Effect of 2-chloroadenosine on CXCL16-mediated inhibition of liver metastasis by SL4-CXCL16 cells. (A) M1 macrophage markers and TNF-α were detected by RT-PCR. GAPDH was used as the normalization control. N, normal; T, tumor. (B) Cytotoxicity of macrophage-derived TNF-α and recovery by TNF-α neutralizing antibody in SL4 cells. Cells (5 × 104 cells) were seeded in 24-well plates and a TNF-α neutralizing antibody added (2.5 μg/ml). RAW 264.7 cells (5 × 104 cells) were seeded in migration chambers and co-cultured. Cells were removed from the chambers and their viability was measured by WST-8 assay. *P <0.05, compared with control. #
P <0.05, compared with RAW co-culture. (C and D) Restoration of liver metastasis by CXCL16 expression in a macrophage depletion model. 2-Chloroadenosine was dissolved in saline and injected intraperitoneally (50 μg/100 μl) 24 h before tumor inoculation. *P <0.05. All experiments were repeated at least three times.