Detection of circulating cancer cells by nested reverse transcription-polymerase chain reaction of cytokeratin-19 (K19)--possible clinical significance in advanced gastric cancer.

摘要: Intermediate filament cytokeratin-19 (K19) protein is expressed in normal and malignant gastrointestinal epithelial cells, but not peripheral blood (PB). Small amount of circulating gastric cancer cells can be detected by a sensitive nested reverse transcription-polymerase chain reaction (RTPCR) with primers specific for K19 mRNA. Thirty-four PB samples obtained from patients inoperable/metastatic were examined. The mononuclear cell (MNC) fraction was collected Ficoll centrifugation, followed total RNA extraction acid guanidinium thiocyanate-phenol-chloroform method. 8 lines the 33 healthy adults used as positive negative controls, respectively. DNA fragment 774 bp amplified internal found to highly reliable marker mRNA expression. sensitivity detection between 1 10 cells/10(6) MNCs. transcripts 20.6% (7/34; 8-37%, 95% C.I.) samples. None other pertinent clinicopathological features, including disease extent histopathologic types tumors, related expression PB. All 34 had been treated systemic chemotherapy. Among 17 non-responders chemotherapy, survival 4 detectable significantly shorter than that 13 without their (p = 0.014). However, impact less significant whose tumors responded Of whole group patients, median 7 27 3.5 months, respectively 0.368). We concluded detecting RT-PCR associated poor prognosis cancer, particularly those who are responsive