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Neoplasma Vol.53, p.353-362, 2006 |
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Title: Combine cancer gene therapy harnessing plasmids expressing human tumor necrosis factor alpha and Herpes simplex thymidine kinase suicide gene | ||
Author: A., PASTORAKOVA ; K., HLUBINOVA ; J., JAKUBIKOVA ; C., ALTANER ; | ||
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.
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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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