Adenovirus-mediated delivery of bFGF small interfering RNA increases levels of connexin 43 in the glioma cell line, U251
Open Access
- 14 January 2010
- journal article
- Published by Springer Science and Business Media LLC in Journal of Experimental & Clinical Cancer Research
- Vol. 29 (1), 3-7
- https://doi.org/10.1186/1756-9966-29-3
Abstract
Background bFGF is an important growth factor for glioma cell proliferation and invasion, while connexin 43 is implicated in the suppression of glioma growth. Correspondingly, gliomas have been shown to have reduced, or compromised, connexin 43 expression. Methods In this study, a bFGF-targeted siRNA was delivered to the glioma cell line, U251, using adenovirus (Ad-bFGF-siRNA) and the expression of connexin 43 and its phosphorylation state were evaluated. U251 cells were infected with Ad-bFGF-siRNA (100, 50, or 25 MOI), and infection with adenovirus expressing green fluorescent protein (Ad-GFP) at 100 MOI served as a control. Western blotting and immunofluorescence were used to detect the expression levels, phosphorylation, and localization of connexin 43 in U251 cells infected, and not infected, with Ad-bFGF-siRNA. Results Significantly higher levels of connexin 43 were detected in U251 cells infected with Ad-bFGF-siRNA at 100 and 50 MOI than in cells infected with Ad-GFP, and the same amount of connexin 43 was detected in Ad-GFP-infected and uninfected U251 cells. Connexin 43 phosphorylation did not differ between Ad-bFGF-siRNA-infected and uninfected U251 cells. However, the ratio of phosphorylated to unphosphorylated connexin 43 in Ad-bFGF-siRNA cells was lower, and connexin 43 was predominantly localized to the cytoplasm. Using a scrape loading dye transfer assay, more Lucifer Yellow was transferred to neighboring cells in the Ad-bFGF-siRNA treated group than in the control group. Conclusion To our knowledge, this is the first description of a role for connexin 43 in the inhibition of U251 growth using Ad-bFGF-siRNA.Keywords
This publication has 32 references indexed in Scilit:
- Aberrant expression and localization of connexin43 and connexin30 in a rat glioma cell lineMolecular Carcinogenesis, 2007
- The fibroblast growth factor-2 antisense gene inhibits nuclear accumulation of FGF-2 and delays cell cycle progression in C6 glioma cellsMolecular and Cellular Endocrinology, 2007
- Connexin Hemichannels and Gap Junction Channels Are Differentially Influenced by Lipopolysaccharide and Basic Fibroblast Growth FactorMolecular Biology of the Cell, 2007
- Down-regulation of Cx43 by Retroviral Delivery of Small Interfering RNA Promotes an Aggressive Breast Cancer Cell PhenotypeCancer Research, 2005
- Nuclear accumulation of basic fibroblast growth factor in human astrocytic tumorsCancer, 2003
- Beyond the gap: functions of unpaired connexon channelsNature Reviews Molecular Cell Biology, 2003
- Connexin 43–mediated bystander effect in two rat glioma cell modelsCancer Gene Therapy, 2002
- Connexin 43 (cx43) enhances chemotherapy-induced apoptosis in human glioblastoma cellsInternational Journal of Cancer, 2001
- Fibrolast growth factors in the nervous systemJournal of Neurobiology, 1994
- Gap junctions in cultured astrocytes: Single-channel currents and characterization of channel-forming proteinNeuron, 1991