Gene therapy for hepatocellular carcinoma

肝细胞癌的基因治疗

• 批准号: 11670516
• 负责人: IDO Akio
• 金额: $ 0.51万
• 依托单位: Miyazaki Medical College
• 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-11670516/
• 关键词: gene therapy; hepatoma; α-fetoprotein; hypoxia; VEGF; HSV-tk gene; gancyclovir; suicide gene; 血管新生

We previously reported that the retroviral vector expressing the herpes simplex virus-thymidine kinase (HSV-tk) gene under the control of 0.3-kb human α-fetoprotein (AFP) gene promoter (AF0.3) provided the cytotoxicity to ganciclovir (GCV) in high-AFP-producing human hepatoma cells but not in low-AFP-producing cells. Therefore, specific enhancement of AFP promoter activity is likely to be required to induce enough cytotoxicity in low-AFP-producing hepatoma cells. In this study, we constructed a hybrid promoter, [HRE]AF, in which a 0.4-kb fragment of human vascular endothelial growth factor (VEGF) 5'-flanking sequences containing hypoxia responsive element (HRE) was fused to AF0.3 promoter. By means of the reporter gene transfection assay, hypoxia-inducible transcriptions that were mediated by [HRE]AF promoter were detected in low- and non-AFP-producing human hepatoma cells, but not in nonhepatoma cells. When the HSV-tk gene controlled by [HRE]AF promoter was transduced into hepatoma and nonhepatoma cells by a retroviral vector, the exposure to 1% O2 induced GCV-cytotoxicity specifically in the hepatoma cells. Moreover, in nude mice bearing solid tumor xenografts, only the tumors consisting of the virus-infected hepatoma cells gradually disappeared by GCV administration. These results indicate that the hypoxia-inducible enhancer of the human VEGF gene, which is directly linked to human AFP promoter, involves selective and enhanced tumoricidal activity in gene therapy for hepatocellular carcinoma.

我们以前报道过，在0.3kb人甲胎蛋白（AFP）基因启动子（AF0.3）控制下表达单纯疱疹病毒胸苷激酶（HSV-tk）基因的逆转录病毒载体，在高AFP产生的人肝癌细胞中对更昔洛韦（GCV）具有细胞毒作用，而在低AFP产生的细胞中则没有。因此，可能需要特异性增强AFP启动子活性以在低AFP产生的肝癌细胞中诱导足够的细胞毒性。在本研究中，我们构建了一个杂合启动子，[HRE]AF，其中含有缺氧反应元件（HRE）的人血管内皮生长因子（VEGF）5 '侧翼序列的0.4-kb片段融合到AF 0.3启动子。通过报告基因转染实验，[HRE]AF启动子介导的低氧诱导转录在低AFP产生和非AFP产生的人肝癌细胞中被检测到，而在非肝癌细胞中未检测到。将[HRE]AF启动子控制的HSV-tk基因通过逆转录病毒载体导入肝癌细胞和非肝癌细胞后，1%O_2暴露可特异性诱导肝癌细胞的GCV细胞毒活性。此外，在荷实体瘤异种移植物的裸鼠中，只有由病毒感染的肝癌细胞组成的肿瘤通过GCV给药逐渐消失。这些结果表明，缺氧诱导的增强子的人VEGF基因，这是直接连接到人AFP启动子，涉及选择性和增强的杀肿瘤活性的基因治疗肝细胞癌。

项目成果

Akio Ido et al.: "Gene therapy targeting for hepatocellular carcinoma : selective and enhanced suicide gene expression regulated by hypoxia-inducible enhancer linked to human a-fetoprotein promoter."Cancer Research. 61(in press). (2001)

Akio Ido 等人：“针对肝细胞癌的基因治疗：通过与人甲胎蛋白启动子连接的缺氧诱导增强子调节选择性和增强的自杀基因表达。”癌症研究。

Hirofumi Uto et al..: "Hepatoma-specific gene therapy through retrovirus-mediated and targeted gene transfer using an adenovirus carrying the ecotropic receptor gene."Biochem.Biophys.Res.Commun.. 265. 550-555 (1999)

Hirofumi Uto 等人：“使用携带亲嗜性受体基因的腺病毒，通过逆转录病毒介导的靶向基因转移进行肝癌特异性基因治疗。”Biochem.Biophys.Res.Commun.. 265. 550-555 (1999)

Ido A, Uto H, et al.: "Gene therapy targeting for hepatocellular carcinoma : Selective and enhanced gene expression regulated by hypoxia-inducible enhancer linked to human α-fetoprotein promoter"Cancer Research. 61 (in press). (2001)

Ido A、Uto H 等人：“针对肝细胞癌的基因治疗：通过与人 α-胎蛋白启动子连接的缺氧诱导增强子调节选择性和增强的基因表达”癌症研究 61（出版中）。

坪内博仁 他2名: "肝細胞癌の遺伝子治療-将来の展望"臨床消化器内科. 14・7. 268-278 (1999)

Hirohito Tsubouchi 等 2 人：“肝细胞癌的基因治疗 - 未来前景”临床胃肠病学 14・7（1999）。

Uto H, Ido A, Hori T, et al.: "Hepatoma-specific gene therapy through retrovirus-mediated and targeted gene transfer using an adenovirus carrying the ecotropic receptor gene"Biochem Biophys Res Commun. 265. 550-555 (1999)

Uto H、Ido A、Hori T 等人：“使用携带亲嗜性受体基因的腺病毒，通过逆转录病毒介导的靶向基因转移进行肝癌特异性基因治疗”Biochem Biophys Res Commun。