Reverse-transcription polymerase chain reaction of cytokeratin 19(CK19) in detection of circulating tumor cells in breast cancer / 第二军医大学学报

ABSTRACT

Objective:To evaluate the value of CK19 mRNA detection in diagnosis of breast cancer. Methods: Patients were categorized into 3 groupsGroup 1 consisting of 77 stageⅠ/Ⅱbreast cancer patients, group 2 consisting of 77 stage Ⅲ/Ⅳ breast cancer patients, and control group consisting of 40 non-cancer volunteers. Total RNA was extracted from peripheral blood cells with the RNA Kit as described by the manufacturer, which was followed by one step RT-PCR of RNA. Agarose gel electrophoresis and sequencing were carried out to confirm the amplification of expected sequence (314 bp). ?~2 test was used to compare CK19 mRNA positive rates in different stages of breast cancer. Results: 15/77 (19.48%) of stage Ⅰ/Ⅱ breast cancers patients, 31/77 (40.26%) of stage Ⅲ/Ⅳ breast cancers patients and 2/40 (5.00%) of non-cancer volunteers demonstrated positive RT-PCR results. The positive rate of CK19 mRNA in peripheral blood of stage Ⅰ/Ⅱ or stage Ⅲ/Ⅳ breast cancer patients was significantly higher than that of non-cancer volunteers(P=0.037, P=0.001). The positive rate of CK19 mRNA in peripheral blood of stage Ⅲ/Ⅳbreast cancer patients was significantly higher than that of stageⅠ/Ⅱbreast cancer patients(P=0.005). Conclusion: CK19 RT-PCR is a promising method for detecting circulating tumor cells in patients with breast cancer, which may provide valuable information for tumor staging and treatment.