title: A Lentiviral CXCR4 overexpression and knockdown model in colorectal cancer cell lines reveals plerixafor-dependent suppression of SDF-1α-induced migration and invasion.
abstract: BACKGROUND
  The development of distant metastasis is associated with poor outcome in patients with colorectal cancer (CRC). The stromal cell-derived factor-1 (SDF-1) and its receptor CXC chemokine receptor 4 (CXCR4) have pivotal roles in the chemotaxis of migrating tumor cells during metastasis. Thus, hampering the SDF-1/CXCR4 cross-talk is a promising strategy to suppress metastasis.
  
  
  METHODS
  We investigated the invasive behavior of the lentivirally CXCR4 overexpressing CRC cell lines SW480, SW620 and RKO in chemotaxis and invasion assays toward an SDF-1α gradient. Low endogenous CXCR4 expression levels were determined by quantitative realtime polymerase chain reaction (PCR) and fluorescence-activated cell sorting (FACS) analyses.
  
  
  RESULTS
  A lentiviral CXCR4 overexpression and knockdown model was established in these CRC cells. In transwell migration assays, CXCR4 overexpression favored chemotaxis and invasion of cells in all 3 lines depending on an SDF-1α gradient (p < 0.001 vs. untransduced cells). Functional CXCR4 knockdown using lentiviral short hairpin RNA (shRNA) vectors significantly decreased the migration behavior in CRC cell lines (p < 0.001), confirming a CXCR4-specific effect. Pharmacologic inhibition of the SDF-1α/CXCR4 interaction by the bicyclam Plerixafor(TM) at 100 μM significantly abrogated CXCR4-dependent migration and invasion through Matrigel(TM) (SW480, SW620, RKO; p < 0.05).
  
  
  CONCLUSIONS
  Our results indicate that a CXCR4-antagonistic therapy might prevent tumor cell dissemination and metastasis in CRC patients, consequently improving survival.
authors: Molecular Oncology of Solid Tumors, DKFZ (German Cancer Research Center), Heidelberg, Germany.
authors: Doreen
authors: D
authors: Heckmann
authors: Stephanie
authors: S
authors: Laufs
authors: Patrick
authors: P
authors: Maier
authors: Manuela
authors: M
authors: Zucknick
authors: Frank A
authors: FA
authors: Giordano
authors: Marlon R
authors: MR
authors: Veldwijk
authors: Volker
authors: V
authors: Eckstein
authors: Frederik
authors: F
authors: Wenz
authors: W Jens
authors: WJ
authors: Zeller
authors: Stefan
authors: S
authors: Fruehauf
authors: Heike
authors: H
authors: Allgayer
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D001596:Benzylamines; D045744:Cell Line, Tumor; D054377:Chemokine CXCL12 / Q000235:genetics*; D002633:Chemotaxis / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D000080027:Cyclams; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics*; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006571:Heterocyclic Compounds / Q000494:pharmacology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009361:Neoplasm Invasiveness / Q000235:genetics* / Q000473:pathology; D020239:Receptor Cross-Talk / Q000187:drug effects; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: D019380:Anti-HIV Agents; D001596:Benzylamines; C515255:CXCL12 protein, human; C527276:CXCR4 protein, human; D054377:Chemokine CXCL12; D000080027:Cyclams; D006571:Heterocyclic Compounds; D019718:Receptors, CXCR4; C088327:plerixafor