title: Chemotherapeutic agents attenuate CXCL12-mediated migration of colon cancer cells by selecting for CXCR4-negative cells and increasing peptidase CD26.
abstract: BACKGROUND
  Recurrence of colorectal cancer (CRC) may arise due to the persistence of drug-resistant and cancer-initiating cells that survive exposure to chemotherapy. Proteins responsible for this recurrence include the chemokine receptor CXCR4, which is known to enable CRC metastasis, as well as the cancer-initiating cell marker and peptidase CD26, which terminates activity of its chemokine CXCL12.
  
  
  METHODS
  We evaluated the expression and function of CXCR4 and CD26 in colon cancer cell lines and xenografts following treatment with common chemotherapies using radioligand binding, flow cytometry, immunofluorescence, and enzymatic assays.
  
  
  RESULTS
  5-Fluorouracil, oxaliplatin and SN-38 (the active metabolite of irinotecan), as well as cisplatin, methotrexate and vinblastine, each caused decreases in cell-surface CXCR4 and concomitant increases in CD26 on HT-29, T84, HRT-18, SW480 and SW620 CRC cell lines. Flow cytometry indicated that the decline in CXCR4 was associated with a significant loss of CXCR4+/CD26- cells. Elevations in CD26 were paralleled by increases in both the intrinsic dipeptidyl peptidase activity of CD26 as well as its capacity to bind extracellular adenosine deaminase. Orthotopic HT-29 xenografts treated with standard CRC chemotherapeutics 5-fluorouracil, irinotecan, or oxaliplatin showed dramatic increases in CD26 compared to untreated tumors. Consistent with the loss of CXCR4 and gain in CD26, migratory responses to exogenous CXCL12 were eliminated in cells pretreated with cytotoxic agents, although cells retained basal motility. Analysis of cancer-initiating cell CD44 and CD133 subsets revealed drug-dependent responses of CD26/CD44/CD133 populations, suggesting that the benefits of combining standard chemotherapies 5-fluoruracil and oxaliplatin may be derived from their complementary elimination of cell populations.
  
  
  CONCLUSIONS
  Our results indicate that conventional anticancer agents may act to inhibit chemokine-mediated migration through eradication of CXCR4+ cells and attenuation of chemokine gradients through elevation of CD26 activity.
authors.affiliation: School of Pharmacy, University of Waterloo, 200 University Ave. W., Waterloo, ON, N2L 3G1, Canada. murray.cutler.jr@gmail.com.|Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. murray.cutler.jr@gmail.com.
authors.forename: Murray J
authors.initials: MJ
authors.lastname: Cutler
authors.affiliation: Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. lowthers@simip.com.|Sim & McBurney/Sim Lowman Ashton & McKay LLP, Toronto, Ontario, Canada. lowthers@simip.com.
authors.forename: Erica L
authors.initials: EL
authors.lastname: Lowthers
authors.affiliation: School of Pharmacy, University of Waterloo, 200 University Ave. W., Waterloo, ON, N2L 3G1, Canada. c25richard@uwaterloo.ca.|Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. c25richard@uwaterloo.ca.
authors.forename: Cynthia L
authors.initials: CL
authors.lastname: Richard
authors.affiliation: Department of Statistics and Actuarial Science, University of Waterloo, Waterloo, Ontario, Canada. hajducek@gmail.com.
authors.forename: Dagmar M
authors.initials: DM
authors.lastname: Hajducek
authors.affiliation: School of Pharmacy, University of Waterloo, 200 University Ave. W., Waterloo, ON, N2L 3G1, Canada. paul.spagnuolo@uwaterloo.ca.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Spagnuolo
authors.affiliation: School of Pharmacy, University of Waterloo, 200 University Ave. W., Waterloo, ON, N2L 3G1, Canada. jonathan.blay@uwaterloo.ca.|Department of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. jonathan.blay@uwaterloo.ca.|Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada. jonathan.blay@uwaterloo.ca.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Blay
mesh_terms: D000818:Animals; D002166:Camptothecin / Q000008:administration & dosage / Q000031:analogs & derivatives; D063646:Carcinogenesis / Q000187:drug effects; D019070:Cell Lineage; D002465:Cell Movement / Q000187:drug effects; D054377:Chemokine CXCL12 / Q000096:biosynthesis* / Q000235:genetics; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D018819:Dipeptidyl Peptidase 4 / Q000096:biosynthesis* / Q000235:genetics; D005472:Fluorouracil / Q000008:administration & dosage; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D019073:HT29 Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics; D000077146:Irinotecan; D051379:Mice; D009362:Neoplasm Metastasis; D019718:Receptors, CXCR4 / Q000096:biosynthesis* / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: C515255:CXCL12 protein, human; C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; D018960:Hyaluronan Receptors; D019718:Receptors, CXCR4; D000077146:Irinotecan; C042807:DPP4 protein, human; D018819:Dipeptidyl Peptidase 4; D005472:Fluorouracil; D002166:Camptothecin