为探讨腺病毒载体介导的外源性Rb基因导入对膀胱癌细胞生长的抑制作用,为膀胱癌的基因治疗提供实验依据,构建了Rb基因的复制缺陷型重组腺病毒载体.体外转染人膀胱癌细胞株EJ.应用免疫组化法、免疫印迹及聚合酶链反应技术检测外源性Rb基因腺病毒载体的转染效率及Rb基因表达效果;用流式细胞仪分析细胞周期;以细胞计数及同位素掺入技术观察外源性Rb基因对EJ细胞生长的抑制效果.结果显示腺病毒载体可有效地将外源基因导入EJ细胞.Rb基因重组腺病毒载体转染细胞后,胞内DNA合成减少,细胞生长受到抑制.流式细胞仪分析显示68%的EJ细胞生长停滞在GO/GI期.结果提示,外源性Rb基因重组腺病毒载体转染膀胱癌细胞可有效抑制细胞生长

In order to study the bladder carcinoma cell growth suppression by introduction of foreign retinoblastoma (Rb) gene and explore a gene therapy approach for bladder cancer, a replication-deficient adenovirus vector encoding a wild-type Rb, AdCMVRb, was constructed and transfected into the cultured human bladder carcinoma cell line EJ. The efficiency of gene transfection and expression was detected by immunochemical staining, Western blotting and polymerase chain reaction. The role of Rb in suppressing EJ growth was observed by cell-counting, [3H]thymidine incorporation and flow cytometry. The results showed that wild-type Rb gene could be transfected effectively into cultured EJ with Ad-CMVRb and could arrest the cells at GO/Gl phases of the cell cycle, leading to inhibition of DNA synthesis. The results demonstrated the potential of adenovirus-mediated Rb gene therapy for bladder cancer.

R737.140.5 R394

国家自然科学基金(39640005)资肋;广东省科教兴医“五个一”工程重点课题,粤卫科[1996]20号