【摘 要】
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AIM: To construct the retroviral vector of p125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapyin vitro.METHODS: A
【机 构】
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Department of Oncology, Affiliated Union Hospital, Fujian Medical University, Fuzhou 350001, Fujian
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AIM: To construct the retroviral vector of p125FAK specific ribozyme genes and to explore the feasibility of ribozyme in BGC-823 gene therapyin vitro.METHODS: A hammerhead ribozyme DNA targeting p125FAK mRNA from nt 1010 to nt 1032 was synthesized and recombinated into the retroviral vector pLXSN forming pLRZ XSN 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 p125FAK ribozyme and the inhibitory effect depended on the concentration and the time of incubation. The expression of p125FAK mRNA and protein p125 decreased sharply in BGC-823 cells treated with p125FAK ribozyme. The characteristics of apoptosis, namely sub-G1 peak, DNA fragmentation and morphological changes, were revealed in BGC-823 cells treated with p125FAK ribozyme.CONCLUSION: p125FAK ribozyme decreases p125FAK gene expression and induces apoptosis of human gastric cancer cells in vitro.
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