Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow.

Zippelius, Alfred; Kufer, Peter; Honold, G.; Köllermann, M. W.; Oberneder, R.; Schlimok, Günter; Riethmüller, Gert; Pantel, Klaus

doi:10.1200/jco.1997.15.7.2701

Cited by 264 publications

(150 citation statements)

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“…[39][40][41][42][43] The rate After chemotherapy and use of growth factors, peripheral blood stem cells can be processed and harvested in large of false-positive findings can be lowered by decreasing the number of PCR cycles and by using fewer cells, but this amounts. Chemotherapy followed by growth factor administration may mobilize tumor cells into peripheral blood.…”

Section: Preparation and Immunostaining Of Bone Marrow Specimens And mentioning

confidence: 99%

Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients

Schulze

Wischnik

et al. 1997

Bone Marrow Transplant

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Summary:therapy and stem cell support appears possible. Peters et al 2,3 reported a 78% overall survival (OS) in this high-risk breast cancer subgroup after high-dose chemotherapy folTwenty-one high-risk patients with primary stage II/III breast cancer were treated with high-dose chemolowed by stem cell support (median follow-up 4.5 years).Gianni et al 4 showed a OS of 78% after 5 years in these therapy comprising etoposide, ifosfamide, carboplatin and epirubicin (VIC-E). Tumor cells of epithelial origin patients. Several authors have suggested that tumour cells present in stem cell transplants may contribute to relapse were analyzed using the monoclonal antibodies CK2 (IgG 1 ) and A45-B/B3 (IgG 1 ) against cytokeratin (CK) in patients treated with high-dose chemotherapy and stem cell support. [5][6][7][8][9] To determine the significance of tumor cell For the present study 21 female patients with high-risk patient number only a trend towards a superior relapsebreast cancer and involvement of seven to nine (three free survival in the patient group with CK negative patients) and 10 or more (18 patients) axillary lymph nodes transplants can be shown by Kaplan-Meier analysis.were used. Informed consent was obtained from all Keywords: tumor cells; peripheral blood stem cell transpatients. All patients had histologically proven resectable plantation; bone marrow; breast cancer breast cancer (19 invasive ductal, two invasive lobular).After an extensive diagnostic program, patients were staged according to the UICC breast cancer classification as folBreast cancer accounts for approximately 30% of all canlows: UICC stage IIA n = 4 (19%), IIB n = 8 (38%), IIIA cers in women. It is one of the most common malignancies n = 5 (24%), IIIB n = 4 (19%). Median age of the patients and the second leading cause of death in women. 1 The 10-at time of primary surgery was 50.8 years . No year relapse rate, increasing with the number of lymph patient had received previous chemotherapy, radiotherapy nodes involved, is about 20% for negative lymph node or immunotherapy. Patient characteristics are shown in stages and greater than 60% in women with one to three Table 1. lymph nodes involved. Patients with stage II/III breast cancer and 10 or more involved axillary lymph nodes can curTreatment rently expect only a 15-20% 10-year disease-free survival. Improvement of treatment outcome with standard-dose Patients were treated with two cycles of induction chemoinduction chemotherapy followed by high-dose chemotherapy with the standard-dose VIP-E regimen, which consisted of etoposide 500 mg/m 2 , ifosfamide 4000 mg/m 2 , cisplatin 50 mg/m 2 and epirubicin 50 mg/m 2 . 13 Mesna uro-

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Section: Preparation and Immunostaining Of Bone Marrow Specimens And mentioning

confidence: 99%

Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients

Schulze

Wischnik

et al. 1997

Bone Marrow Transplant

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“…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). However, there are also reports on the limitations of this method (Zippelius et al, 1997;Bostick et al, 1998a, b;Ko et al, 2000). The major limitation is related to the specificity in the detection of metastatic cancer cells.…”

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confidence: 99%

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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“…Expression of epithelial markers in the bone marrow of patients with nonepithelial tumours or inflammatory diseases has previously been reported (Zippelius et al, 1997;Zach et al, 1999;Ballestrero et al, 2001;Krüger et al, 2001). Therefore, we evaluated in normal blood cells the possibility of a cytokine-induced expression of maspin and MAM.…”

Section: Discussionmentioning

confidence: 93%

“…However, there are several factors such as pseudogenes, background gene expression and aberrant expression of epithelial genes in mesenchymal cells under several physiological or physiopathological conditions that can reduce the specificity of the assay and give rise to an unacceptable incidence of falsepositive results (Zippelius et al, 1997;Jung et al, 1998Jung et al, , 1999Ruud et al, 1999;Krüger et al, 2001;Jiang et al, 2002). Mammaglobin (MAM) and maspin are markers of major interest because some evidence indicates that they have a stronger relationship with breast tissue than several other epithelial markers such as cytokeratins, CEA and MUC-1 (Luppi et al, 1996;Zach et al, 1999;Grünewald et al, 2000;Ballestrero et al, 2001;Corradini et al, 2001;Silva et al, 2002;Zehentner and Carter, 2004).…”

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Effect of different cytokines on mammaglobin and maspin gene expression in normal leukocytes: possible relevance to the assays for the detection of micrometastatic breast cancer

et al. 2005

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In cancer patients, the ability to detect disseminated tumour cells in peripheral blood or bone marrow could improve prognosis and consent both early detection of metastatic disease and monitoring of the efficacy of systemic therapy. These objectives remain elusive mainly due to the lack of specific genetic markers for solid tumours. The use of surrogate tissue-specific markers can reduce the specificity of the assays and give rise to a clinically unacceptable false-positive rate. Mammaglobin (MAM) and maspin are two putative breast tissue-specific markers frequently used for detection of occult tumour cells in the peripheral blood, bone marrow and lymph nodes of breast cancer patients. In this study, it was evaluated whether MAM and maspin gene expression may be induced in the normal blood and bone marrow cells exposed to a panel of cytokines, including chemotactic factors (C5a, interleukin (IL)-8), LPS, proinflammatory cytokines (TNF-a, IL-1b) and growth factors (IL-3, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor). The experimental data show that all cytokines included in the panel, except for IL-8, were able to induce maspin expression; on the contrary, MAM gene was never induced. These results suggest that MAM is more specific than maspin and that the possible interference of cytokines should be taken into account in interpreting molecular assays for detection of isolated tumour cells.

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Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow.

Abstract: Limiting factors in the detection of micrometastatic tumor cells by RT-PCR are (1) the illegitimate transcription of tumor-associated or epithelial-specific genes in hematopoietic cells, and (2) the deficient expression of the marker gene in micrometastatic tumor cells.

Cited by 264 publications

References 28 publications

Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients

Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients

Quantitative evaluation of metastases in axillary lymph nodes of breast cancer

Effect of different cytokines on mammaglobin and maspin gene expression in normal leukocytes: possible relevance to the assays for the detection of micrometastatic breast cancer

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