The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis

AIM:To investigate the effect of DT-13 on gastric cancer cell migration,and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13.METHODS:Growth inhibition of DT-13 was analyzed by the MTT assay.Cell migration was measured by the scratch-wound assay and transwell double chamber assay.To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13,chemokine receptors that are involved in cancer metastasis(CCR2,CCR5,CCR7,CXCR4,and CXCR6) were detected by conventional PCR.The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot,respectively.The secretion of CCL5(ligand of CCR5) and SDF-1(ligand of CXCR4) were detected by enzyme-linked immunosorbent assay(ELISA).RESULTS:DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner,and the estimated IC50 value for 24 h treatment was 23.55.1 molL-1 for BGC-823 cells and 35.67.6 molL-1 for HGC-27 cells.DT-13 also significantly decreased gastric cancer cell migration.DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells,and moderately reduced the expression of CXCR4.Similar to the results of gene expression,significant down-regulation of CCR5 protein was observed,but CXCR4 protein levels were much less affected.CCL5 secretion,but not SDF-1 production,was inhibited by DT-13.CONCLUSION:DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.