Differential expression of a recombinant adeno-associated virus 2 vector in human CD34+ cells and breast cancer cells
| Autor: | B. Schiedlmeier, Jürgen A. Kleinschmidt, W. J. Zeller, M.R. Veldwijk, Stefan Fruehauf |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
Genetic Vectors Green Fluorescent Proteins CD34 Antigens CD34 Breast Neoplasms Biology medicine.disease_cause Transfection Multiplicity of infection Genes Reporter medicine Tumor Cells Cultured Humans Progenitor cell Molecular Biology Adeno-associated virus Cells Cultured Recombination Genetic Hematopoietic stem cell Suicide gene Dependovirus Hematopoietic Stem Cells Molecular biology Hematopoietic Stem Cell Mobilization Haematopoiesis Luminescent Proteins medicine.anatomical_structure Molecular Medicine Female |
| Zdroj: | Cancer gene therapy. 7(4) |
| ISSN: | 0929-1903 |
| Popis: | The use of autologous hematopoietic stem cell (HSC) grafts after high-dose chemotherapy protocols may be hampered by contamination of the grafts with tumor cells. Because epithelial cells seem to be the natural hosts of adeno-associated virus 2 (AAV-2), we speculated that epithelial tumor cells in HSC grafts might be selective targets for AAV-2-based vectors. To test this hypothesis, the breast cancer cell lines T47D and MCF-7 were infected with a recombinant AAV-2 vector expressing the green fluorescent protein (GFP) gene; in addition, human CD34+ mobilized peripheral progenitor cells were infected with the same vector. At a multiplicity of infection of 100, only 1.39% +/- 0.51% CD34+ cells expressed the GFP gene whereas, 36.06% +/- 6.53% of the infected T47D cells and 41.52% +/- 3.16% of the infected MCF-7 cells expressed the transduced GFP gene. After further optimizing the transduction procedure by using higher multiplicities of infection (100-500) and preincubation of samples with the tyrosine kinase inhibitor genistein, up to 82.52% and 85.35% GFP+ T47D and MCF-7 cells, respectively, were observed. The GFP fluorescence intensity in transduced mammary tumor cells was up to 3 logs higher than that of transduced CD34+ cells. The differential expression of recombinant AAV-2 vectors in hematopoietic and epithelial tumor cells warrants further research with this vector system, including the use of suicide genes for the purging of autologous HSC grafts. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|