CD24-low Cancer stem-cell like cell fraction in Notch inhibited transformed human esophageal cells

Comparison of CD24-low/CD44-high (CD44H) and CD24-high/CD44-low (CD44L) cell fractions in transformed human esophageal cells with Notch inhibition. CD44H cells display enhanced malignant potential (e.g. tumorigenicity, colony formation) compared to CD44L cells. NOTCH signaling was suppressed by retrovirus-mediated stable transduction of dominant-negative Mastermind-like1 (DNMAML1) in transformed human esophageal keratinocyte cell line EPC2-T (EPC2-hTERT-EGFR-cyclin D1-p53R175H) as described (Gastroenterology. 2010;139:2113-23), resulting in creation of cell line designated 4801. 4801 cells were sorted by flow cytometry for a cell fraction expressing a low level of CD24 and a high level of CD44 (CD44H) as well as that expressing a high level of CD24 and a low level of CD44 (CD44L). The former (CD44H) displayed fibroblast-like mesenchymal morphology while the latter (CD44L) remained cuboidal epithelioid morphology in culture. CD24-low and CD24-high cells were subjected to RNA extraction and gene array experiments.