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The invasive, mesenchymal phenotype of CD44 pos CD24 neg  breast cancer cells has made them a promising target for eliminating the metastatic capacity of primary tumors. It has been previously demonstrated that CD44 neg/low CD24 pos  breast cancer cells lack the ability to give rise to their invasive CD44 pos CD24 neg  counterpart. Here we demonstrate that noninvasive, epithelial-like CD44 pos CD24 pos  cells readily give rise to invasive, mesenchymal CD44 pos CD24 neg  progeny  in vivo  and  in vitro . This interconversion was found to be dependent upon Activin/Nodal signaling.    Methods  Breast cancer cell lines were sorted into CD44 pos CD24 pos  and CD44 pos CD24 neg  populations to evaluate their progeny for the expression of CD44, CD24, and markers of a mesenchymal phenotype. The populations, separated by fluorescence activated cell sorting (FACS) were injected into immunocompromised mice to evaluate their tumorigenicity and invasiveness of the resulting xenografts.    Results  CD24 expression was dynamically regulated  in vitro  in all evaluated breast cancer cell lines. Furthermore, a single noninvasive, epithelial-like CD44 pos CD24 pos  cell had the ability to give rise to invasive, mesenchymal CD44 pos CD24 neg  progeny. Importantly, this interconversion occurred  in vivo  as CD44 pos CD24 pos  cells gave rise to xenografts with locally invasive borders as seen in xenografts initiated with CD44 pos CD24 neg  cells. Lastly, the ability of CD44 pos CD24 pos  cells to give rise to mesenchymal progeny, and  vice versa , was blocked upon ablation of Activin/Nodal signaling.    Conclusions  Our data demonstrate that the invasive, mesenchymal CD44 pos CD24 neg  phenotype is under dynamic control in breast cancer cell lines both  in vitro  and  in vivo . Furthermore, our observations suggest that therapies targeting CD44 pos CD24 neg  tumor cells may have limited success in preventing primary tumor metastasis unless Activin/Nodal signaling is arrested.

Dynamic regulation of CD24 and the invasive, CD44posCD24negphenotype in breast cancer cell lines