Effect of GCV on Neuroblastoma Cell Line Expressed by HSV-TK Gene with Retroviral Vector

ABSTRACT

Background: Gene transfer with vectors derived from murine retroviruses is restricted to cells which are proliferating and synthesizing DNA at the time of infection. Accordingly, selective introduction of genes encoding for susceptibility to otherwise nontoxic drugs (suicide genes) into proliferating tumor may be used to treat cancer. We investigated the efficacy of in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase (HSV-tk) gene followed by administration of the antiviral drug ganciclovir. METHODS: The LNC/tK vector was transfered in vitro into mouse Neuro 2a cell lines (ATCC) and the transduced cell lines were selected in G-418, 500microgram/ml, for 14 days. Onex104 cells were cultured in 96 well culture plates in increasing concentrations of ganciclovir for 72 hours. The sesitivity to ganciclivir of these HSV-tk transduced, G-418 selected cells was measured with MTT assay RESULTS: The survival of HSV-tk transduced 1x104 neuro 2a cell lines is 103+/-3.5%, 68+/-4.2%, 54+/-3.8%, 17+/-2.6%, 13+/-3.1% at the concentration of 0, 0.1, 1.0, 10, 20microgram/ml ganciclovir, respectively. And the survival of HSV-tk not transduced 1x104 neuro 2a cell lines is 100+/-4.5%, 97+/-5.6%, 104+/-3.5%, 106+/-3.8%, 101+/-4.2%. CONCLUSION: We concluded that in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase gene followed by administration of the antiviral drug ganciclovir is very effective.