Mobile website
Relationship between coxsackie adenovirus receptor expression levels with ad5 transduction efficiency in tumor cells
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
To investigate the relationship between coxsackie adenovirus receptor (CAR) expression and the efficiency of adenovirus gene transfer, in an effort to provide evidence for the clinical application of adenovirus-related bio-preparations. Methods: The esophageal cancer cell lines (KYSE510, KYSE150, EC9706), cervical cancer cell line (HeLa), ovary cancer cell line (SKOV3), hepatoma cell line (HepG2), and lung cancer cell line (A549) were infected with Ad-GFP (100 MOI, 200 MOI) labeled by immunofluorescence; 48 h later we measured the Ad-GFP transduction efficiencies in the above cell lines by flow cytometric analysis. CAR expression levels in vitro were assayed by Western blotting. HeLa (3×106), A549 (3×106), SKOV3 (3×106), and EC9706 (2×106) cells were implanted subcutaneously into nude mice. When the tumor sizes reached 5-7 mm in diameter, the xenografts were injected with Ad-GFP (1×109PFU) twice with a 48 h interval. After another 48 h the mice were killed and the GFP expression were observed by fluorescence microscopy to determine the transduction efficiency. CAR expression was detected by immunohistochemistry in the xenograft tissues. Results: We found that Ad transduction efficiencies varied greatly in different tissues, even among the cells of the same tissue origin. The transduction efficiencies in A549 (92.67%), HeLa (89.31%), HepG2 (84.98%) and KYSE150 (74.59%) were higher than those in SKOV3 (30.06%), KYSE510 (27.40%), and EC9706 (1893%). Consistently, GFP expression was much higher in exografts sections resulting from HeLa and A549 than those from SKOV3 and EC9706. Increased CAR expression was predictive for more efficient gene transfer in vitro and in vivo. Conclusion: CAR expression level is closely correlated to Ad transduction efficiency, which suggests that measurement of CAR expression in tumor tissues is useful for individualized adenovirus based gene therapy in clinic.
Keywords:
Project Supported:
Clc Number:
