Detection of disseminated pancreatic cells by amplification of cytokeratin-19 with quantitative RT-PCR in blood, bone marrow and peritoneal lavage of pancreatic carcinoma patients.
OBJECTIVE
To evaluate the diagnostic potential of cytokeratin-19 (CK-19) mRNA for the detection of disseminated tumor cells in blood, bone marrow and peritoneal lavage in patients with ductal adenocarcinoma of the pancreas.


METHODS
Sixty-eight patients with pancreatic cancer (n = 37), chronic pancreatitis (n = 16), and non-pancreatic benign surgical diseases (n = 15, control group) were included in the study. Venous blood was taken preoperatively, intraoperatively and at postoperative d 1 and 10. Preoperative bone marrow aspirates and peritoneal lavage taken before mobilization of the tumor were analyzed. All samples were evaluated for disseminated tumor cells by CK-19-specific nested-PCR and quantitative fluorogenic RT-PCR.


RESULTS
CK-19 mRNA expression was increased in 24 (64%) blood samples and 11 (30%) of the peritoneal lavage samples in the patients with pancreatic cancer. In 15 (40%) of the patients with pancreatic cancer, disseminated tumor cells were detected in venous blood and bone marrow and/or peritoneal lavage. In the peritoneal lavage, the detection rates were correlated with the tumor size and the tumor differentiation. CK-19 levels were increased in pT3/T4 and moderately/poorly differentiated tumors (G2/G3). Pancreatic cancer patients with at least one CK-19 mRNA-positive sample showed a trend towards shorter survival. Pancreatic cancer patients showed significantly increased detection rates of disseminated tumor cells in blood and peritoneal lavage compared to the controls and the patients with chronic pancreatitis.


CONCLUSIONS
Disseminated tumor cells can be detected in patients with pancreatic ductal adenocarcinoma by CK-19 fluorogenic RT-PCR. In peritoneal lavage, detection rate is correlated with tumor stage and differentiation. In the clinical use, CK-19 is suitable for the distinction between malignant and benign pancreatic disease in combination with other tumor-specific markers.
Hoffmann
Katrin
K
Department of Visceral, Transplantation, Thoracic and Vascular Surgery, University of Leipzig, Germany. katrin.hoffmann@med.uni-heidelberg.de
Kerner
Christiane
C
Wilfert
Wolfgang
W
Mueller
Marc
M
Thiery
Joachim
J
Hauss
Johann
J
Witzigmann
Helmut
H
D014408:Biomarkers, Tumor / Q000032:analysis* / Q000097:blood; D001853:Bone Marrow / Q000473:pathology; D021441:Carcinoma, Pancreatic Ductal / Q000175:diagnosis* / Q000473:pathology; D006801:Humans; D053539:Keratin-19 / Q000235:genetics*; D009360:Neoplastic Cells, Circulating / Q000737:chemistry* / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000175:diagnosis* / Q000473:pathology; D010533:Peritoneal Lavage; D012333:RNA, Messenger / Q000032:analysis / Q000097:blood; D012334:RNA, Neoplasm / Q000032:analysis* / Q000097:blood; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D014408:Biomarkers, Tumor; D053539:Keratin-19; D012333:RNA, Messenger; D012334:RNA, Neoplasm