Figure 3.

FAM3C is involved in tumor cell EMT induced by Edu-Neus. (A) FAM3C treatment decreased E-cad expression and increased Vim expression in MKN28 and MKN45 cells in a dose-dependent manner. (B) RNA-seq analysis showed FAM3C mRNA was increased by more than 140 times in Edu-Neus. (C, D) Co-culture with MKN28 or MKN45 cells increased FAM3C level (C) or production (D) in neutrophils. (E, F) Blockage of FAM3C with AB72182 reversed the enhanced-invasiveness (E, ×100) or induced-EMT (F) of tumor cells mediated by Edu-Neus. (G) FAM3C immunostaining was detected in TANs in human gastric tumor tissues and cancer emboli with IHC (green arrow), but it was not detected in neutrophils in normal stomach tissues (blank arrow). (H) FAM3C mRNA levels in gastric tumor tissues (T) were much higher than that in corresponding normal stomach tissues (N). (I) FAM3C level in gastric tumors was related to TNM staging of gastric cancer although it was not significant (P = 0.063). (J) FAM3C level in tumors was negatively associated with cumulative survival of gastric cancer patients (P = 0.022). (K) FAM3C treatment up-regulated p-JNK as well as ZEB1 and Snail expression in MKN45 cells in a dose-dependent manner but exerted no marked effects on expression of p-ERK, p-Akt, Slug, and β-Catenin. (L) The expressions of E-cad, Vim and ZEB1 or Snail were reversed with JNK inhibitor treatment. (***, P < 0.001) (Neu, neutrophil; Edu-Neu, tumor-educated neutrophil).