Home Neoplasma 2006 Neoplasma Vol.53, p.353-362, 2006

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Founded: 1954
ISSN 0028-2685
ISSN 1338-4317 (online)

Published in English

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Neoplasma Vol.53, p.353-362, 2006

Title: Combine cancer gene therapy harnessing plasmids expressing human tumor necrosis factor alpha and Herpes simplex thymidine kinase suicide gene

Abstract: We have assessed the effect of combine cancer gene therapy with exogenous human tumor necrosis factor alpha (hTNFalpha) and suicide gene therapy on three human cancer cell lines MCF-7 (breast adenocarcinoma), U-118MGand 42-MG-BA (human gliomas). Transfection of a plasmid containing hTNFalpha under the control of a hybrid promoter resulted in expression of hTNFalpha gene in vitro. Transduction of retroviral plasmid containing Herpes simplex thymidine kinase (HSVtk) led to the expression of thymidine kinase in all three cell lines. MTT cell proliferation assay and flow cytometric analysis showed a significant increase in apoptotic and necrotic cells and decrease of proliferation in all cell lines after combine therapy with hTNFalpha expression plus thymidine kinase/GCV suicide system. The presence of these two genes after transduction of retroviral vector containing thymidine kinase and hTNFalpha was confirmed by PCR. The expression of HSVtk gene was proved by Western blot analysis, and the expression of both genes was confirmed by RT-PCR. Additive cell killing effect due to presence of HSVtk and hTNFalpha therapeutic genes after activation of non-toxic prodrug was observed. Whether the bicistronic plasmid containing both genes would improve the therapeutic effect need to be assessed in the future.

Keywords: cancer gene therapy, retroviral vector, Herpes simplex thymidine kinase, tumor necrosis factor alpha, apoptosis
Year: 2006, Volume: 53, Issue: Page From: 353, Page To: 362

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