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  • Title: [The mechanism of bystander effect in herpes simplex virus thymidine kinase/ganciclovir-mediated gene therapy].
    Author: Zhang L, Liu T, Cui Q, Gao J.
    Journal: Zhonghua Bing Li Xue Za Zhi; 1999 Dec; 28(6):440-4. PubMed ID: 11869559.
    Abstract:
    OBJECTIVE: To study the mechanism of bystander effect in herpes simplex virus thymidine kinase (HSV-TK)/Ganciclovir (GCV)-mediated gene therapy. METHODS: Recombinant retroviral vectors expressing HSV-TK and beta-Galactosidase (Lac Z) genes were constructed and transferred into pancreatic carcinoma cell line respectively. Cell counting was used to detect the growth inhibition rate of HSV-TK-transduced cells in presence of GCV. Taking Lac Z-transduced cells as bystander cells, the bystander effect was detected by MTT method, and its mechanism was studied by the experiments of supernatant shifting, Verapamil inhibition and ultrastructural observation. RESULTS: The growth inhibition rate of the HSV-TK-transduced cells in the presence of GCV was 92.1%, which was obviously 4.9% and 3.2% higher than of the non-and control vector-transduced cells. Mixed cells containing only 10% of HSV-TK-transduced cells showed 39.0% reduction of the proliferation, which meant there was an obvious bystander effect in the system. However this effect disappeared when transferring GCV-containing supernatant of HSV-TK-transduced cells to the parent cells and could be reduced significantly when verapamil was added in the medium, indicating that this bystander effect requires cell-cell contact. Gap junctions were observed existing between PC-2 cells by electron microscopy. CONCLUSION: The bystander effect in HSV-TK/GCV-mediated gene therapy occurs by transfer of GCV metabolite from cell to cell through gap junction.
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