The detection of circulating tumor cells of breast cancer patients by using multimarker (Survivin, hTERT and hMAM) quantitative real-time PCR.
OBJECTIVE
To develop a specific, reliable assay for detecting circulating tumor cells (CTC) in peripheral blood of breast cancer patients.


METHODS
94 breast cancer patients, 35 patients with benign breast tumor, 40 healthy individuals, and 25 patients with other solid tumors were evaluated by quantitative real-time reverse transcription-PCR (qRT-PCR) for detecting Survivin, hTERT, and hMAM mRNA in peripheral blood (PB) of breast cancer patients. In addition, analyses were carried out for their correlation with patients' clinicopathologic features.


RESULTS
The sensitivity of Survivin, hTERT, and hMAM mRNA in the PB of breast cancer patients was 36.2%, 59.6% and 33.0%, respectively. Survivin and hTERT were detected in the PB patients with solid tumors other than breast cancer, but hMAM mRNA was only detected in breast cancer patients. The sensitivity of three combined markers in the parallel test was 70.2%.Compared to that of single marker detection, the three combined markers' percentage was significantly higher. However, the specificity of three combined markers of serial test was 100%. The expression of the three mRNAs significantly correlated with TNM stage, and lymph node metastasis.


CONCLUSIONS
Survivin, hTERT and hMAM mRNA assays are powerful methods for detection of CTC of breast cancer patients. With combination of the three markers for detection of CTC of breast cancer, the parallel test increases the sensitivity. This analysis can offer a simple, noninvasive, and promising adjuvant tool for the early detection of micrometastatic tumor cells in breast cancer patients.
Shen
ChangXin
C
Clinical Laboratory Medicine Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Hu
LiHua
L
Xia
Lin
L
Li
YiRong
Y
D001943:Breast Neoplasms / Q000175:diagnosis* / Q000473:pathology; D016022:Case-Control Studies; D004268:DNA-Binding Proteins / Q000235:genetics; D005260:Female; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics; D009362:Neoplasm Metastasis / Q000175:diagnosis*; D009360:Neoplastic Cells, Circulating; D012334:RNA, Neoplasm / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D000077022:Survivin; D019098:Telomerase / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors
C107489:BIRC5 protein, human; D004268:DNA-Binding Proteins; D051034:Inhibitor of Apoptosis Proteins; C430872:MAML1 protein, human; D008869:Microtubule-Associated Proteins; D012334:RNA, Neoplasm; D000077022:Survivin; D015534:Trans-Activators; D014157:Transcription Factors; C509186:TERT protein, human; D019098:Telomerase