Detection of circulating tumor cells and tumor stem cells in patients with breast cancer by using flow cytometry

Circulating tumor stem cells (CTSC), a subpopulation of circulating tumor cells (CTC), may lead to recurrent diseases. The aim of this study was to detect CTC (CD45(-)EpCAM(+)) and CTSC (CD45(-)EpCAM(+)CD44(+)CD24(-)) of breast cancer (BC) patients, as well as to explore their clinical relevance. CTC and CTSC in peripheral blood (PB) of 45 female BC patients were detected by using flow cytometry (FCM). SKBR-3 cells were mixed with MNC of four healthy volunteers at different ratios in order to evaluate the sensitivity of FCM. Real-time quantitative polymerase chain reaction (QRT-PCR) was conducted and compared with FCM. The expression of EPCAM between CTC < 50 and CTC a parts per thousand yenaEuro parts per thousand 50 groups (19.98 +/- 23.93 versus 29.46 +/- 29.27 x 10(-5)), and the expression of CD44 between CTSC negative and positive groups (0.85 +/- 0.91 versus 0.81 +/- 0.75) were statistically the same. FCM had higher specificity than QRT-PCR. Statistical differences were obtained between CTC < 50 and CTC a parts per thousand yenaEuro parts per thousand 50 groups among different TNM stages, histology stages, estrogen receptor (ER) status and progesterone receptor (PR) status (P < 0.05). Statistical differences between CTSC negative and positive groups within different TNM stages and regional lymph node metastasis (RLNM) status (P < 0.05) were also obtained. Moreover, the percentage of CTC on CD45 negative cells (CD45(-)C) among different clinical pathology was statistically different, P = 0.000. Additionally, the percentage of CTSC on CD45(-)C with TNM stage was rising (0: 0.00 +/- 0.00aEuro degrees, I: 0.03 +/- 0.05aEuro degrees, II: 0.06 +/- 0.14aEuro degrees, III: 0.10 +/- 0.09aEuro degrees, IV: 0.29 +/- 0.35aEuro degrees, P = 0.034). Statistical difference in the percentage of CTSC on CD45(-)C among different RLNM status (P = 0.001) was also obtained. FCM to detect CTC and CTSC may be used to diagnose disease at early stage, to guide clinical therapy or to predict prognosis.