Putative markers for the detection of breast carcinoma cells in blood

Abstract

Summary The aim of this study was to investigate certain genes for their suitability as molecular markers for detection of breast carcinoma cells using the reverse transcriptase-polymerase chain reaction (RT-PCR). RNA was prepared from MCF-7 breast carcinoma cells and peripheral blood leucocytes of healthy female volunteers. This RNA was screened for mRNA of MUC1, cytokeratin 19 (CK19) and CD44 (exons 8-11) by RT-PCR and the results validated by Southern blots. Variable degrees of expression of MUC1 and CD44 (exons 5-11) were detected in normal peripheral blood, rendering these genes non-specific for epithelial cells and therefore unsuitable for use as markers to detect breast carcinoma cells. Although CK1 9 mRNA was apparently specific, it was deemed unsuitable for use as a marker of breast cancer cells in light of its limited sensitivity. Furthermore, an attempt at using nested primers to increase sensitivity resulted in CK19 mRNA being detected after two amplification rounds in blood from healthy volunteers.

Analysis of potential markers for detection of submicroscopic lymph node metastases in breast cancer

“…Two previous reports have suggested MUC-1 to be tissue specific (Noguchi et al, 1994(Noguchi et al, , 1996a; however, there is no reported data on the number of control patients, and a total of only ten lymph nodes assessed. Our results concur with that of other groups in showing that MUC-1 is expressed in cells of lymphohaemopoetic origin (Hoon et al, 1995;Eltahir et al, 1998). Several studies in breast cancer have reported K19 to be a tissue-specific marker (Datta et al, 1994;Schoenfeld et al, 1994Schoenfeld et al, , 1996Schoenfeld et al, , 1997Noguchi et al, 1996a;Eltahir et al, 1998;Lockett et al, 1998).…”

“…However, data from studies assessing the detection of disease in colon and lung cancer have raised questions with regard to the tissue specificity of these markers (Adams et al, 1995;Burchill et al, 1995;Hoon et al, 1995;Krismann et al, 1995;Denis et al, 1997;Dingemans et al, 1997;Eltahir et al, 1998) CEA gene expression must be regarded with caution as a tumour specific marker, as it has been detected by nested RT-PCR in normal tissues. Jonas et al (1996) found 23% of controls without cancer had evidence of CEA expression in peripheral blood, which the authors propose may be due to venepuncture-induced skin contamination.…”

“…Whilst lacking specific markers expressed by breast cancer cells a number of research groups have used cytokeratins 18 and 19, epithelial mucin (MUC1), carcinoembryonic antigen (CEA), CD 44 and maspin as transcript markers for the detection of submicroscopic metastases in lymph nodes, bone marrow or peripheral whole blood by reverse transcription PCR (RT-PCR) (Matsumura and Tarin, 1992;Datta et al, 1994;Gerhard et al, 1994;Noguchi et al, 1994Noguchi et al, , 1996aNoguchi et al, , 1996bSchoenfeld et al, 1994Schoenfeld et al, , 1996Schoenfeld et al, , 1997Brown et al, 1995;Mori et al, 1995;Gunn et al, 1996;Luppi et al, 1996;Yun et al, 1997;Eltahir et al, 1998;Lockett et al, 1998). However, there appears to be conflicting data regarding the specificity of some of these cell type specific markers in particular MUC-1 (Noguchi et al, 1994;Hoon et al, 1995), CD44 (Matsumura and Tarin, 1992;Eltahir et al, 1998) and K19 (Traweek et al, 1993;Schoenfeld et al, 1994Schoenfeld et al, , 1996Burchill et al, 1995;Krismann et al, 1995;Gunn et al, 1996;Dingemans et al, 1997;Yun et al, 1997;Eltahir et al, 1998). Maspin expression has been previously reported as being a specific marker for breast cancer (Luppi et al, 1996); however, there have been no larger confirmatory studies to assess the accuracy and reproducibility of these findings.…”

High-sensitivity array analysis of gene expression for the early detection of disseminated breast tumor cells in peripheral blood

“…muc-1 and her2͞neu have also been reported as useful markers for breast cancer in some blood studies (4, 18) but not others (19,20). Our results found muc-1 high in normal blood and hence not a useful marker (Fig.…”

Mammaglobin Gene Expression: A Superior Marker of Breast Cancer Cells in Peripheral Blood in Comparison to Epidermal-Growth-Factor Receptor and Cytokeratin-19

“…This high rate of false positivity might be caused primarily by the high sensitivity of the nested RT-PCR assay picking up illegitimate low-level expression of CK-19 in mononuclear blood cells (Chelly et al, 1989;Eltahir et al, 1998;Schoenfeld et al, 1994). So far, EGF-R-specific transcripts have not been detected in human blood mononuclear cells.…”