Dataset Information

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

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 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.

Conclusions

Our findings indicate that MTX-mediated synchronization of target cells allowed a significant improvement of retroviral HSV-tk 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.

SUBMITTER: Finzi L

PROVIDER: S-EPMC3199255 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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Publications

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

Finzi Laetitia L Kraemer Aurore A Capron Claude C Noullet Severine S Goere Diane D Penna Christophe C Nordlinger Bernard B Legagneux Josette J Emile Jean-Fançois JF Malafosse Robert R

Journal of experimental & clinical cancer research : CR 20111004

<h4>Background</h4>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 thymidi ...[more]

PMID: 21970612

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Dataset Information

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

Background

Methods

Results

Conclusions

Publications

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

Similar Datasets

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