Laboratory assessment of the status of Her-2/neu protein and oncogene in breast cancer specimens: comparison of immunohistochemistry assay with fluorescence in situ hybridisation assays

Wang, S; Saboorian, M. Hossein; Frenkel, Eugene P.; Hynan, Linda S.; Gokaslan, Sefik T.; Ashfaq, Raheela

doi:10.1136/jcp.53.5.374

Cited by 162 publications

(114 citation statements)

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“…The specimens were fixed in 10% neutral buffered formalin and then paraffin embedded as described elsewhere (9). After histologic diagnosis was made, one representative tumor tissue block from each of the infiltrating breast carcinoma specimens was evaluated for the HER-2/neu status.…”

Section: Specimen Procurement and Processingmentioning

confidence: 99%

“…The IHC assay for HER-2/neu protein has been described elsewhere (9). Briefly, the paraffinembedded tissue blocks were cut to 3-m sections that were then deparaffinized and heat treated for antigen retrieval.…”

Section: Ihc Assaymentioning

confidence: 99%

“…Upon completion of the IHC staining, all slides were first quantitated using the manual method as described elsewhere (9). The cytoplasmic membrane staining of the invasive cancer component was graded as negative, low (1ϩ), medium (2ϩ), or high positive (3ϩ; 9).…”

Section: Ihc Assaymentioning

confidence: 99%

“…A FISH assay, PathVysion, was used to evaluate the genetic variables, including HER-2/neu gene and chromosome 17 copy number, as previously described (9). Briefly, the PathVysion assay was performed using the manufacturer's recommended protocol on 4-m sections prepared from paraffinembedded tissue blocks.…”

Section: Fish Assaymentioning

confidence: 99%

“…The other 151 specimens (accessioned between March 1999 and March 2000) contained all cases with positive IHC staining (1ϩ to 3ϩ, total 136 cases) and only 15 random cases with negative IHC staining. The exclusion of most cases with negative IHC staining was based on the previous finding that none of such cases had HER-2/neu gene amplification (9). This selection process resulted in underrepresentation of the IHC-negative specimens for the FISH assay.…”

Section: Case Selectionmentioning

confidence: 99%

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Aneusomy 17 in Breast Cancer: Its Role in HER-2/neu Protein Expression and Implication for Clinical Assessment of HER-2/neu Status

et al. 2002

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HER-2/neu protein overexpression in breast cancer is mostly caused by HER-2/neu gene amplification. However, it is unclear whether aneusomy 17 may also play a role. Using immunohistochemistry assay (IHC) with DAKO antibody and manual quantitation, 189 specimens were selected from archival invasive breast cancer specimens, including most IHC-positive and some IHC-negative cases (n ‫؍‬ 158 and 31, respectively).

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Section: Specimen Procurement and Processingmentioning

confidence: 99%

Section: Ihc Assaymentioning

confidence: 99%

Section: Ihc Assaymentioning

confidence: 99%

Section: Fish Assaymentioning

confidence: 99%

Section: Case Selectionmentioning

confidence: 99%

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Aneusomy 17 in Breast Cancer: Its Role in HER-2/neu Protein Expression and Implication for Clinical Assessment of HER-2/neu Status

et al. 2002

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Amplification of the HER-2/neu oncogene is uncommon in pediatric osteosarcomas

Maitra

Wanzer

Weinberg

et al. 2001

Cancer

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BACKGROUND Recent reports have described overexpression of p185c‐erbB2, the product of the HER‐2/neu oncogene, in more than 40% of archival osteosarcomas that were examined by immunohistochemistry (IHC). However, IHC can be influenced by the method of fixation, extent of antigen retrieval, specificity and sensitivity of the antibody clone, and the use of an arbitrary semiquantitative scoring system. In contrast, fluorescence in situ hybridization (FISH) assays that assess HER‐2/neu gene copy numbers in individual cells are more consistent, more reproducible, and less subjective than IHC. METHODS The authors examined pretreatment nondecalcified archival tissue from 21 high‐grade pediatric osteosarcomas for amplification of the HER‐2/neu oncogene using an Food and Drug Administration (FDA)‐approved FISH assay (PathVysion, Vysis Inc., Downers Grove, IL). Additionally, IHC for p185c‐erbB2 was performed in all cases using the Dako polyclonal antibody clone A0485 (Dako Co., Carpinteria, CA). RESULTS None of the 21 osteosarcomas had evidence of HER‐2/neu gene amplification by FISH, whereas p185c‐erbB2 IHC was negative in all cases. CONCLUSIONS HER‐2/neu gene amplification appeared to be an uncommon event in pediatric osteosarcomas. The reason(s) for discordance between previous IHC data and the current FISH and IHC results was unknown, but might reflect intrinsic variations in antibody clones, or might suggest that, in some cases, the occurrence of protein overexpression is independent of gene amplification. Cancer 2001;92:677–83. © 2001 American Cancer Society.

