论文部分内容阅读
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 investigate the inhibitory effect of adenovirus vector-mediated exogenous Rb gene transfection on the growth of human bladder cancer cells and to provide an experimental basis for the gene therapy of bladder cancer, we constructed the replication-deficient recombinant adenovirus vector of Rb gene.In vitro transfection Bladder cancer cell strain EJ. The transfection efficiency and Rb gene expression of exogenous Rb gene adenovirus vector were detected by immunohistochemistry, Western blot and polymerase chain reaction (PCR); cell cycle was analyzed by flow cytometry; Counting and isotope incorporation technology to observe the inhibitory effect of exogenous Rb gene on EJ cell growth.The results showed that adenovirus vector can effectively introduce foreign gene into EJ cells.Rb gene recombinant adenovirus vector transfected cells, Flow cytometry analysis showed that 68% of EJ cells arrested in GO / GI phase.The results suggest that transfection of exogenous Rb gene recombinant adenovirus vector into bladder cancer cells can effectively inhibit cell growth