Combined gene therapy with suicide gene and interleukin-12 is more efficient than therapy with one gene alone in a murine model of hepatocellular carcinoma
Section snippets
Animals
Female BALB/c mice aged 6–8 weeks were obtained from Charles Rivers Laboratories (Barcelona, Spain). During the experimental period animals were housed in standard conditions, and all animal procedures were performed according to approved protocols and in accordance with recommendations for proper care and use of laboratory animals.
Cell lines and cell culture
Murine BNL 1MEA.7R.1 (BNL) methylcholanthrene transformed liver cell line and 293 were obtained from American Type Culture Collection (Rockville, MD, USA). The cells
Gene transfer efficiency in BNL cells with Ad vectors
As shown in Fig. 1, BNL cells are relatively resistant to infection by AdCMVlacZ. At moi 100, only a few cells were infected, and an increase of moi to 1000 raised the number of transduced cells up to 10–15%. Intratumoral injection of AdCMVlacZ 5×109 resulted in transduction of cells surrounding the site of injection. More distant parts of the tumor were not transduced.
IL-12 production after infection of BNL cells with AdCMVIL-12
Supernatants collected from non-infected BNL cells and cells infected in vitro with AdCMVlacZ at moi 100 and 1000 did not show
Discussion
Immunogene therapy represents remarkable progress in the field of cancer gene therapy. Many cytokines have been studied so far, including IL-2, IL-4, IL-7, IL-10, IL-12, IFN-γ, TNF-α, and GM-CSF 6., 7., 8., 9.. Among these cytokines, IL-12 is of particular interest because of its critical role in the stimulation of cellmediated immunity (10). In addition, IL-12 has antiangiogenic effect by induction of chemokines such as IP-10 and Mig 14., 18.. IL-12 has been shown to be very effective in
Acknowledgements
This work was supported by SAF 98–0146 from CICYT. M.D. is supported by a fellowship from the Fundación Empresa Universidad de Navarra, R.B. by Beca del Programa de Formación de Investigadores del Departamento de Educacion, Universidades e Investigacion del Gobiermo Vasco, and G.M. by an Ines Bemberg Grant. We thank Pilar Alzuguren for her excellent technical assistance.
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2005, Molecular TherapyCitation Excerpt :To strengthen the idea of targeting gene-virotherapy, the concept of a targeting dual gene-virotherapy strategy was devised, which uses a combination of two different therapeutic genes in oncolytic adenoviral vectors. It has been suggested that combination of two therapeutic genes mediated by replication-defective vectors has exerted synergic or compensatory anti-tumor activity [22–28]. Therefore, it is interesting to study whether a synergic or compensatory effect could be induced by combination of different therapeutic genes in oncolytic vectors.
Combination electro-gene therapy using herpes virus thymidine kinase and interleukin-12 expression plasmids is highly efficient against murine carcinomas in vivo
2004, Molecular TherapyCitation Excerpt :Interestingly, Hall et al. demonstrated the increased expression of Fas and FasL in tumor cells as the mechanism of enhanced apoptosis in tumor site [57]. Similarly, Drozdozik showed that a combination therapy using an Av system is more effective for the treatment of established hepatocellular carcinoma (HCC) than therapy with a single vector [58]. However, the treatment was not repeated and complete regression in the subcutaneous model of HCC was observed in only very few cases.
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