The detection of disseminated tumor cells in bone marrow from colorectal-cancer patients by a cytokeratin-20-specific nested reverse-transcriptase-polymerase-chain reaction is related to the stage of disease.
The dissemination of cancer cells is a prerequisite in the development of micrometastases and solid metastases. Our previous examinations of these cells were based on immunocytological staining of tumor-associated antigens and cytokeratins. We have now developed a highly sensitive and specific detection method based on a nested reverse-transcriptase-polymerase-chain reaction (RT-PCR) of cytokeratin-20 (CK-20) mRNA. Using this method, we examined the bone marrow of 57 patients with colorectal cancer and detected increasing numbers of CK-20-positive samples, depending on the UICC stage. Some 35% of all bone-marrow samples tested positive for CK-20: none were found in colorectal cancer stage 1, 24% were in stage II, 31% in stage III and 71% in stage IV. Investigation of bone-marrow specimens of patients with pancreatic cancer showed that 4 out of 11 patients were positive for CK-20 mRNA. To confirm that sample positivity for CK-20 expression was due to disseminated tumor cells, we examined bone marrow from a control group (n = 16) without apparent carcinoma. In this group, 15 out of 16 donors were CK-20-negative, while one donor with familial adenomatous polyposis showed a CK-20-specific signal.
Soeth
E
E
Department of General Surgery and Thorax Surgery, University Hospital Kiel, Germany.
Röder
C
C
Juhl
H
H
Krüger
U
U
Kremer
B
B
Kalthoff
H
H
D001483:Base Sequence; D014408:Biomarkers, Tumor; D001853:Bone Marrow / Q000473:pathology*; D015179:Colorectal Neoplasms / Q000737:chemistry / Q000473:pathology*; D005770:Gastrointestinal Neoplasms / Q000737:chemistry / Q000473:pathology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics*; D053554:Keratin-20; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D009367:Neoplasm Staging; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D014407:Tumor Cells, Cultured
D014408:Biomarkers, Tumor; D007381:Intermediate Filament Proteins; C507499:KRT20 protein, human; D053554:Keratin-20; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase