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Aim:To study the effect of the overexpression of coxsackie and the adenovirusreceptor (CAR) on the growth of the human bladder cancer cell in vitro and invivo.Methods:A retroviral vector pLXSN-CAR expressing CAR was constructedand confirmed by restriction enzyme mapping.The pLXSN-CAR vector and con-trol vector pLXSN were transfected into the PT67 packaging cell line to generateretrovirus with high titer.The CAR-negative T24 cell was infected with the pLXSN-CAR and the pLXSN retrovirus,respectively.The positive clone cells wereselected with G418 for 2 weeks.The expression level of the CAR protein wasdetected by Western blot assay.T24 cell growth in vitro was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Anchor-age-independent growth was measured by soft-agar colony formation assay.Invivo cell growth was determined by a nude mice xenograft model.Results:ThepLXSN-CAR vector containing full-length CAR cDNA was successfullyconstructed.Western blot analysis showed that a 46 kDa specific band wasfound in pLXSN-CA-transfected T24 cells.MTT assay identified the growthinhibition 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 com-pared with that of the T24/pLXSN group and parental T24 group.Conclusion:The overexpression of CAR in T24 bladder cancer cells can inhibit cell growthboth in vitro and in vivo.
Aim: 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 invivo. Methods: A retroviral vector pLXSN-CAR expressing CAR was constructed and confirmed by restriction enzyme mapping. The pLXSN-CAR vector and con-trol 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 segmented with G418 for 2 weeks.The expression level of the CAR protein wasdetected by Western blot assay. T24 cell growth in vitro was determined by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay . Anchor-age-independent growth was measured by soft-agar colony formation assay. Invivo cell growth was determined by a nude mice xenograft model. Results: ThepLXSN-CAR vector containing full-length CAR cDNA was successfully constructed. Western blot an alysis showed that 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 com-pared with that of the T24 / pLXSN group and parental T24 group. Conflusion: The overexpression of CAR in T24 bladder cancer cells can inhibit cell growthboth in vitro and in vivo.