title: Epithelial-mesenchymal transition and stem cell markers in patients with HER2-positive metastatic breast cancer.
abstract: 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(+) metastatic breast cancer (MBC). Epithelial cells were enriched from peripheral blood mononuclear cells (PBMC) using antibody-coated anti-CD326 antibody (CD326(+)) magnetic beads, and the residual CD326(-) PBMCs were further depleted of leukocytes using anti-CD45 antibody-coated magnetic beads (CD326(-)CD45(-)). RNA was extracted from all cell fractions, reverse transcribed to cDNA, and subjected to quantitative reverse transcription PCR to detect EMT-TFs (TWIST1, SNAIL1, ZEB1, and TG2) 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(-)CD45(-) cell fraction of 60.7% of patients. The CD326(-)CD45(-) fraction of patients with elevated SNAIL1 and ZEB1 transcripts also had a higher percentage of ALDH(+)/CD133(+) cells in their blood than did patients with normal SNAIL1 and ZEB1 expression (P = 0.038). Our data indicate that patients with HER2(+) MBCs have EMT-CTCs. Moreover, an enrichment of CSCs was found in CD326(-)CD45(-) cells. Additional studies are needed to determine whether EMT-CTCs and CSCs have prognostic value in patients with HER2(+) MBCs treated with trastuzumab-based therapy.
authors.affiliation: Department of Hematopathology, Unit 0054, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Blvd., 77030, Houston, TX, USA.
authors.forename: Antonio
authors.initials: A
authors.lastname: Giordano
authors.forename: Hui
authors.initials: H
authors.lastname: Gao
authors.forename: Simone
authors.initials: S
authors.lastname: Anfossi
authors.forename: Evan
authors.initials: E
authors.lastname: Cohen
authors.forename: Michal
authors.initials: M
authors.lastname: Mego
authors.forename: Bang-Ning
authors.initials: BN
authors.lastname: Lee
authors.forename: Sanda
authors.initials: S
authors.lastname: Tin
authors.forename: Michele
authors.initials: M
authors.lastname: De Laurentiis
authors.forename: Charla A
authors.initials: CA
authors.lastname: Parker
authors.forename: Ricardo H
authors.initials: RH
authors.lastname: Alvarez
authors.forename: Vicente
authors.initials: V
authors.lastname: Valero
authors.forename: Naoto T
authors.initials: NT
authors.lastname: Ueno
authors.forename: Sabino
authors.initials: S
authors.lastname: De Placido
authors.forename: Sendurai A
authors.initials: SA
authors.lastname: Mani
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: Esteva
authors.forename: Massimo
authors.initials: M
authors.lastname: Cristofanilli
authors.forename: James M
authors.initials: JM
authors.lastname: Reuben
mesh_terms: D000328:Adult; D000368:Aged; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001943:Breast Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002452:Cell Count; D045744:Cell Line, Tumor; D018572:Disease-Free Survival; D000071858:Epithelial Cell Adhesion Molecule; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D053208:Kaplan-Meier Estimate; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D016896:Treatment Outcome
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D017493:Leukocyte Common Antigens; D009363:Neoplasm Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human