ANXA10 induction by interaction with tumor-associated macrophages promotes the growth of esophageal squamous cell carcinoma

Tumor-associated macrophages (TAMs) have important roles in the progression, angiogenesis and motility of various cancers. To study the effects of TAMs on the tumor microenvironment of ESCCs, we constructed an in vitro system for the differentiation of peripheral blood monocyte (PBMo)-derived macrophages into TAM-like PBMo-derived macrophages. We established a co-culture assay using a human ESCC cell line and TAM-like PBMo-derived macrophages and we performed a cDNA microarray analysis between monocultured and co-cultured ESCC cell lines. Our qRT-PCR confirmed that in the co-cultured ESCC cell lines, CYP1A1, DHRS3, ANXA10, KLK6 and CYP1B1 mRNA were highly up-regulated; AMTN and IGFL1 mRNA were down-regulated. We observed that a high ANXA10 expression was closely associated with the depth of invasion and high numbers of infiltrating CD68+ and CD204+ TAMs. ESCC patients with high ANXA10 expression were significantly correlated with poor disease-free survival (p = 0.0216). ANXA10 knockdown using siRNA significantly suppressed the cell growth of TE-8 and TE-9 ESCC cells by inhibiting Akt and Erk1/2 signaling pathways. We confirmed that ANXA10 overexpression induced the cell growth of TE-15 ESCC cells by activating Akt and Erk1/2 phosphorylation. These results suggest that ANXA10 induced by TAMs in the tumor microenvironment is associated with aberrant growth and a poor prognosis in human ESCC tissues. PBMos were cultured with macrophage-colony stimulating factor (M-CSF) for 6 days to induce macrophages. We added 50% condition media of human ESCC cell line (TE-8 and TE-9) to culture for 2 days to induce TAM-like polarization. We established co-culture assay using human ESCC cell line (TE-8 and TE-9) and TAM-like macrophages and performed cDNA microarray analysis between monocultured and co-cultured ESCC cell lines.