Improved retroviral suicide gene transfer in colon cancer cell lines after cell synchronization with methotrexate

• Main Authors: Nordlinger Bernard, Penna Christophe, Goere Diane, Noullet Severine, Capron Claude, Kraemer Aurore, Finzi Laetitia, Legagneux Josette, Emile Jean-Fançois, Malafosse Robert
• Format: Article
• Language: English
• Published: BMC 2011-10-01
• Series: Journal of Experimental & Clinical Cancer Research
• Online Access: http://www.jeccr.com/content/30/1/92

<p>Abstract</p> <p>Background</p> <p>Cancer gene therapy by retroviral vectors is mainly limited by the level of transduction. Retroviral gene transfer requires target cell division. Cell synchronization, obtained by drugs inducing a reversible inhibition of DNA synthesis, could therefore be proposed to precondition target cells to retroviral gene transfer. We tested whether drug-mediated cell synchronization could enhance the transfer efficiency of a retroviral-mediated gene encoding herpes simplex virus thymidine kinase (HSV-<it>tk</it>) in two colon cancer cell lines, DHDK12 and HT29.</p> <p>Methods</p> <p>Synchronization was induced by methotrexate (MTX), aracytin (ara-C) or aphidicolin. Gene transfer efficiency was assessed by the level of HSV-TK expression. Transduced cells were driven by ganciclovir (GCV) towards apoptosis that was assessed using annexin V labeling by quantitative flow cytometry.</p> <p>Results</p> <p>DHDK12 and HT29 cells were synchronized in S phase with MTX but not ara-C or aphidicolin. In synchronized DHDK12 and HT29 cells, the HSV-TK transduction rates were 2 and 1.5-fold higher than those obtained in control cells, respectively. Furthermore, the rate of apoptosis was increased two-fold in MTX-treated DHDK12 cells after treatment with GCV.</p> <p>Conclusions</p> <p>Our findings indicate that MTX-mediated synchronization of target cells allowed a significant improvement of retroviral HSV-<it>tk </it>gene transfer, resulting in an increased cell apoptosis in response to GCV. Pharmacological control of cell cycle may thus be a useful strategy to optimize the efficiency of retroviral-mediated cancer gene therapy.</p>