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c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein over-expression in operable breast-cancer patients

Riou¹,

Mathieu²,

Barrois³

et al. 2001

Int. J. Cancer

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We investigated the effect of hepatocyte-derived extracellular matrix (ECM) on the expression of erb-B2 and erb-B3 in colon cancer cell lines, as well as the role of erb-B2 and erb-B3 in colon cancer cell proliferation. Colon cancer cell lines plated on hepatocyte-derived ECM had increased protein levels of both erb-B2 and erb-B3. The addition of soluble recombinant proteoglycan syndecan-4 also resulted in higher expression of erb-B2 and erb-B3. We prepared hepatocytederived ECM from 1 to 7 days' cultures of hepatocytes, which contained different amounts of sulfated glycosaminoglycans. There was a direct correlation between the amounts of sulfated glycosaminoglycans in the ECM and the levels of erb-B2 and erb-B3 in the colon cell line KM12. The stimulatory effect of hepatocyte-derived ECM was abolished when the colon cancer cells were cultured in the presence of antibodies to erb-B2. These studies show that hepatocyte-derived ECM and the heparan sulfate proteoglycans present in it are responsible for inducing erb-B2 and erb-B3 in colon cancer cells. The growth stimulatory effect of extracellular matrix is mediated, at least in part, by increased expression of erb-B2 and erb-B3. © 2001 Wiley-Liss, Inc. Key words: extracellular matrix; colon cancer; heparan sulfateThe local milieu of various organs colonized by metastatic tumor cells, in particular the extracellular matrix (ECM) of those organs, plays a central role in site-specific metastasis. The phenomenon of site-specific metastasis was described as early as 1889, when Paget suggested the "seed and soil" hypothesis, proposing that the newly colonized organ must be a fertile soil for tumor cells. The metastatic cells need to recognize and attach to the local ECM components and to the cells of the organ. 1,2 The local ECM can also affect tumor cell proliferation by various mechanisms, one of which is the induction of autocrine growth factors and growth factor receptors.Previously we have shown that the ability of hepatomas and breast carcinomas to metastasize to a specific organ in vivo correlated directly with the ability of the cells to survive at clonal density in vitro on ECM derived from that organ. 3 We have found that the ECM component that affected the clonal growth efficiency in hepatomas and breast cancer cell lines was heparin. 3 Our studies suggested that organ-specific proteoglycans and their glycosaminoglycan chains were responsible for the regulation of autocrine growth factors in metastatic cells. 4 The autocrine growth factors could trigger either proliferation or differentiation signals, 5 depending on the differentiated state of the tumor cells and on the ECM components present.In colorectal tumors, one of the key growth factor families regulating cell proliferation is the epidermal growth factor (EGF) family, comprising both growth factors and growth factor receptors. The number of EGF receptors on colon cancer cells correlated with increased metastasis to the liver. 6 Erb-B2, another member of EGF receptor family, was found to be amplified i...

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Laboratory assessment of the status of Her-2/neu protein and oncogene in breast cancer specimens: comparison of immunohistochemistry assay with fluorescence in situ hybridisation assays

Cited by 162 publications

References 17 publications

Aneusomy 17 in Breast Cancer: Its Role in HER-2/neu Protein Expression and Implication for Clinical Assessment of HER-2/neu Status

Aneusomy 17 in Breast Cancer: Its Role in HER-2/neu Protein Expression and Implication for Clinical Assessment of HER-2/neu Status

Amplification of the HER-2/neu oncogene is uncommon in pediatric osteosarcomas

c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein over-expression in operable breast-cancer patients

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