Chromogenic and fluorescent in situ hybridization in breast cancer

Lambros, Maryou B.; Natrajan, Rachael; Reis‐Filho, Jorge S.

doi:10.1016/j.humpath.2007.04.011

Cited by 85 publications

(76 citation statements)

References 170 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Instead of the fluorogens used in FISH, this technique uses chromogens. This has several advantages 24 : where FISH requires a fluorescence microscope for interpretation, CISH can be interpreted using a normal bright field microscope. CISH allows analysis of tumour morphology, making it possible to interpret tumour heterogeneity and gene copy number in different components of the tumour (an invasive and an in situ component).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer

et al. 2009

View full text Add to dashboard Cite

HER2 overexpression in breast cancer is associated with worse clinical outcome. To select patients for antiHer2-based therapy immunohistochemistry is commonly performed as a first step to assess Her2 status. However, interobserver and interlaboratory variability can significantly compromise adequate assessment of Her2 status. In addition, immunohistochemistry does not always result in an unambiguous test result requiring additional testing for Her2 gene amplification. This study aimed to improve the reliability of Her2 immunohistochemistry by using rabbit monoclonal antibody 4B5 as an alternative to mouse monoclonal antibody CB11 routinely used in our laboratory. Therefore, 283 breast adenocarcinomas were included in a tissue microarray. Immunohistochemistry using the 4B5 and CB11 antibodies, and fluorescence and chromogenic in situ hybridization (FISH or CISH) were performed. Immunohistochemistry was scored by two independent investigators. We found that 4B5 staining was more distinct than CB11 staining. For CB11 staining, there were 12% (BV) and 5% (JW) 2 þ scores compared with 4% (BV) and 2% (JW) for 4B5. There was a strong trend towards higher interobserver agreement for 4B5 compared with CB11 (4B5: j 0.87, 95% CI 0.79-0.96; CB11: j 0.77, 95% CI 0.66-0.88). There were no significant differences in sensitivity, specificity and predictive values between CB11 and 4B5. Our results indicate that the 4B5 antibody provides more robust assessment of immunohistochemical Her2/neu status and will reduce the number of gene amplification tests compared with CB11. However, for tumours with a 2 þ score additional gene amplification measurement using FISH or CISH remains necessary.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Many authors have compared CISH with FISH (reviewed by Lambros et al 24 ). Most studies find an agreement of both methods of more than 90%.…”

Section: Discussionmentioning

confidence: 99%

Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer

et al. 2009

View full text Add to dashboard Cite

show abstract

“…18 Amplification of MYC has been associated with poor prognosis and resistance to anti-estrogen therapy. 19 Therapeutic or prognostic significance of other frequently amplified genes such as cyclin D1 (CCND1) 20 or frequent loss of genes such as E-cadherin (CDH1) is less clear, and comparative genomic hybridization (CGH) studies have pointed to many more genes and chromosomal loci with potentially important copy number changes. [21][22][23] Nevertheless, no single gene copy number seems to completely explain prognosis or response to therapy of individual breast cancer patients.…”

mentioning

confidence: 99%

Molecular profiling of invasive breast cancer by multiplex ligation-dependent probe amplification-based copy number analysis of tumor suppressor and oncogenes

