Cyr61 promotes CD204 expression and migration of macrophages via MEK/Erk pathway in the microenvironment of human esophageal squamous cell carcinoma

The authors previously suggested that CD204 was a useful marker for tumor-associated macrophages (TAMs) of esophageal squamous cell carcinoma (ESCC). In this study, they discovered that within ESCC microenvironment not only cancer cells but also TAMs expressed cysteine-rich, angiogenic inducer, 61 (Cyr61), induced by conditioned media of ESCC cells. Levels of Cyr61 expression were associated with CD204+ macrophage infiltration in ESCC tissue. Cyr61 promoted CD204 expression and migration of macrophages via MEK/Erk pathway in vitro. To identify the molecules specifically upregulated in TAM-like THP-1 cells, we performed a cDNA microarray analysis using RNAs from THP-1 stimulated with 200 nM TPA or with 200 nM TPA followed by 50% TE-8CM. THP-1 human acute monocytic leukemia cell line was treated with 200 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) for 2 days to induce macrophage-like differentiation, then exposed to 50%Conditioned medium of TE-8 ESCC cell line (TE-8CM) for 2 days. Differentially expressed genes between MΦ-like and TAM-like THP-1 cells were analysed by cDNA microarray. TE-8CM was prepared by plating 5 × 106 tumor cells in 10 mL complete medium in 100-mm dishes for 24 hr, thereafter changing the medium to complete DMEM supplemented with 10% human AB serum instead of fetal bovine serum. After 2–3 days, the supernatants were harvested, centrifuged and stored in aliquots at −80°C.