Inter-observer reproducibility for IHC methods was 53%. While inter-observer reproducibility for experienced IHC interpretation suggested significant differences (j = 0.3636, p = 0.0332), unexperienced interpretation of IHC GPA suggested fair non-significant difference between reviewers (j = 0.3101, p = 0.0747). Using FISH as reference, the diagnostic indices for GPA SISH were as follows: sensitivity 100%, specificity 95% and accuracy 92%. Inaccuracy between tests was in 80% of cases due to ISH categorization as equivocal by one of the methods. IHC results highlight that it may be beneficial with a method for simultaneously visualization of HER2 gene and protein status.

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Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Cited by 23 publications

References 73 publications

HER2 gene amplification in patients with prostate cancer: Evaluating a CISH-based method

HER2 gene amplification in patients with prostate cancer: Evaluating a CISH-based method

Added Value of HER-2 Amplification Testing by Multiplex Ligation-Dependent Probe Amplification in Invasive Breast Cancer

Evaluation of analytical accuracy of HER2 status in patients with breast cancer

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