et al. 2010

View full text Add to dashboard Cite

Several oncogenes and tumor-suppressor genes have been shown to be implicated in the development, progression and response to therapy of invasive breast cancer. The phenotypic uniqueness (and thus the heterogeneity of clinical behavior) among patients' tumors may be traceable to the underlying variation in gene copy number of these genes. To obtain a more complete view of gene copy number changes and their relation to phenotype, we analyzed 20 breast cancer-related genes in 104 invasive breast cancers with the use of multiplex ligation-dependent probe amplification (MLPA). We identified MYC gene amplification in 48% of patients, PRDM14 in 34%, topoisomerase IIa (TOP2A) in 32%, ADAM9 in 32%, HER2 in 28%, cyclin D1 (CCND1) in 26%, EMSY in 25%, IKBKB in 21%, AURKA in 17%, FGFR1 in 17%, estrogen receptor alpha (ESR1) in 16%, CCNE1 in 12% and EGFR in 9% of patients. There was a significant correlation between the number of amplified genes and the histological grade and mitotic index of the tumor. Gene amplifications of EGFR, CCNE1 and HER2 were negatively associated with estrogen receptor status whereas FGFR1, ADAM9, IKBKB and TOP2A revealed a positive association. Amplifications of ESR1, PRDM14, MYC and HER2 were associated with a high mitotic index, and PRDM14 and HER2 amplifications with high histological grade. MYC amplification was detected more frequently in ductal tumors and high-level MYC amplifications were significantly associated with large tumor size. HER2/MYC, HER2/CCNE1 and EGFR/MYC co-amplified tumors were significantly larger than tumors with either of these amplifications. Gene loss occurred most frequently in E-cadherin (CDH1) (20%) and FGFR1 (10%). In conclusion, MLPA analysis with this 'breast cancer kit' allowed to simultaneously assess copy numbers of 20 important breast cancer genes, providing an overview of the most frequent (co)amplifications as well as interesting phenotypic correlations, and thereby data on the potential importance of these genes in breast cancer.

show abstract

“…At least 30, non-equivocal and nonoverlapping neoplastic cells were counted per case. Nonamplified cases were defined as those with one to five signals per nucleus in >50% of tumor cells; amplification was defined as i) more than 5 gene copies per nucleus in >50% of tumor cells, ii) when small or large gene copy clusters were found in >50% of tumor cells 12 .…”

Section: Cish Analysismentioning

confidence: 99%

Rabbit antibodies for hormone receptors and HER2 evaluation in breast cancer

Nunes¹,

Sanches²,

Rocha³

et al. 2009

Rev. Assoc. Med. Bras.

View full text Add to dashboard Cite

SUMMARYBACKGROUND. Novel rabbit monoclonal antibodies (RabMab) for estrogen (ER), progesterone (PR) receptors and HER2 evaluation by immunohistochemistry have recently been commercially released. We compared the RabMab anti-ER, anti-PR and anti-HER2 to mouse monoclonal antibodies (Mab) using tissue microarrays (TMA) of breast carcinomas. METHODS. Two TMA containing breast carcinomas were built. Sections were immunostained using anti-ER and anti-PR, Mab and RabMab. The sections stained for ER and PR were evaluated considering positive those tumors in which more than 1% of the tumor cell nuclei stained moderate or strong. For HER2, the immunostained sections were evaluated using the ASCO/CAP guidelines for HER2. Chromogenic in situ hybridization (CISH) was used as the gold standard for HER2 evaluation. CISH was evaluated using the Zymed HER2 CISH interpretation guidelines. RESULTS. RabMab against ER have similar staining patterns compared to the 6F11 (Mab), but stronger than 1D5 (Mab) from three different suppliers. The RabMab against PR provide stronger and sharper immunohistochemical signals compared to Mab. The detection of HER2 protein overexpression was more prevalent with the polyclonal antibodies and RabMab than with the Mab. These were more specific than the RabMab, which were more sensitive when compared to CISH. CONCLUSION. The novel RabMab against ER and PR showed higher intensity of staining than the Mab. The RabMab against HER2 is more sensitive than Mab, however, Mab presented more specificity than RabMab when compared to CISH for HER2 evaluation of breast carcinomas.

show abstract

Chromogenic and fluorescent in situ hybridization in breast cancer

Cited by 85 publications

References 170 publications

Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer

Validation of the 4B5 rabbit monoclonal antibody in determining Her2/neu status in breast cancer

Molecular profiling of invasive breast cancer by multiplex ligation-dependent probe amplification-based copy number analysis of tumor suppressor and oncogenes

Rabbit antibodies for hormone receptors and HER2 evaluation in breast cancer

Contact Info

Product

Resources

About