Development of ribozymes that target stathmin, a major regulator of the mitotic spindle.
Development of ribozymes that target stathmin, a major regulator of the mitotic spindle.
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开发针对有丝分裂纺锤体主要调节因子 Stathmin 的核酶。
DOI：
10.1089/108729001750072119
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Atweh,GF
中科院分区：
文献类型：
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作者：
Mistry,SJ;Benham,CJ;Atweh,GF
Stathmin是一种主要的胞浆磷蛋白，通过调节构成有丝分裂纺锤体的微管的动力学，在控制细胞增殖中起重要作用。因为 stathmin在所有人类癌症中以高水平表达，它是抗癌干预的有吸引力的分子靶标。我们先前已经表明，反义stathmin抑制导致显著的 体外和体内白血病细胞转化表型的消除。与反义方法不同，核酶可以催化切割靶RNA的几个分子。这可能 提供了一种更有效的策略，用于下调在癌细胞中以非常高的水平表达的基因，如stathmin。我们设计了几种抗stathmin锤头状核酶，并测试了它们的活性。 对短的合成stathmin RNA底物的切割活性。体外切割研究表明stathmin RNA的位点特异性切割依赖于核酶浓度和持续时间 暴露在核酶中的风险最有活性的抗stathmin核酶能够以催化方式切割>90%的stathmin RNA，每个核酶分子切割多个底物分子。我们还证明 所设计的抗stathmin核酶能够选择性地切割从白血病细胞分离的总RNA混合物中的天然stathmin RNA。这些抗凝血酶核酶可能提供一种新的 和有效形式的基因治疗，可应用于多种人类癌症。
Stathmin is a major cytosolic phosphoprotein that plays an important role in the control of cellular proliferation by regulating the dynamics of the microtubules that make up the mitotic spindle. Because stathmin is expressed at high levels in all human cancers, it is an attractive molecular target for anticancer interventions. We had shown previously that antisense stathmin inhibition results in marked abrogation of the transformed phenotype of leukemic cellsin vitroandin vivo. Unlike the antisense approach, ribozymes can catalytically cleave several molecules of target RNA. This may provide a more efficient strategy for downregulating genes, such as stathmin, that are expressed at very high levels in cancer cells. We designed several antistathmin hammerhead ribozymes and tested their cleavage activity against short synthetic stathmin RNA substrates.In vitrocleavage studies demonstrated site-specific cleavage of stathmin RNA that was dependent on ribozyme concentration and duration of exposure to ribozyme. The most active antistathmin ribozyme was capable of cleaving >90% stathmin RNA in a catalytic manner, cleaving multiple substrate molecules per ribozyme molecule. We also demonstrated that the designed antistathmin ribozymes are capable of selectively cleaving native stathmin RNA in a mixture of total RNA isolated from leukemic cells. These antistathmin ribozymes may provide a novel and effective form of gene therapy that may be applicable to a wide variety of human cancers.