GASC1 Promotes Stemness of Esophageal Squamous Cell Carcinoma via NOTCH1 Promoter Demethylation

Figure 7

NOTCH1 is required for GASC1-induced CSC-like properties in ESCC. Sphere forming ability of shNOTCH1 and shRNA scramble KYSE150 cells was analyzed. (a) One representative micrograph is shown. Scale bar represents 20 μm. (b) Sphere forming efficiency is shown as a histogram. (c) shGASC1 KYSE150 cells before and after NOTCH1 vector infection were capable of self-renewal in vitro, as shown by similar esosphere-initiating capacity in serial passages. (d) The percentage of ALDH⁺ cells in shGASC1 KYAE150 cells before and after infection of NOTCH1 vector was analyzed by flow cytometry. Sphere forming ability (e) and ALDH⁺ cell frequency (f) of KYSE150 cells (with or without NOTCH1 overexpression) before and after treatment with CA (10 μM) were analyzed. (g) Tumor volumes were measured after implantation of purified ALDH⁺ KYSE150 cells (with or without NOTCH1 overexpression) treated with CA (10 mg/kg). Data are represented as means ± SD. = P < 0.05.

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