Figure 1.

NANOG expression positively correlates with cell proliferation, migration, and cancer stem-like properties in ESCC. (A) Western blotting was performed to analyze the ectopic expression of NANOG in ESCC. The protein concentrations of the samples were detected using the BCA assay. (B) Cell numbers after using a series of doses of NANOG expression in Eca-109 cells. Bottom panel: bright-field microscopy of cells stained with crystal violet solution on day 5 after NANOG overexpression. Scale bar: 200 μm. (C) Spheroid bodies are derived from Eca-109 CON, shN1, and ON cells. (D) Matrigel-coated Transwell assays were performed to determine the capability of cell invasion. Cells were stained, counted, and photographed (100 × ). (E) The expression of NANOG in Eca-109 CON, shN1, and ON cells was detected using protein blot analysis. (F) Comparison of rates of spheroid body formation of Eca-109 ON, CON, and shN1 groups (left). The number of Eca-109 ON, CON, and shN1 cells passing through the matrix (right). All the results were normalized to the blank, based on 3 repetitions (*P < .05; **P < .01; ***P < .001; ****P < .0001).

Abbreviations: ESCC, esophageal squamous cell carcinoma; BCA, Bicinchoninic acid; ON cells, NANOG overexpression cells.