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Suicide gene therapy using adenovirus vector for human oral squamous carcinoma cell line in vitro
Noriyuki Yamamoto1, Yasushi Hayashi, Hideaki Kagami
1Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan. noriyuki@med.nagoya-u.ac.jp
Abstract:
Recently, suicide gene therapy using the herpes simplex virus thymidine kinase (HSVtk) gene followed by ganciclovir (GCV) administration was evaluated for the treatment of cancer. The purpose of this study was to investigate the effectiveness of suicide gene therapy using the replication-deficient recombinant adenovirus vector for human oral squamous carcinoma cell lines. To evaluate transduction efficiency, each cell line was transduced in vitro with an adenovirus vector containing the beta-galactosidase gene. By 24 hours after transduction, nearly 100% of the cells were transduced at a multiplicity of infection (MOI) of 10, and from 30 to 10% at an MOI of 1. Next, each cell line was transduced with an adenovirus vector containing the HSVtk gene, and a subsequent administration of GCV for the assessment of suicide gene therapy. A subsequent administration of GCV resulted in complete tumor cell death. In addition, we conducted a morphological analysis of that cell death using video-enhanced contrast differential interference contrast microscopy, and we observed that it included both apoptosis and necrosis after HSVtk gene and GCV treatment. These results suggest that adenovirus-mediated suicide gene therapy induced remarkable cytotoxicity with a bystander effect in human oral squamous cell carcinoma thus suggesting an effective treatment strategy for that tumor.
Insights
Suicide gene therapy using herpes simplex virus thymidine kinase (HSVtk) and ganciclovir (GCV) effectively killed oral cancer cells. Adenovirus vectors demonstrated high transduction efficiency, leading to apoptosis and necrosis, suggesting a promising cancer treatment.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Suicide gene therapy, utilizing the herpes simplex virus thymidine kinase (HSVtk) gene and ganciclovir (GCV), is an emerging cancer treatment.
- Adenovirus vectors are being explored for their potential in delivering therapeutic genes effectively.
Purpose of the Study:
- To evaluate the efficacy of adenovirus-mediated suicide gene therapy for human oral squamous carcinoma.
- To assess the transduction efficiency of adenovirus vectors in oral cancer cell lines.
Main Methods:
- In vitro transduction of oral squamous carcinoma cell lines with adenovirus vectors carrying the beta-galactosidase gene to determine transduction efficiency.
- Transduction with an adenovirus vector containing the HSVtk gene, followed by GCV administration.
- Morphological analysis of cell death using video-enhanced contrast differential interference contrast microscopy.
Main Results:
- Adenovirus vectors achieved high transduction efficiency (nearly 100% at MOI 10) in oral cancer cell lines.
- Subsequent GCV administration after HSVtk gene transduction resulted in complete tumor cell death.
- Morphological analysis revealed that cell death occurred through both apoptosis and necrosis.
Conclusions:
- Adenovirus-mediated suicide gene therapy with HSVtk/GCV is a potent strategy for inducing significant cancer cell death.
- The observed bystander effect contributes to the remarkable cytotoxicity, indicating its potential as an effective treatment for oral squamous cell carcinoma.
