Detection of micrometastatic breast cancer by means of real time quantitative RT‐PCR and immunostaining in perioperative blood samples and sentinel nodes

Schröder, Carolien P.; Ruiters, Marcel H. J.; Jong, Steven de; Tiebosch, A.T.M.G.; Wesseling, Jelle; Veenstra, Rense; Vries, J. de; Hoekstra, H.J.; Leij, Lou F. M. H. de; Vries, Elisabeth G.E. de

doi:10.1002/ijc.11295

Cited by 42 publications

(24 citation statements)

References 47 publications

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“…Our findings are in contrast to the recent results published by Schroder et al (2003) who found that in SNs immunostaining appears to be more sensitive/specific than quantitative PCR for breast tumour cell detection. The apparent discrepancy between our results and that of Schroder et al are likely due to the increased sensitivity achieved by the use of a multimarker panel in combination with the QDA score.…”

Section: Discussioncontrasting

confidence: 99%

Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple mRNA markers

et al. 2004

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Disseminated breast tumour cells in sentinel lymph nodes (SNs) were evaluated by quantitative real-time PCR and the sensitivity of this assay was compared to the routine histological analysis. First, several candidate marker genes were tested for their specificity in axillary lymph nodes (ALN) of 50 breast cancer patients and 43 women without breast cancer. The marker gene panel selected, designed to detect the mRNA of CK19, p1B, EGP2 and SBEM, was subsequently applied to detect metastases in 70 SNs that were free of metastases as determined by standard histological evaluation. Remarkably, seven negative SNs showed increased marker gene expression, suggesting the presence of (micro) metastases. Four of these seven SNs positive by real-time PCR proved to contain tumour deposits after careful review of the slides or further sectioning of the paraffin-embedded material. In three PCR positive SNs, however, no tumour cells were found by haematoxylin and eosin staining (H&E) and immunohistologically analysis. The quantitative real-time PCR assay with multiple mRNA markers for the detection of disseminated breast cancer cells in SNs thus resulted in an upstaging of SNs containing metastastic disease of 10% compared to the routine histological analysis. The application of this technique may be of clinical relevance, as it is suggested that micrometastatic disease in SNs are associated with further nodal non-SN metastases in breast cancer.

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Section: Discussioncontrasting

confidence: 99%

Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple mRNA markers

et al. 2004

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“…TACSTD1, also known as EPCAM, EGP-2, KS1/4, GA-733-2, and MIC-18, is a human cell surface antigen that is defined by the monoclonal antibody AUAI. TACSTD1 is also the antigen for the antibody Ber-Ep4 and as such has been used in studies to detect lymph node micrometastases in a variety of tumor types, including SCCHN (41,49,50). A recent study has also shown that TACSTD1 is a good marker for RT-PCR-based identification of lymph node micrometastasis in non-small cell lung cancer (9).…”

Section: Discussionmentioning

confidence: 99%

Molecular Staging of Cervical Lymph Nodes in Squamous Cell Carcinoma of the Head and Neck

Ferris

Raja³

et al. 2005

Cancer Research

101

119

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Clinical staging of cervical lymph nodes from patients with squamous cell carcinoma of the head and neck (SCCHN) has only 50% accuracy compared with definitive pathologic assessment. Consequently, both clinically positive and clinically negative patients frequently undergo neck dissections that may not be necessary. To address this potential overtreatment, sentinel lymph node (SLN) biopsy is currently being evaluated to provide better staging of the neck. However, to fully realize the potential improvement in patient care afforded by the SLN procedure, a rapid and accurate SLN analysis is necessary. We used quantitative reverse transcription--PCR (QRT-PCR) to screen 40 potential markers for their ability to detect SCCHN metastases to cervical lymph nodes. Seven markers were identified with good characteristics for identifying metastatic disease, and these were validated using a set of 26 primary tumors, 19 histologically positive lymph nodes, and 21 benign nodes from patients without cancer. Four markers discriminated between positive and benign nodes with accuracy > >97% but only one marker, pemphigus vulgaris antigen (PVA), discriminated with 100% accuracy in both the observed data and a statistical bootstrap analysis. A rapid QRT-PCR assay for PVA was then developed and incorporated into a prototype instrument capable of performing fully automated RNA isolation and QRT-PCR. The automated analysis with PVA provided perfect discrimination between histologically positive and benign lymph nodes and correctly identified two lymph nodes with micrometastatic tumor deposits. These assays were completed ( from tissue to result) in f f30 minutes, thus demonstrating the feasibility of intraoperative staging of SCCHN SLNs by QRT-PCR. (Cancer Res 2005; 65(6): 2147-56)

