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Transduction of normal and malignant oral epithelium by particle bombardment
E J Shillitoe1, S Noonan, C C Hinkle
1Department of Microbiology and Immunology, State University of New York College of Medicine, Syracuse 13210, USA.
Abstract:
Although genetic approaches to the treatment and prevention of oral cancer are being developed, there are no suitable methods of transduction of the oral mucosa or early cancers. We therefore tested the technique of particle bombardment for its ability to transduce oral cancer cells in vitro and normal epithelium of the hamster cheek pouch in vivo. A gene gun was used to transfer a plasmid that encoded a marker/suicide fusion gene, beta-galactosidase-thymidine kinase (GAL-TEK), under control of a CMV promoter. For comparison we used the method of lipofection and an adenovirus vector. Particle bombardment transduced up to 13% of cells in culture, resulting in a 24.3% reduction in growth in the presence of ganciclovir. The efficiency of transduction was similar to that of lipofection but was much less than that of the adenovirus vector, which transduced 54% of cells and completely inhibited their growth in the presence of ganciclovir. Transduction of the hamster cheek pouch by particle bombardment produced expression of beta-galactosidase as judged by macroscopic staining, for up to 5 days. However, histological examination showed that the transduced cells were rare and superficial, and that administration of systemic ganciclovir did not lead to any changes in the tissue. Improvements in efficiency are necessary before the gene gun can be used in the management of oral cancer.
Insights
Particle bombardment shows limited success in gene transfer for oral cancer treatment. While it transduced some cells, its efficiency is lower than adenovirus vectors, requiring further improvements for clinical application.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Genetic approaches for oral cancer treatment are emerging.
- Effective transduction methods for oral mucosa and early cancers are lacking.
- Particle bombardment (gene gun) is explored as a potential gene delivery technique.
Purpose of the Study:
- To evaluate the efficacy of particle bombardment for transducing oral cancer cells in vitro.
- To assess particle bombardment's ability to transduce normal hamster cheek pouch epithelium in vivo.
- To compare particle bombardment with lipofection and adenovirus vectors for gene delivery.
Main Methods:
- Utilized a gene gun to deliver a beta-galactosidase-thymidine kinase (GAL-TEK) fusion gene.
- Tested gene delivery in oral cancer cells in vitro and hamster cheek pouch epithelium in vivo.
- Compared transduction efficiency and therapeutic effect with lipofection and adenovirus vectors.
Main Results:
- Particle bombardment transduced up to 13% of oral cancer cells in vitro, reducing growth by 24.3% with ganciclovir.
- In vivo, particle bombardment showed beta-galactosidase expression for up to 5 days, but transduced cells were rare and superficial.
- Adenovirus vector demonstrated higher transduction efficiency (54%) and complete growth inhibition compared to particle bombardment and lipofection.
Conclusions:
- Particle bombardment shows limited efficiency for transducing oral mucosa and early oral cancers.
- Current gene gun technology requires significant improvement for effective gene therapy in oral cancer management.
- Adenovirus vectors appear more promising for oral cancer gene therapy based on this comparative study.
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