Real-time quantitative RT-PCR detection of lymph node micrometastases of patients with endometrial cancer

Objective To develop a quantitative method for the detection of lymph node micrometastases of patients with endometrial cancer. Methods 218 lymph nodes from 14 patients were studied.Real-time quantitative analysis was performed using cytokeratin 19(CK19)as a gene marker. Results This method resulted in the reproducible quantification of 10 to 106 MCF-7 cells per 106 peripheral blood nucleated cells.CK19 mRNA was detected in 62 lymph node from 10 patients.33 lymph node from 6 patients were PCR positive.The PCR-positive rate of lymph node was higher in patients with pathologically metastatic lymph node than in patients with path-negative but PCR-positive lymph node.Furthermore, the CK19 mRNA background expression rate was higher in the lymph node of path-negative but PCR-positive patients than in lymph nodes of path-negative and PCR-negative patients. Conclusions Real-time quantitative RT-PCR with TaqMan probes is a sensitive, specific and rapid method for the detection of micrometastases in lymph node of patients with endometrial cancer. Key words: endometrial cancer; polymerase chain reaction; micrometastasis; cytokeratin