Study of the suicide gene therapy using HSVtk gene and GCV for oral cancer

HSVtk基因和GCV自杀基因治疗口腔癌的研究

• 批准号: 11671984
• 负责人: HAYASHI Yasushi
• 金额: $ 2.5万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671984/
• 关键词: Helpes simplex virus thymidinekinase gene; Ganciclovir; Suicide gene therapy; Adenovirus vector; Adeno-associated virus vector

Gene therapy offers the possibility of new approach to cancer treatment. The purpose of this study was to test the feasibility of suicide gene therapy using herpes simplex virus thymidine kinase (HSVtk) gene and ganciclovir (GCV) for oral cancer. Four human oral squamous cell carcinoma sell lines were used. To evaluate transduction efficiency, each cell line was transduced in vitro with an adenovirus vector (Ad) or adeno-associated virus vector (AAV) containing the β-galactosidase gene. By 24 hours after transduction, nearly 100% of the cells were transduced at a multiplicity of infection (MOI) of 10, and from 10 to 30% at a MOI of 1 in Ad. Transduction efficiency in AAV was nearly 100% at a MOI of 10000, and from 40 to 50% at a MOI of 1000. Next, each cell line was transduced with an Ad or AAV containing the HSVtk gene, and subsequent administration of GCV for the assessment of suicide gene therapy. Subsequent administration of GCV resulted in complete tumor cell death. Ad or AAV containing the HSVtk gene injection followed by intraperiotoneal GCV administration inhibited remarkably tumor growth in the nude mice to which oral squamous cell carcinoma cell SAS was transplanted SCID mouse. These results suggest that suicide gene therapy with HSVtk and GCV has revealed an effective treatment strategy for the oral cancer.

基因治疗为癌症治疗提供了新方法的可能性，本研究的目的是测试使用单纯疱疹病毒胸苷激酶（HSVtk）基因和更昔洛韦（GCV）治疗口腔癌的自杀基因治疗的可行性。为了评估转导效率，用含有β-半乳糖苷酶的腺病毒载体（Ad）或腺相关病毒载体（AAV）在体外转导每个细胞系。转导后 24 小时，感染复数 (MOI) 为 10 时几乎 100% 的细胞被转导，而当 MOI 为 1 时，Ad 中的转导效率接近 100%。 MOI 为 10000，MOI 为 1000 时为 40% 至 50%。接下来，用含有HSVtk基因的Ad或AAV，以及随后施用GCV以评估自杀基因治疗，随后施用含有HSVtk基因的Ad或AAV导致肿瘤细胞完全死亡，随后腹膜内施用GCV显着抑制肿瘤生长。移植口腔鳞状细胞癌细胞SAS的裸鼠SCID小鼠这些结果表明HSVtk和GCV的自杀基因治疗揭示了口腔癌的有效治疗策略。

项目成果

林康司: "ヒト口腔扁平上皮癌細胞株:HSC-4に対するアデノ随伴ウイルスベクターを用いた自殺遺伝子治療"日本口腔外科学会雑誌. 45(9). 543-546 (1999)

Koji Hayashi：“使用腺相关病毒载体对抗人口腔鳞状细胞癌细胞系：HSC-4”，日本口腔颌面外科杂志 45(9)。

Fukuhara H., Hayashi Y., Kagami H., Fukui T., Yamamoto N., Tohnai I., Ueda M., Mizuno M., Yoshida J.: "Suicide gene therapy with adenovirus vector containing the herpes simplex thymidine kinase gene for head and neck squamous cell carcinoma cell in vivo."

Fukuhara H.、Hayashi Y.、Kagami H.、Fukui T.、Yamamoto N.、Tohnai I.、Ueda M.、Mizuno M.、Yoshida J.：“使用含有单纯疱疹胸苷激酶基因的腺病毒载体进行自杀基因治疗

Fukui T., Hayashi Y., Kagami H., Yamamoto N., Fukuhara H., Tohnai I., Ueda M., Mizuno M., Yoshida J.: "Suicide gene therapy for human oral squamous cell carcinoma cell lines with adeno-associated virus vector."Euro.J.Cancer (Oral Oncology). (2001)

Fukui T.、Hayashi Y.、Kagami H.、Yamamoto N.、Fukuhara H.、Tohnai I.、Ueda M.、Mizuno M.、Yoshida J.：“用腺苷对人口腔鳞状细胞癌细胞系进行自杀基因治疗

福原広和: "口腔癌に対するアデノウイルスベクターを用いた自殺遺伝子治療"頭頚部腫瘍. 27(1). (2001)

Hirokazu Fukuhara：“使用腺病毒载体治疗口腔癌的自杀基因疗法”头颈肿瘤27（1）。