Epithelial tumor cells in bone marrow of patients with colorectal cancer: immunocytochemical detection, phenotypic characterization, and prognostic significance.

Abstract

A monoclonal antibody (mAb) directed against the cytokeratin (CK) polypeptide no. 18 specifically expressed in cells derived from simple epithelia was used to detect epithelial tumor cells in bone marrow aspirates. Of 156 patients with colorectal carcinoma, 42 presented with cells at the time of primary surgery. The incidence of positive findings varied considerably with the size and the localization of the primary tumor, the involvement of regional lymph nodes, and the presence of clinically manifest metastases. Applying a sensitive double-staining procedure, we could demonstrate that epithelial cells in bone marrow showed a heterogeneic expression of receptors for epidermal growth factor (EGF-R) and transferrin (Tf-R) as well as of the proliferation-associated Ki67 antigen. Also human leukocyte antigen (HLA) class I antigens differed widely in their expression on the CK-positive cells. Clinical follow-up studies on 85 patients showed a significantly higher relapse rate in patients presenting with CK-positive cells in their bone marrow at the time of primary surgery. Twenty-three patients were monitored for the presence or absence of CK-positive cells in bone marrow over time. The majority of monitored patients (18 of 23) exhibited a constant pattern of immunocytochemical findings during the time of observation. Thus, the technique may be useful in identifying high-risk patients as well as in monitoring adjuvant therapeutic trials.

Tumor cell dissemination in colon cancer does not predict extrahepatic recurrence in patients undergoing surgery for hepatic metastases

“…The detection rate of 55% for CK + BM cells seems to be high at first view when compared to other published studies using immunocytochemistry for the detection of CK + cells, but this discrepancy can be explained by the number of cells analyzed. While Schlimok et al screened 1.5x10 5 BM cells resulting in a detection rate of 26.9% CK + cells, and Lindemann et al found 31.9% BM samples to be CK + when 3x10 5 cells were analyzed, we used more than 1x10 7 BM cells for the screening of CK + cells. The differences in the detection rate of CK + cells during surgery may be due to the different concepts of evaluation in each study.…”

“…Since this tumor cell spread is regularly underestimated by currently available clinical and pathological staging procedures, sensitive immunohistochemical and molecular techniques were developed to facilitate the detection of isolated carcinoma cells in blood, bone marrow (BM) or lymph nodes using monoclonal antibodies (Mab) against epithelium-specific proteins (3,4). CK + cells were detected in the BM of patients with colorectal cancer, and their presence has been associated with a significantly higher relapse rate (5,6). Furthermore, CK-20 RT-PCR transcripts in peritumoral histopathologic tumor-free (pN0) lymph nodes of colorectal cancer patients were shown to be an independent prognostic factor for overall survival (7,8).…”

Molecular cytogenetic analysis of cytokeratin 20-labeled cells in primary tumors and bone marrow aspirates from colorectal carcinoma patients

“…Tumor biopsies (1 -2 gm) were obtained during surgery and placed in phosphatecells in bone marrow aspirates will develop recurrent disease at distant sites (false-negatives), and 30% of buffered saline (PBS) (0.14 M NaCl, 2.7 mM KCl, 5.4 mM Na 2 HPO 4 , and 1.8 mM KH 2 PO 4 ; pH 7.0) on ice patients with CK / cells may not recur (false-positives). 7,8 Typically, 10 5 -10 6 bone marrow cells are prior to processing. Biopsies were sliced with a razor blade and lightly touched (''touch preps'') onto ethscreened for the presence of CK / cells.…”

“…Approximately 10 4 low density cells were deposited onto ethanol-cleaned glass vival. 6,7 The predictive value of micrometastases assays to stratify patients into groups at increased risk of deslides using a Shandon cytocentrifuge (Shandon, Pittsburgh, PA) and stored at 020 ЊC prior to immuveloping overt metastases is nonuniform. Approximately 40% of patients without detectable epithelial nostaining and FISH.…”

Biological therapy: approaches in colorectal cancer

“…Recent novel immunocytochemical techniques using immunobead polymerase chain reaction (PCR) for detection of the tumour-associated antigen carcinoembryonic antigen (CEA) have permitted the identification of single malignant cells in peripheral blood samples, bone marrow aspirates and peripheral stem cell harvests through the recognition of unique hybrid gene transcripts. (Schlimok et al, 1990;Lindeman et al, 1992;Hardingham et al, 1993;Johnson et al, 1995).…”