Figure 5.
Esophageal squamous cell carcinoma (ESCC) cells with NEDD9 overexpression recruits G-MDSCs through CXCL8. (A) Correlation between myeloid-derived suppressor cells (MDSCs) and NEDD9 expression in patients with ESCC. (B) Correlation between G-MDSCs and CXCL8 expression in patients with ESCC. (C) A total of 2 × 105 tumor-infiltrating G-MDSCs from patients with ESCC were seeded into the upper chamber of Transwell inserts, and the numbers of cells that migrated into the lower chamber filled with supernatant from KYSE70-Vector and KYSE70-NEDD9shRNA cells were determined by flow cytometry. (D) Numbers of cells that migrated into the lower chamber filled with supernatant from KYSE450-Vector (KYSE450-pBpLV) and KYSE450-NEDD9 cells were determined by flow cytometry. The lower chamber was filled with supernatant from KYSE450-NEDD9 in the presence of anti-CXCL8-neutralizing antibody or an isotype-matched IgG. (E) Numbers of G-MDSCs that migrated into the lower chamber filled with different concentrations of CXCL8 were counted. (F–G) Kinetics of tumor growth in vivo (5 mice in each group). Tumor volume was measured every second day and tumor weight was evaluated. (H) Accumulation of transferred G-MDSCs in tumor-bearing mice treated with Reparixin, measured by flow cytometry. *P < 0.05; **P < 0.01; ***P < 0.001.