title: CCR4 mediates CCL17 (TARC)-induced migration of human colon cancer cells via RhoA/Rho-kinase signaling.
abstract: BACKGROUND
  Accumulating data suggest a role of chemokines in tumor cell metastasis. CCR4 has been implicated in hematologic malignancies and recently also in solid tumors. Herein, we hypothesized that CCR4 might be expressed and support migration of colon cancer cells.
  
  
  METHODS
  We used quantitative RT-PCR and flow cytometry to determine mRNA and surface expression of CCR4 on colon cancer cell lines (HT-29) and (AZ-97). Total RhoA and active RhoA protein levels in CCL17-stimulated colon cancer cells were quantified using ELISA and G-LISA assays. Migration assays were performed to evaluate colon cancer cells chemotaxis. In vitro tumor growth was assessed using proliferation assay.
  
  
  RESULTS
  Our results show clear-cut mRNA levels and surface expression of CCR4 on a colon cancer cell line (HT-29) and on tumor cells (AZ-97). CCR4 ligand CCL17 (TARC) was a potent stimulator of colon cancer cell migration. This CCL17-induced colon cancer cell migration was inhibited by pre-incubation of the colon cancer cells with an antibody directed against CCR4 or an antagonist against CCR4. CCL17-induced signaling in colon cancer cells revealed that CCL17 increased mRNA formation of RhoA-C in colon cancer cells. Our results also found that CCL17 increased total RhoA and active RhoA protein levels in colon cancer cells. The Rho-kinase inhibitor Y-27632 abolished CCL17-induced colon cancer cell chemotaxis. In addition, inhibition of isoprenylation by GGTI-2133 markedly reduced colon cancer cell migration triggered by CCL17.
  
  
  CONCLUSIONS
  Our novel data indicate for the first time that the CCL17-CCR4 axis might be involved in the spread of colon cancer cells.
authors: Department of Clinical Sciences, Malmö, Section of Surgery, Skåne University Hospital, Lund University, 20502, Malmö, Sweden.
authors: Amr A
authors: AA
authors: Al-haidari
authors: Ingvar
authors: I
authors: Syk
authors: Karin
authors: K
authors: Jirström
authors: Henrik
authors: H
authors: Thorlacius
mesh_terms: D000368:Aged; D000577:Amides / Q000494:pharmacology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D054414:Chemokine CCL17 / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D054398:Receptors, CCR4 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000577:Amides; C106230:CCL17 protein, human; C515354:CCR4 protein, human; D054414:Chemokine CCL17; C547856:GGTI-2133; D007093:Imidazoles; D009281:Naphthalenes; D011725:Pyridines; D012333:RNA, Messenger; D054398:Receptors, CCR4; C066737:RHOA protein, human; C108830:Y 27632; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D007930:Leucine
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