摘要
AIM： To construct the retroviral vector of p^125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapy in vitro. METHODS： A hammerhead ribozyme DNA targeting p^125FAK mRNA from nt 1010 to nt 1032 was synthesized and recombinated into the retroviral vector pLXSN forming pLRZXSN recon. Using the lipofectin-mediated DNA transfection technique, pLRZXSN was introduced into BGC-823 cells. The effects of ribozyme on the growth of BGC-823 cells and apoptosis were studied by cell colony assay, flow cytometry （FCM）, reverse transcriptasepolymerase chain reaction （RT-PCR）, detection of DNA fragmentation and electron microscopy. RESULTS： The number of BGC-823 cell colonies was inhibited by 56% after the cells were treated for 48 h. The cell proliferation was inhibited effectively by p^125FAK ribozyme and the inhibitory effect depended on the concentration and the time of incubation. The expression of p^125FAK mRNA and protein p^125 decreased sharply in BGC-823 cells treated with p^125FAK ribozyme. The characteristics of apoptosis, namely sub-G1 peak, DNA fragmentation and morphological changes, were revealed in BGC-823 cells treated with p^125FAK ribozyme. CONCLUSION： p^125FAK ribozyme decreases p^125FAK gene expression and induces apoptosis of human gastric cancer cells in vitro.
瞄准：构造制动火箭 p (125FAK ) 的病毒的向量特定的 ribozyme 基因并且在 BGC-823 探索 ribozyme 的可行性基因治疗在试管内。方法：从 nt 的 A 撞木鲛 ribozyme DNA 指向 p (125FAK ) mRNA 1010 到 nt， 1032 被综合并且重新结合进制动火箭病毒的向量 pLXSN 形成 pLRZXSN 交换子。用调停 lipofectin 的 DNA transfection 技术， pLRZXSN 被介绍进 BGC-823 房间。BGC-823 细胞和 apoptosis 的生长上的 ribozyme 的效果被细胞殖民地试金，流动血细胞计数(FCM ) ，反向的 transcriptase 聚合酶链反应(RT-PCR ) ， DNA 破碎的察觉和电子显微镜学学习。结果：在房间为 48 h 被对待以后， BGC-823 房间殖民地的数字被 56% 禁止。房间增长被 ribozyme 和禁止的效果取决于的 p (125FAK ) 有效地禁止集中和孵化的时间。p (125FAK ) 的表示 mRNA 和蛋白质 P (125FAK ) 在与 p (125FAK ) 对待的 BGC-823 房间严厉地减少了 ribozyme。apoptosis 的特征，也就是 sub-G1 山峰， DNA 破碎和词法变化，在与 p (125FAK ) 对待的 BGC-823 房间被揭示 ribozyme。结论：p (125FAK ) ribozyme 减少 p (125FAK ) 基因表示并且导致人的胃的癌症房间的 apoptosis 在试管内。
基金
Supported by the Natural Science Foundation of Fujian Province,No.C0010015
