Reverse transcription-polymerase chain reaction assay for multiple mRNA markers in the detection of breast cancer metastases in sentinel lymph nodes

Manzotti, Michela; Dell’Orto, Patrizia; Maisonneuve, Patrick; Zurrida, Stefano; Mazzarol, Giovanni; Viale, Giuseppe

doi:10.1002/1097-0215(20010920)95:5<307::aid-ijc0153>3.0.co;2-q

Cited by 69 publications

(46 citation statements)

References 34 publications

(94 reference statements)

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“…Therefore, a tumour-free sentinel lymph node implies the absence of lymph node metastases in the entire lymphatic basin. This sentinel node concept had been previously confirmed by H&E examinations, immunohistochemical study and nonquantitative RT -PCR assays (Giuliano et al, 1994;Albertini et al, 1996;Veronesi et al, 1997;Krag et al, 1998;Kataoka et al, 2000;Manzotti et al, 2001;Branagan et al, 2002;Ishida et al, 2002).…”

mentioning

confidence: 61%

“…Recently, RT -PCR assays with tumour-specific gene markers such as CK19, CEA, MUC1, PIP, Mapsin and mammaglobin B have been widely used to detect occult metastasis in cancer patients (Noguchi et al, 1994(Noguchi et al, , 1996Schoenheld et al, 1994;Mori et al, 1995Mori et al, , 1998Kataoka et al, 2000;Masuda et al, 2000;Manzotti et al, 2001;Mitas et al, 2001). To date, however, no marker that is expressed in all breast cancer tissues, but not in normal lymph node has been available, partly because of the heterogeneity of marker gene expression in cancer cells.…”

Section: Discussionmentioning

confidence: 99%

“…To date, however, no marker that is expressed in all breast cancer tissues, but not in normal lymph node has been available, partly because of the heterogeneity of marker gene expression in cancer cells. Therefore, multimarker analysis will probably be necessary for the precise and accurate examination of lymph node metastasis (Manzotti et al, 2001;Mitas et al, 2001). On the other hand, some evidence suggests that false positives may arise due to several factors, including illegitimate CK19 mRNA expression in lymphocytes, the presence of noncancerous epithelial cells in lymph nodes and the use of inappropriate primer modification and too many PCR amplification cycles (Bostic et al, 1998b).…”

Section: Discussionmentioning

confidence: 99%

“…Several reports have indicated that the detection of micrometastases using reverse transcriptase -polymerase chain reaction (RT -PCR) assay may be clinically important (Noguchi et al, 1994(Noguchi et al, , 1996Schoenheld et al, 1994;Mori et al, 1995Mori et al, , 1998Masuda et al, 2000;Manzotti et al, 2001). Moreover, screening for metastatic disease in axillary lymph nodes by RT -PCR amplification has been shown to be more sensitive and cost effective than serial sectioning and immunohistochemical staining (Lockett et al, 1998).…”

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confidence: 99%

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Quantitative evaluation of metastases in axillary lymph nodes of breast cancer

et al. 2003

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We have established a highly sensitive and quantitative reverse transcriptase -polymerase chain reaction (RT -PCR) method to detect axillary lymph node metastases of breast cancer. Amplifying cytokeratin 19 (CK19) mRNA transcripts using real-time TaqMan PCR made it possible to quantify axillary metastatic burden. Metastases in 358 axillary lymph nodes obtained from 23 breast cancers of 22 patients were investigated by conventional haematoxylin and eosin (H&E) staining, immunohistochemical staining and quantitative RT -PCR assay. The detection rates of axillary lymph node metastasis using H&E staining, immunohistochemistry and RT -PCR assay were 4.5, 5.9 and 13.1%, respectively. RT -PCR assay was the most sensitive of these three methods for detecting lymph node metastases. Cytokeratin 19 mRNA expression values of both histologically and immunohistochemically positive lymph nodes were significantly higher than the values for lymph nodes judged to be negative by both histological and immunohistochemical methods (Po0.0001), and those of histologically negative, but immunohistochemically positive lymph nodes were significantly higher than the values for lymph nodes judged to be negative by both histological and immunohistochemical methods (Po0.0001). Furthermore, metastatic rates of sentinel nodes were higher than the rates of nonsentinel lymph nodes as measured by all three methods. These results indicate that quantitative RT -PCR assay is a sensitive and reliable method for detecting lymph node metastasis. Furthermore, quantification of metastases in sentinel lymph nodes by quantitative RT -PCR assay may be useful to assess the entire axillary burden of breast cancer patients.

