Therapeutic effects of cytokine gene therapy and suicide gene therapy for gastrointestinal tumors

细胞因子基因治疗和自杀基因治疗对胃肠道肿瘤的治疗效果

基本信息

批准号：
11670556
负责人：
KAWAMURA Kiyoko
金额：
$ 1.98万
依托单位：
Chiba Cancer Center Research Institute
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-11670556/
关键词：
Gene therapy Cyotokine Suicide gene Transcriptional regulation IL-12 IL-15 IL-18 HSV-TK レトロウイルス

项目摘要

(1)We explored possibility of gene therapy for gastrointestinal tumors using cytokine genes that direct Th-1 type T cells. Three cytokine genes, IL-12, IL-15 and IL-18 gene, were cloned with a RT-PCR method. These cytokines induces the differentiation of immature T cells into Th-1 type cells or are secreted from Th-1 type cells. Human pancreatic and murine colon carcinoma cells were retrovirally transduced with the cytokine genes and were inoculated into immunocompetent or immunocompromized mice. Syngeneic mice rejected the cytokine producers and the mice developed tumor-specific, T cell-dependent protective immunity. Anti-tumor effects were also produced in immunocompromized hosts and a number of mechanisms such as anti-angiogenesis were involved in the anti-tumor activity. (2)We took another approarch to express a suicide gene specifically in tumor cells. The promoter region of the midkine gene, whose expression was predominantly observed in a number of gastrointestinal tissues but not in normal tissues, should enable the tumor-specific gene expression. In fact we detected the midkine gene expression in 14 out 15 hepatocellular specimens and identified a 0.5-kb promoter region within the regulatory sequences, which was responsible for preferential expression of a fused foreign gene in tumor cells. Human pancreatic carcinoma cells transduced with the promoter-linked herpes simplex virus-thymidine kinase gene became sensitive to ganciclovir compared with untransduced parent cells.

（1）我们使用指向T型T细胞的细胞因子基因探索了基因治疗对胃肠道肿瘤的可能性。用RT-PCR方法克隆了三个细胞因子基因IL-12，IL-15和IL-18基因。这些细胞因子诱导未成熟的T细胞分化为Th-1型细胞，或者从TH-1型细胞中分泌。用细胞因子基因逆转录病毒转导的人胰腺和鼠结肠癌细胞，并接种到免疫功能或免疫功能低下的小鼠中。合成小鼠拒绝细胞因子生产者，而小鼠产生了肿瘤特异性，T细胞依赖性保护性免疫。在免疫功能低下的宿主中还产生了抗肿瘤作用，抗肿瘤活性涉及许多机制，例如抗血管生成。（2）我们采取了另一种批准来表达在肿瘤细胞中特别表达自杀基因。 Midkine基因的启动子区域，其表达主要在许多胃肠道组织中观察到，但在正常组织中不应启用肿瘤特异性基因表达。实际上，我们检测到14个肝细胞标本中的14个中胺基因表达，并在调节序列中鉴定出一个0.5 kb的启动子区域，该序列负责在肿瘤细胞中优先表达融合的外源基因。与未转移的亲本细胞相比，用启动子连接的单纯胸腺胰岛素激酶基因转导的人胰腺癌细胞对Ganciclovir敏感。