Yamamoto Noriyuki, Hayashi Yasushi, Kagami Hideaki, Fukui Takafumi, Fukuhara Hirokazu, Tohnai Iwai, Ueda Minoru, Mizuno Masaaki, Yoshida Jun
Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan.
Nagoya J Med Sci. 2005 Jun;67(3-4):83-91.
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.
最近，对使用单纯疱疹病毒胸苷激酶（HSVtk）基因并随后给予更昔洛韦（GCV）的自杀基因疗法进行了癌症治疗评估。本研究的目的是调查使用复制缺陷型重组腺病毒载体对人口腔鳞状癌细胞系进行自杀基因治疗的有效性。为了评估转导效率，每个细胞系在体外被含有β-半乳糖苷酶基因的腺病毒载体转导。转导后24小时，感染复数（MOI）为10时，近100%的细胞被转导，MOI为1时，转导率为30%至10%。接下来，每个细胞系被含有HSVtk基因的腺病毒载体转导，随后给予GCV以评估自杀基因疗法。随后给予GCV导致肿瘤细胞完全死亡。此外，我们使用视频增强对比微分干涉对比显微镜对该细胞死亡进行了形态学分析，并且我们观察到在HSVtk基因和GCV处理后，细胞死亡包括凋亡和坏死。这些结果表明，腺病毒介导的自杀基因疗法在人口腔鳞状细胞癌中诱导了具有旁观者效应的显著细胞毒性，从而提示了一种针对该肿瘤的有效治疗策略。
