Synergistic effects of retrovirus IFN-α gene transfer and gemcitabine on apoptosis of pancreatic cancer cells
2001
adenocarcinoma cells more efficiently than wild type adenovirus. Methods: Normal esophageal squamous cells and adenocarcinoma cells were isolated from fresh surgical resection specimens and cultured in DMEM/10% FCS/PenStrep/Fungizone. After 48 hours cells were infected with the parental edenovirus or a genetically retargeted adenovitus, both encoding for the reporter gene luciferase. (AdCMVluc and AdCMVlucRGD resp.) Upon another 24 hours in culture, cells were homogenized and luciferase activity was measured in a luminometar, To estimate infection percentages, similar experiments were performed using an adenoviral vector encoding for Green Fluorescent Protein (GFP), which is detectable Jn living cells. Results: Seven primary cultures of normal esophageal and adenocarcinoma cells were established from 10 surgical resection specimens. There was a 100 x increased gene transfer in cancer cells infected with a retargeted adenovirus compared to the parental vector, while the luciferase expression was only 10 x increased in normal cells when the retargeted vector was used. Infection with the retargeted adenovirus encoding for GFP showed strong increase in the number of positive cells, indicating that the higher luciferase expression is caused by more efficient transduction. Conclusion: This study demonstrates that a genetically altered adenovirus with the REiD-motif inserted in the tiber knob infects human esophageal carcinoma cells with significantly enhanced efficiency. The increased infection of normal esophageal cells is much less pronounced. Therefore, this more efficient and more specific retargeted adenoviral vector is expected to have a better in vivo performance compared to the vectors used so far in clinical trials.
Keywords:
- Correction
- Source
- Cite
- Save
- Machine Reading By IdeaReader
0
References
1
Citations
NaN
KQI