Construction of Urothelium-Specific Recombinant Adenovirus and Its Inhibition in Bladder Cancer Cell
Construction of Urothelium-Specific Recombinant Adenovirus and Its Inhibition in Bladder Cancer Cell
复制标题
尿路上皮特异性重组腺病毒的构建及其对膀胱癌细胞的抑制作用
DOI：
10.1159/000200802
复制
发表时间：
2009-01-01
影响因子：
1.6
通讯作者：
Rodriguez, Ronald
中科院分区：
文献类型：
--
作者：
He, Xiang-Dong;Wang, Zhi-Ping;Rodriguez, Ronald
目的：构建尿路上皮特异性重组腺病毒并研究其对膀胱癌细胞的抑制作用。方法：采用RT-PCR方法检测hUPII和柯萨奇腺病毒受体在多个细胞系上的表达规律。采用瞬时转染和荧光素酶检测法检测hUPII启动子的组织特异性。构建了重组腺病毒Ad-UPII-E1A和Ad-UPII-Null。限制性酶切实验和PCR证实了正确的结构。重组腺病毒感染BIU-87细胞后，用Western blot检测其表达的腺病毒E1A蛋白。研究了重组腺病毒Ad-UPII-E1A对膀胱癌细胞株BIU-87的抑制作用。结果：HUPII和CAR在膀胱癌细胞株BIU-87中均有表达，且HUPII启动子具有高活性。利用细菌同源重组技术，将hUPII启动子和E1A基因插入重组5型腺病毒基因组中。重组腺病毒Ad-UPII-E1A感染的BIU-87细胞样品中E1A蛋白明显阳性。MTT实验证实重组腺病毒Ad-UPII-E1A抑制膀胱癌细胞BIU-87的生长。结论：hUPII启动子具有较高的组织特异性。构建并确认重组腺病毒Ad-UPII-E1A和Ad-UPII-Null。重组腺病毒Ad-UPII-E1A对膀胱癌细胞株BIU-87有抑制作用。
Aim: To construct urothelium-specific recombinant adenovirus and investigate its inhibition in bladder cancer cell. Methods: RT-PCR analysis was used to determine expression patterns of hUPII and coxsackie adenovirus receptor on multiple cell lines. Transient transfection and luciferase detecting assay were used to detect tissue specificity of the hUPII promoter. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed. Restrictive enzyme digestion assay and PCR confirmed the correct construction. The adenovirus E1A protein expressed in BIU-87 was tested by Western blot after cells were infected with recombinant adenovirus. Recombinant adenovirus Ad-UPII-E1A was tested for its inhibition in bladder cancer cell line BIU-87. Results: HUPII and CAR were expressed and the hUPII promoter is highly active in bladder cancer cell line BIU-87. Using homologous recombination in bacteria technology, the hUPII promoter and E1A gene were inserted into the genome of type 5 recombinant adenovirus. The E1A protein was markedly positive in the samples of BIU-87 cells infected with recombinant adenovirus Ad-UPII-E1A. MTT assay demonstrated recombinant adenovirus Ad-UPII-E1A inhibited bladder cancer cell BIU-87 growth. Conclusion: The hUPII promoter shows high tissue specificity. Recombinant adenovirus Ad-UPII-E1A and Ad-UPII-Null were constructed and confirmed. Recombinant adenovirus Ad-UPII-E1A is effective in inhibition in bladder cancer cell line BIU-87.