Abstract
<div>Abstract<p>Currently, there is extensive information about circulating tumor cells (CTC) and their prognostic value; however, little is known about other characteristics of these cells. In this prospective study, we assessed the gene transcripts of epithelial-to-mesenchymal transition—inducing transcription factors (EMT-TF) and cancer stem cell (CSC) features in patients with HER2<sup>+</sup> metastatic breast cancer (MBC). Epithelial cells were enriched from peripheral blood mononuclear cells (PBMC) using antibody-coated anti-CD326 antibody (CD326<sup>+</sup>) magnetic beads, and the residual CD326<sup>−</sup> PBMCs were further depleted of leukocytes using anti-CD45 antibody-coated magnetic beads (CD326<sup>−</sup>CD45<sup>−</sup>). RNA was extracted from all cell fractions, reverse transcribed to cDNA, and subjected to quantitative reverse transcription PCR to detect EMT-TFs (<i>TWIST1</i>, <i>SNAIL1</i>, <i>ZEB1</i>, and <i>TG2</i>) as a measure of CTCs undergoing EMT (EMT-CTCs). In addition, PBMCs were analyzed using multiparameter flow cytometry for ALDH activity and CSCs that express CD24, CD44, and CD133. Twenty-eight patients were included in this study. At least one EMT-TF mRNA was elevated in the CTCs of 88.2% of patients and in the CD326<sup>−</sup>CD45<sup>−</sup> cell fraction of 60.7% of patients. The CD326<sup>−</sup>CD45<sup>−</sup> fraction of patients with elevated <i>SNAIL1</i> and <i>ZEB1</i> transcripts also had a higher percentage of ALDH<sup>+</sup>/CD133<sup>+</sup> cells in their blood than did patients with normal <i>SNAIL1</i> and <i>ZEB1</i> expression (<i>P</i> = 0.038). Our data indicate that patients with HER2<sup>+</sup> MBCs have EMT-CTCs. Moreover, an enrichment of CSCs was found in CD326<sup>−</sup>CD45<sup>−</sup> cells. Additional studies are needed to determine whether EMT-CTCs and CSCs have prognostic value in patients with HER2<sup>+</sup> MBCs treated with trastuzumab-based therapy. <i>Mol Cancer Ther; 11(11); 2526–34. ©2012 AACR</i>.</p></div>
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