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confidence: 61%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Quantitative evaluation of metastases in axillary lymph nodes of breast cancer

et al. 2003

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“…The two specific markers identified, mammaglobin and CK19, have been previously published as potentially useful components of multimarker panels for the identification of occult metastasis in breast lymph nodes. 12,32,33 However, previous studies have also concluded that both markers are expressed in the lymph nodes of 20 to 40% of patients categorized as lymph node negative by standard histological analysis. 10,11 It has also been reported that lymph nodes from a high percentage of cancer-free patients express CK19.…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of Optimal Gene Expression Markers for Detection of Breast Cancer Metastasis

Backus¹,

Laughlin²,

Wang³

et al. 2005

The Journal of Molecular Diagnostics

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Sentinel lymph node (SLN) status is highly predictive of overall axillary lymph node involvement in breast cancer. Historically, SLN-positive patients have undergone axillary lymph node dissection in a second surgery. Intraoperative SLN analysis could reduce the cost and complications of a second surgery; however, existing histopathological methods lack standardization and exhibit poor sensitivity. Rapid molecular methods may lead to improved intraoperative diagnosis of SLN metastasis. In this study, we used a genome-wide gene expression analysis of breast and other tissues to identify seven putative markers for detecting breast cancer metastasis. We assessed the utility of these markers for identifying clinically actionable metastases in lymph nodes through reverse transcriptase-polymerase chain reaction analysis of SLNs from 254 breast cancer patients. Polymerase chain reaction signals were compared to pathology on a per-patient basis. The optimal two-gene combination, mammaglobin and cytokeratin 19, detected clinically actionable metastasis in breast SLNs with 90% sensitivity and 94% specificity. Application of stringent criteria for identifying presumptive hematoxylin-and eosin-positive samples increased sensitivity and specificity to 91 and 97%, respectively. This study represents the first comprehensive demonstration of the utility of gene expression markers for detecting clinically actionable breast metastases. An intraoperative molecular assay using these markers has the potential to significantly reduce second surgeries for patients undergoing SLN dissection. Breast cancer is second only to lung cancer in mortality among women worldwide.1 In the care of this significant disease, the evaluation of blood, bone marrow, and lymph nodes for the presence of metastatic cells is an important component of disease characterization and management.2-6 The method for assessment of these peripheral tissues is largely dependent on histological and cytological methods. Detection of metastasis in lymph nodes is typically accomplished by hematoxylin and eosin (H&E) and antibody staining of lymph node sections.7,8 Analysis of bone marrow samples, although still in development, currently involves antibody staining of cytological smears. For some time, there has been discussion about the potential for the use of molecular biological tools to supplement or to improve existing methods.9 Molecular methods such as reverse transcriptase-polymerase chain reaction (RT-PCR) offer increased analytical sensitivity compared with standard histological methods. 10 -12 In addition, nucleic acid-based detection methods such as real-time PCR offer the potential of rapid and sensitive point-of-care testing and the application of objective quantitative assay cut-offs.A clear intersection between these features is the intraoperative assessment of sentinel lymph nodes (SLNs) for the presence of clinically actionable metastasis, a level of metastasis that would be expected to consistently lead to a subsequent axillary lymph node (ALN) dissecti...

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Intraoperative assessment of sentinel lymph nodes in breast cancer

Layfield¹,

Agrawal

Roché

et al. 2011

Journal of British Surgery

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Background: Sentinel lymph node biopsy (SLNB) reduces the morbidity of axillary clearance and is the standard of care for patients with clinically node-negative breast cancer. The ability to analyse the sentinel node during surgery enables a decision to be made whether to proceed to full axillary clearance during primary surgery, thus avoiding a second procedure in node-positive patients.Methods: Current evidence for intraoperative sentinel node analysis following SLNB in breast cancer was reviewed and evaluated, based on articles obtained from a MEDLINE search using the terms 'sentinel node', 'intra-operative' and 'breast cancer'.Results and conclusion: Current methods for evaluating the sentinel node during surgery include cytological and histological techniques. Newer quantitative molecular assays have been the subject of much recent clinical research. Pathological techniques of intraoperative SLNB analysis such as touch imprint cytology and frozen section have a high specificity, but a lower and more variably reported sensitivity. Molecular techniques are potentially able to sample a greater proportion of the sentinel node, and could have higher sensitivity.

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Reverse transcription-polymerase chain reaction assay for multiple mRNA markers in the detection of breast cancer metastases in sentinel lymph nodes

Cited by 69 publications

References 34 publications

Quantitative evaluation of metastases in axillary lymph nodes of breast cancer

Quantitative evaluation of metastases in axillary lymph nodes of breast cancer

Identification and Characterization of Optimal Gene Expression Markers for Detection of Breast Cancer Metastasis

Intraoperative assessment of sentinel lymph nodes in breast cancer

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