Yokoyama Y, Takahashi Y, Shinohara A, Wan X, Takahashi S, Niwa K, Tamaya T
Department of Obstetrics and Gynecology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu, 500-8705, Japan.
Biochem Biophys Res Commun. 2000 Jun 24;273(1):316-21. doi: 10.1006/bbrc.2000.2939.
Because the expression level of hTERT, a catalytic subunit of human telomerase, is a rate-limiting determinant of telomerase activity, hTERT mRNA would be an excellent target of hammerhead ribozymes for the regulation of telomerase activity. We studied the efficiency of several hammerhead ribozymes targeting hTERT mRNA by transient and stable transfection procedures. To screen the potency of the ribozymes, transient ribozyme transfection and telomerase determination were performed. The ribozyme targeting 13 nucleotides downstream from the 5'-end of hTERT mRNA (13-ribozyme) exhibited the strongest telomerase-inhibitory activity, and the ribozyme to target 59 nucleotides upstream from the poly(A) tail showed clear activity. A stable transfection study confirmed that the 13-ribozyme suppressed telomerase. These observations suggest that the 13-ribozyme can regulate telomerase activity and may possess potential for cancer therapy.
由于人端粒酶的催化亚基hTERT的表达水平是端粒酶活性的限速决定因素，因此hTERT mRNA将成为锤头状核酶调节端粒酶活性的理想靶点。我们通过瞬时转染和稳定转染程序研究了几种靶向hTERT mRNA的锤头状核酶的效率。为了筛选核酶的效力，进行了瞬时核酶转染和端粒酶测定。靶向hTERT mRNA 5'-末端下游13个核苷酸的核酶（13-核酶）表现出最强的端粒酶抑制活性，而靶向多聚（A）尾上游59个核苷酸的核酶显示出明显的活性。稳定转染研究证实13-核酶可抑制端粒酶。这些观察结果表明，13-核酶可以调节端粒酶活性，并可能具有癌症治疗的潜力。
