Adenovirus-mediated suicide gene therapy using the human telomerase catalytic subunit (hTERT) gene promoter induced apoptosis of ovarian cancer cell line
Adenovirus-mediated suicide gene therapy using the human telomerase catalytic subunit (hTERT) gene promoter induced apoptosis of ovarian cancer cell line
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DOI：
10.1271/bbb.67.2344
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发表时间：
2003-11-01
影响因子：
1.6
通讯作者：
Kim, YT
中科院分区：
文献类型：
--
作者：
Song, JS;Kim, HP;Kim, YT
端粒酶是一种核糖核蛋白复合物，其功能是将端粒重复序列（TTAGGG）（n）添加到染色体末端，已知它在细胞永生化中发挥重要作用。端粒酶在大多数肿瘤细胞中高度活跃，但在正常细胞中则不然。因此，它可能在癌症基因治疗中具有应用前景。端粒酶由两个基本成分组成：端粒酶RNA模板（hTR）和催化亚基（hTERT）。 hTERT 仅在端粒酶活性呈阳性的细胞和组织中表达，即肿瘤和胎儿细胞。我们在此测试了在靶向癌症基因治疗中利用 hTERT 启动子的可能性。我们在腺病毒穿梭质粒中克隆了hTERT启动子来替代CMV启动子，并亚克隆了HSV-TK基因以受hTERT基因启动子控制。然后我们构建重组腺病毒Ad-hT-TK，并将其感染正常和人妇科癌细胞系。通过这些实验，我们鉴定了 Ad-hT-TK 选择性肿瘤特异性细胞死亡。此外，FACS 分析和 TUNEL 测定表明，活力的降低是通过诱导细胞凋亡介导的，表明这种方法可能是靶向癌症基因治疗中抑制癌症生长的有用方法。这些结果表明Ad-hT-TK可用于妇科癌症基因治疗。
Telomerase is a ribonucleoprotein complex the function of which is to add telomeric repeats (TTAGGG)(n) to chromosomal ends, and it is known to play an important role in cellular immortalization. Telomerase is highly active in most tumor cells, yet not in normal cells. As such, it may have possible applications in cancer gene therapy. Telomerase consists of two essential components, telomerase RNA template (hTR) and catalytic subunit (hTERT). hTERT is expressed only in cells and tissues positive for telomerase activity, i.e., tumor and fetal cells. We here tested the possibility of the utilization of the hTERT promoter in targeted cancer gene therapy. We cloned the hTERT promoter in the replace of the CMV promoter and sub-cloned HSV-TK gene to be controlled by hTERT gene promoter in adenovirus shuttle plasmid. Then we constructed recombinant adenovirus Ad-hT-TK, and infected them into normal and human gynecological cancer cell lines. Through these experiments, we identified the selective tumor specific cell death by Ad-hT-TK. Furthermore, FACS analysis and TUNEL assay suggests that the reduced viability is mediated through the induction of apoptosis, indicating that this approach may be a useful method for suppressing cancer growth in targeted cancer gene therapy. These results show that Ad-hT-TK could be used for gynecological cancer gene therapy.