title: Overexpression of coxsackie and adenovirus receptor inhibit growth of human bladder cancer cell in vitro and in vivo.
abstract: OBJECTIVE
  To study the effect of the overexpression of coxsackie and the adenovirus receptor (CAR) on the growth of the human bladder cancer cell in vitro and in vivo.
  
  
  METHODS
  A retroviral vector pLXSN-CAR expressing CAR was constructed and confirmed by restriction enzyme mapping. The pLXSN-CAR vector and control vector pLXSN were transfected into the PT67 packaging cell line to generate retrovirus with high titer. The CAR-negative T24 cell was infected with the pLXSN-CAR and the pLXSN retrovirus, respectively. The positive clone cells were selected with G418 for 2 weeks. The expression level of the CAR protein was detected by Western blot assay. T24 cell growth in vitro was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Anchorage-independent growth was measured by soft-agar colony formation assay. In vivo cell growth was determined by a nude mice xenograft model.
  
  
  RESULTS
  The pLXSN-CAR vector containing full-length CAR cDNA was successfully constructed. Western blot analysis showed that a 46 kDa specific band was found in pLXSN-CA-transfected T24 cells. MTT assay identified the growth inhibition of T24/pLXSN-CAR cells. The cell colony forming ability of T24/pLXSN-CAR cells was significantly lower than that of T24/pLXSN and parental T24 cells. There was a reduction in the tumor size in the T24/pLXSN-CAR group as compared with that of the T24/pLXSN group and parental T24 group.
  
  
  CONCLUSIONS
  The overexpression of CAR in T24 bladder cancer cells can inhibit cell growth both in vitro and in vivo.
authors.affiliation: Institute of Urology, First Hospital of Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Lin-Lin
authors.initials: LL
authors.lastname: Zhang
authors.forename: Da-Lin
authors.initials: DL
authors.lastname: He
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Guo-Dong
authors.initials: GD
authors.lastname: Zhu
authors.forename: Dong
authors.initials: D
authors.lastname: Zhang
authors.forename: Xin-Yang
authors.initials: XY
authors.lastname: Wang
mesh_terms: D000818:Animals; D002460:Cell Line; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology
chemical_list: C568771:CLMP protein, human; C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011991:Receptors, Virus