Establishment of novel gene therapy targeting gynecologic tumors

针对妇科肿瘤的新型基因疗法的建立

基本信息

批准号：
13557138
负责人：
KYO Satoru
金额：
$ 6.72万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13557138/
关键词：
Telomerase Gene therapy hTERT hTR Gynecologic tumors 癌

项目摘要

We established novel methods that inhibit telomerase activity in cancer cells using 2-5Adenylate-linked antisense DNA against human telomerase RNA component (hTR). This antisense DNA effectively blocked telomerase activity in cervical cancer ME180 cells. Growth of ME180 cells in vitro was significantly inhibited by the treatment with 2-5A anti-hTR. Surprisingly, inhibition of cell growth was observed in 24-48 hr after treatment, quite earlier than expected. Telomere length was not shortened in this short period. These findings suggest that blockade of telomerase led to cell growth inhibition via telomere-independent mechanisms. We confirmed that growth inhibition of cells by the treatment with 2-5A anti-hTR was due to induction of apoptosis. The further anaylsis of mechanisms how 2-5A anti-hTR induces apoptosis is on going.We also established novel vector system for cancer gene therapy. We previously cloned promoter of human telomerase reverse transcriptase (hTERT), which is highly specific to cancer cells. We thus used this promoter for cancer-specific gene expression in gene delivery system. Various apoptosis-inducible genes, such as caspase-8 and FADD, were combined with hTERT promoter and used as vectors for cancer gene therapy. Introduction of these vectors effectively induced apoptosis of cancer cells but of surrounding normal tissues.

我们建立了使用 2-5 腺苷酸连接的针对人端粒酶 RNA 成分 (hTR) 的反义 DNA 抑制癌细胞中端粒酶活性的新方法。这种反义DNA有效阻断了宫颈癌ME180细胞中的端粒酶活性。 2-5A抗hTR处理显着抑制ME180细胞的体外生长。令人惊讶的是，在处理后 24-48 小时内观察到细胞生长的抑制，比预期的要早得多。端粒长度在这短暂的时间内并没有缩短。这些发现表明，端粒酶的阻断通过不依赖端粒的机制导致细胞生长抑制。我们证实用2-5A抗hTR处理对细胞生长的抑制是由于诱导细胞凋亡。 2-5A抗hTR诱导细胞凋亡机制的进一步分析正在进行中。我们还建立了用于癌症基因治疗的新型载体系统。我们之前克隆了人端粒酶逆转录酶（hTERT）的启动子，它对癌细胞具有高度特异性。因此，我们使用该启动子在基因递送系统中进行癌症特异性基因表达。各种凋亡诱导基因，如caspase-8和FADD，与hTERT启动子结合，用作癌症基因治疗的载体。这些载体的引入有效地诱导了癌细胞的凋亡，但也诱导了周围正常组织的凋亡。