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HSV-tk/GCV retrovirus-mediated effects on liver cancer cells


Of HCC9204 / {kfB] roar significant killing effect, 724 The GCV of the wild-type HCC9204fm no significant cell toxicity (P O.05) (Figure 4) Time (h) not the same asked GCV loop 3 killing effect on liver cancer cells, and numerous (dl around 4GCV tumor inhibition in vivo t discussion, we will HsV ~ tk gene cloned human pDOR-neo retroviral vector, the successfully constructed HSV-tk gene in eukaryotic expression vector pDORtk. by packaging cells packaged into HSV-tk gene with replication-defective virus, and infected human hepatoma cells Hcc9204, dot blotting results confirmed that: HSV-tk gene was integrated DNA in human HCC9204 cells. and there is mRNA stable expression. transduction of the HSV-tk gene HcC9204/tk cells, can be very low GCV concentrations of anti-(Ic50 to 1.24rag / L), and the killing effect of GCV in a dose-dependent and time-dependent characteristics : The GcV of wild-type HCC9204 cells without significant toxicity,[link widoczny dla zalogowanych], indicating that HSV-tk gene product to be effective in GCV into cytotoxic products, resulting in cell death genetically. HsV-tk/GcV gene therapy system in nude exvivo level (finger gene transferred into the target cells in vitro, then transferred to the target cells in vivo transgenic observation) also obtained similar results, indicating Gcv effective anti-HSV-tk gene transfer in vivo liver cancer solid tumor. HSV-tk gene is the most common susceptibility gene, one of the herpes-like virus and mammalian cells are present encoding thymidine kinase (tk) gene,[link widoczny dla zalogowanych], tk-specific mammalian cells is very strong,[link widoczny dla zalogowanych], very narrow substrate range, only catalytic deoxygenation chest glucoside phosphorylation, the HSV-tk addition, a number of nucleoside analogues can be catalyzed (such as GCV, ACV, BVdU, etc.) phosphorylation, and its product can inhibit cellular DNA synthesis,[link widoczny dla zalogowanych], leading to cell death HsV-tk gene transfected hepatoma cells transfected hepatoma cells with and without mixed culture, and exposed to GCV, not only the transfected cells were killed and their co-culture of transfected cells can not be killed,[link widoczny dla zalogowanych], thus expanding the killing effect. This phenomenon known as the / tk and HcC9204 hybrid cells (at 1:5 mixture) to form tumors have some degree of inhibition. but HcC9204 ~ tumor was no significant difference, which may be mixed with the proportion of cells in a variety of complex mechanisms and in vivo, the specific needs further study because HSV-tk/GCV gene therapy system has many advantages: such as low dose of anti-gene can significantly GcV transfected cells; next to the killing effects: selective destruction of rapid proliferation of transfected cells. I believe that with gene transfer vector and the transfer of technology, continuous improvement, drug sensitivity gene therapy will be effective gene therapy methods.