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1
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Recombinant angioarrestin secreted from mouse melanoma cells inhibits growth of primary tumours

100%
Smagur A., Szary J., Szala S.
Acta Biochimica Polonica
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2005
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vol. 52
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issue 4
875-879
EN
Angioarrestin is a recently described anti-angiogenic protein whose expression is down-regulated in solid tumours of various origins. It has a sequence identical to angiopoietin related protein-1. In this study we investigated anti-tumour properties of angioarrestin in B16 (F10) melanoma tumour model. We constructed an expression vector encoding human angioarrestin under the control of EF-1α promoter. This vector was transferred to B16 (F10) cells and recombinant angioarrestin secreted from the transfected cells was tested for anti-angiogenic activity using endothelial cell proliferation assay. Finally, mice were injected subcutaneously with cells that had been transfected with either angioarrestin-encoding vector or empty vector and tumor growth was compared. The obtained recombinant angioarrestin inhibited proliferation of bovine aortic endothelial cells. Tumours derived from an angioarrestin-secreting B16 (F10) cell clone grew in vivo more slowly than tumours derived from a cell clone transfected with empty vector. These data show, to our knowledge for the first time, that angioarrestin can inhibit primary melanoma tumour growth.
2
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Direct transfer of IL-12 gene into growing Renca tumors.

100%
Budryk M., Wilczyńska U., Szary J., Szala S.
Acta Biochimica Polonica
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2000
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vol. 47
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issue 2
385-391
EN
We investigated the feasibility of transferring naked plasmid DNA containing a therapeutic gene (IL-12) into mice harboring growing Renca tumors. We found that naked DNA transferred into growing Renca and B16(F10) tumors gives higher expression level of reporter gene than complexes of DNA with DDAB/ DOPE or DC-Chol/DOPE. Transfer of naked DNA carrying the IL-12 gene into growing Renca tumors causes a distinct therapeutic effect that depends on the time span between inoculation of mice with cancer cells and the beginning of the therapy. Therapy started on day 3 resulted in total cure (100%) of mice.
3
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Combined delivery of an antiangiogenic protein (angiostatin) and an immunomodulatory gene (interleukin-12) in the treatment of murine cancer.

100%
Wilczyńska U., Kucharska A., Szary J., Szala S.
Acta Biochimica Polonica
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2001
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vol. 48
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issue 4
1077-1084
EN
We investigated the feasibility of a novel therapeutic approach to treat neoplastic diseases in mice. This novel strategy consists in delivering a protein (angiostatin) with strong antiangiogenic properties, followed by administration of the interleukin 12 gene that is strongly immunomodulatory and has also some antiangiogenic effects. When angiostatin-mediated antiangiogenic therapy was used in combination with intratumor delivery of the IL-12 gene (a strategy much safer than IL-12 protein administration), this produced a synergistic therapeutic effect.
4
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Antiangiogenic gene therapy in inhibition of metastasis.

100%
Szala S., Szary J., Cichoń T., Sochanik A.
Acta Biochimica Polonica
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2002
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vol. 49
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issue 2
313-321
EN
This short review attempts to demonstrate the usefulness of antiangiogenic gene therapy in achieving inhibition of growth in experimentally-induced metastases. Certain normal tissues (for example skeletal muscle) may be used in vivo, after genetic modification, as a "bioreactor", able to produce and secrete into the bloodstream proteins known to exert antiangiogenic effects. By inhibiting neoangiogenesis these proteins would thus prevent the development of metastases. The review discusses also the perspectives of antimetastatic therapy based on certain types of allogenic cells (for example myoblasts and fibroblasts) that had been genetically modified and then microencapsulated. The strategy of encapsulation is aimed at protecting the modified cells secreting antiangiogenic factors from being eliminated by the immune system. Secretion of antiangiogenic proteins by these microencapsulated cells can be controlled with inducible promoters. Antiangiogenic genes remaining under the transcriptional control of such promoters may be switched on and off using antibiotics, such as tetracycline derivatives, or steroid hormones.
5
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Inhibition of tumor growth by interleukin 10 gene transfer in B16(F10) melanoma cells

100%
Walos S., Szary J., Szala S.
Acta Biochimica Polonica
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1999
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vol. 46
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issue 4
967-970
EN
Interleukin 10 (IL-10) is a potent immunosuppressive cytokine with an antitumor activity. The effect of IL-10 on tumor growth was tested in murine melanoma cells manipulated by gene transfer to secrete IL-10. In mice bearing B16(F10) tumors expressing IL-10  tumor growth was decreased depending on the amount of secreted IL-10.
6
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CEA-negative glioblastoma and melanoma cells are sensitive to cytosine deaminase/5-fluorocytosine therapy directed by the carcinoembryonic antigen promoter.

88%
Dąbrowska A., Szary J., Kowalczuk M., Szala S., Ugorski M.
Acta Biochimica Polonica
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2004
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vol. 51
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issue 3
723-732
EN
Recent studies have suggested that carcinoembryonic antigen (CEA)-promoter sequences are active only in CEA-positive cells, filing in the criteria for tumor specific targeting of suicide genes. However, the present study on gene therapy of colon cancer and cell-specificity of CEA promoter, provide evidence that CEA-positive and CEA-negative cells transfected with E. coli cytosine deaminase (CD) gene under the control of CEA promotor sequence are sensitive to enzyme/pro-drug therapy with 5-fluorocytosine (5-FC). Individual clones derived from the CEA-negative cell lines: melanoma Hs294T and glioblastoma T98G after transfection with CD differed profoundly in their sensitivity to 5-FC. The IC50 values for several clones of the CEA-negative cells were almost the same as for CEA-positive colon cancer cells. Such 5-FC-sensitive clones derived from the population of CEA-negative cells, present even in small number, because of the very effective bystender effect of this enzyme/pro-drug system can cause severe problems during therapy by efficiently killing surrounding normal cells. Safety is the major issue in gene therapy. Our data suggest that the safety of gene-directed enzyme pro-drug therapy (GDEPT) with CEA promoter driven expression of therapeutic genes is not so obvious as it has originally been claimed.
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