tailieunhanh - báo cáo khoa học: "Improved retroviral suicide gene transfer in colon cancer cell lines after cell synchronization with methotrexate"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Improved retroviral suicide gene transfer in colon cancer cell lines after cell synchronization with methotrexate | Finzi et al. Journal of Experimental Clinical Cancer Research 2011 30 92 http content 30 1 92 Journal of Experimental Clinical Cancer Research RESEARCH Open Access Improved retroviral suicide gene transfer in colon cancer cell lines after cell synchronization with methotrexate 1 3 1 P s z 1 1 Laetitia Finzi Aurore Kraemer Claude Capron Severine Noullet Diane Goere Christophe Penna Bernard Nordlingen1 Josette Legagneux4 Jean-Fanpois Emile2 and Robert Malafosse1 2 Abstract Background 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-tk in two colon cancer cell lines DHDK12 and HT29. Methods 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. Results 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 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. Conclusions Our findings indicate that MTX-mediated synchronization of target cells allowed a significant improvement of retroviral HSV-fk 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