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

doi:10.3877/cma.j.issn.1673-5250.2006.06.103

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

图1　质粒(PCR4-T0P0 CK₁₉)实时定量PCR的扩增曲线(A)和标准曲线。(A)质粒的浓度自4 × 10⁵拷贝/ μl至40拷贝/ μl按10倍稀释的PCR曲线。(B)循环阈值与质粒浓度对数关系的标准曲线

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