Detection of telomerase activity in peritoneal lavage fluid from patients with gastric cancer using immunomagnetic beads.
Cytologic examination of peritoneal lavage fluid is a useful predictor of peritoneal recurrence in gastric cancer. However, this technique is not overly sensitive and requires special abilities in the cytologist. In this study, telomerase activity was used to detect free cancer cells in peritoneal lavage fluid from patients with gastric cancer. In the first part, 12 lavage-fluid samples obtained from 12 patients with gastric cancer were analysed using the conventional telomeric repeat amplification protocol (TRAP) assay. Three of five patients with early gastric cancer had positive telomerase activity. These false-positive results may have been due to lymphocyte contamination. Furthermore, polymerase chain reaction inhibitors were also detected in the lavage-fluid samples. Therefore, we developed a novel method for elimination of haematopoietic cell and Taq polymerase inhibitors to increase the accuracy of the TRAP assay using immunomagnetic beads, which bind to most normal and neoplastic human epithelial cells. Telomerase activity was found in 10 of 20 (50%) lavage-fluid samples from patients with serosal or subserosal invasion. Cytologic examination was positive in nine of 20 (45%) samples. Both the telomerase activity and cytology were negative in all 14 patients without serosal or subserosal invasion. These results suggest that the TRAP assay combined with immunomagnetic beads might be useful for detection of free cancer cells in the peritoneal space in gastric cancer without the aid of an experienced cytologist.
Mori
N
N
Departments of Surgery II, Biochemistry II, Yamaguchi University School of Medicine, 1-1-1 Minamikogushi, Ube, Yamaguchi, 755-8505, USA.
Oka
M
M
Hazama
S
S
Iizuka
N
N
Yamamoto
K
K
Yoshino
S
S
Tangoku
A
A
Noma
T
T
Hirose
K
K
D000230:Adenocarcinoma / Q000175:diagnosis* / Q000201:enzymology / Q000473:pathology; D000328:Adult; D000368:Aged; D001202:Ascitic Fluid / Q000201:enzymology*; D005189:False Positive Reactions; D005260:Female; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods; D008297:Male; D008875:Middle Aged; D010533:Peritoneal Lavage; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D013274:Stomach Neoplasms / Q000175:diagnosis* / Q000201:enzymology / Q000473:pathology; D019098:Telomerase / Q000032:analysis*; D014407:Tumor Cells, Cultured
D019098:Telomerase