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“…CK19 is one of the keratins targeted by the AE1/AE3 antibody cocktail and, as such, is frequently used by pathologists for immunohistochemical detection or confirmation of lymph node metastases in many cancer types. CK19 has also been used in many RT-PCR-based studies to detect occult lymph node disease in breast cancer (21,24,25). Unfortunately, this gene has at least two pseudogenes that make it difficult to develop a cDNA-specific assay.…”

Section: Discussionmentioning

confidence: 99%

“…TACSTD1 is also known as EPCAM, EGP-2, KS1/4, GA-733-2, and MIC-18 and encodes a cell surface antigen that is defined by the monoclonal antibody AUAI. TACSTD1 is also the antigen for the antibody Ber-Ep4 and as such has been used in several studies to detect lymph node micrometastases in a variety of tumor types, including NSCLC (8,25,26). The recent studies by Wallace et al (22) and Mitas et al (23) also found TACSTD1 (KS1/4 in their article) to be a good RT-PCR-based marker for identification of lymph node metastasis in mediastinal lymph nodes from NSCLC patients.…”

Section: Discussionmentioning

confidence: 99%

A Combination of Molecular Markers Accurately Detects Lymph Node Metastasis in Non–Small Cell Lung Cancer Patients

Xi¹,

Coello²,

Litle³

et al. 2006

Clinical Cancer Research

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Occult lymph node metastasis (micrometastasis) is a good prognostic indicator in non^small cell lung cancer (NSCLC) and could be used to direct adjuvant chemotherapy in stage I patients. This study was designed to evaluate molecular markers for detection of occult lymph node metastasis in NSCLC, define the best marker or marker combination to distinguish positive from benign lymph nodes, and evaluate these markers in lymph nodes from pathologically node-negative (pN 0 ) NSCLC patients. Potential markers were identified through literature and database searches and all markers were analyzed by quantitative reverse transcription-PCR in a primary screen of six NSCLC specimens and 10 benign nodes. Selected markers were further evaluated on 21primary NSCLC specimens, 21positive nodes, and 21benign nodes, and the best individual markers and combinations were identified. A combination of three markers was further validated on an independent set of 32 benign lymph nodes, 38 histologically positive lymph nodes, and 462 lymph nodes from 68 pN 0 NSCLC patients. Forty-two markers were evaluated in the primary screen and eight promising markers were selected for further analysis. A combination of three markers (SFTPB, TACSTD1, and PVA) was identified that provided perfect classification of benign and positive nodes in all sample sets. PVA and SFTPB are particularly powerful in tumors of squamous and adenocarcinoma histologies, respectively, whereas TACSTD1 is a good general marker for NSCLC metastasis.The combination of these genes identified 32 of 462 (7%) lymph nodes from 20 of 68 (29%) patients as potentially positive for occult metastasis. Long-term follow-up will determine the clinical relevance of these findings.

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Detection of micrometastatic breast cancer by means of real time quantitative RT‐PCR and immunostaining in perioperative blood samples and sentinel nodes

Abstract: The aim of our study was to detect micrometastatic breast cancer by epithelial glycoprotein-2 (EGP-2) and cytokeratin 19 (CK19), using immunostaining and real time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).

Cited by 42 publications

References 47 publications

Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple mRNA markers

Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple mRNA markers

Molecular Staging of Cervical Lymph Nodes in Squamous Cell Carcinoma of the Head and Neck

A Combination of Molecular Markers Accurately Detects Lymph Node Metastasis in Non–Small Cell Lung Cancer Patients

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