title: The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis.
abstract: OBJECTIVE
  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 23.5 ± 5.1 μmol·L(-1) for BGC-823 cells and 35.6 ± 7.6 μmol·L(-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.
  
  
  CONCLUSIONS
  DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Sen-Sen
authors.initials: SS
authors.lastname: Lin
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Fan
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China. Electronic address: cpusunli@126.com.
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Li
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Ren-Ping
authors.initials: RP
authors.lastname: Zhao
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China.
authors.forename: Lu-Yong
authors.initials: LY
authors.lastname: Zhang
authors.affiliation: Department of Complex Prescription of TCM, China Pharmaceutical University, Nanjing 211198, China.
authors.forename: Bo-Yang
authors.initials: BY
authors.lastname: Yu
authors.affiliation: New Drug Screen Center, China Pharmaceutical University, Nanjing 210009, China. Electronic address: yuanst@aliyun.com.
authors.forename: Sheng-Tao
authors.initials: ST
authors.lastname: Yuan
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects*; D018946:Chemokine CCL5 / Q000032:analysis*; D015536:Down-Regulation; D006801:Humans; D009362:Neoplasm Metastasis / Q000188:drug therapy; D019713:Receptors, CCR5 / Q000032:analysis*; D012503:Saponins / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000473:pathology*; D014407:Tumor Cells, Cultured
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C584933:CCR5 protein, human; D018946:Chemokine CCL5; D019713:Receptors, CCR5; D012503:Saponins; C578958:ruscogenin-1-O-(glucopyranosyl-(1-2))(xylopyranosyl-(1-3))fucopyranoside
keywords: BGC-823; CCL5; CCR5; CXCR4; Gastric cancer cell migration; HGC-27; Liriope muscari; Saponin DT-13