**Acknowledgements**

We successfully confirmed that the existence of CTSCs and also reveal the relationship between CTSCs level and different TNM stages. The correlation of CTSCs and clinical pathologic

cells was not correlated with clinical features such as lymph node status, tumor size, histology

were related with distant metastasis, particularly osseous [45]. And we found the expression of CTSCs was quite related with RLNM status. It was a novel way for treatment targeting at

Cancer stem cells (CSCs) had been confirmed existed in many kinds of epithelial malignancy [47]. And the CSCs were considered as a subpopulation of tumor cells [48]. The mutation of normal stem cells resulted in the genesis of CSCs, which had been demonstrated by Cariati and Purushotham [49]. Therefore, CTSCs were suggested that it was generated not only from CTCs but also from normal stem cells. Further researches needed to be performed to confirm

Detection of CTCs and CTSCs by using multiparameter flow cytometry was considered as an effective technique on monitoring disease development and evaluating prognosis [50, 51]. Using multiparameter flow cytometry to detect CTC and CTSC in peripheral blood may provide new opportunities for the early diagnosis of invasive breast cancer, guide us to select optimal therapeutic regimens and help us to predict the prognosis. Moreover, we wish to

CTSCs needed to be demonstrated and we have established a firm basis for following research.

Detection of CTCs by using multiparameter flow cytometry was effective and it has the potential to be a valuable method for both prognosis assessment and cancer research in breast

There is no doubt that CTCs would become an important test in clinical in near future. The challenge of the achievement contains the heterogeneity of CTCs and big database of clinical on CTCs. The heterogeneity of CTCs reflected in different tumor related proteins and mor‐ phology. However, researchers have pay more and more interesting on EMT phenotype that is common in all kinds of tumors. It will make CTCs detection accessible by decipherment of

CD24-

CD24-

ESA- expressing. Further researches on

CD24- tumor stem cells

cancer stem

features remains unclear before. Previous study had reported that the CD44+

It was reported that the patients who had high expression of CD44+

CTSCs to prevent metastasis and to evaluate the prognosis.

isolate the CTSCs that have the marker of CD44+

grade, ER, and PR, or HER‐2 [45, 46].

it.

208 Tumor Metastasis

**5. Conclusion**

**6. Looking forward**

conundrums underlying EMT phenotype.

cancer.

We thank the Department of oncology and cell therapy in Union Hospital for samples providing.
