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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.

我们建立了一种新的方法，利用2- 5腺苷酸连接的针对人端粒酶RNA组分（hTR）的反义DNA抑制癌细胞端粒酶活性。该反义DNA能有效地阻断宫颈癌ME 180细胞端粒酶活性。2-5A anti-hTR对ME 180细胞的体外生长有明显的抑制作用。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。端粒长度在这一短时期内没有缩短。这些发现表明，端粒酶的封锁导致细胞生长抑制通过端粒非依赖性机制。我们证实了用2-5A抗hTR处理的细胞生长抑制是由于诱导细胞凋亡。2-5A anti-hTR诱导细胞凋亡的机制还在进一步研究中，我们还建立了新型的肿瘤基因治疗载体系统。我们以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，我们将该启动子用于基因递送系统中的癌症特异性基因表达。多种凋亡诱导基因，如caspase-8和FADD，与hTERT启动子结合，并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但周围正常组织的凋亡。