抗肿瘤多药抗性核酶转录质粒的构建及其体外切割研究

目的 探讨体外核酶对肿瘤多药抗性相关基因mdr1靶RNA分子的切割作用及其影响因素。方法 利用计算机软件设计能特异切割mdr1 mRNA的锤头状核酶（ribozyme）,并构建抗mdr1-ribozyme和靶分子的体外转录质粒,进行体外切割实验。结果 发现抗mdr1-ribozyme能较好地切割mdr1 mRNA。 结论 抗肿瘤多药抗性核酶的体外研究为其体内应用提供了参考,ribozyme有可能成为逆转肿瘤多药抗性的新药物。

The construction of a plasmid transcribing anti-mdr1 ribozyme and its cleaving reaction in vitro

Objective To study the in vitro cleavage of mdr1 target RNA by ribozyme and its influencing factors. Methods By using the computer software we designed a specific hammerhead ribozyme possessing catalytic activity that cleaves the mdr1 mRNA. A RNA-trimming plasmid pRG5mR3 and a target RNA trascribing plasmid pSABl were constructed, and a specific ribozyme cleaving reaction to target RNA was analysed. Results It was found that anti-mdr1-ribozyme is able to cleave a transcribed mdr1 mRNA fragment under physiological conditions. Conclusion The ribozyme may be targeted to inhibit the expression of the mdr1 gene in vivo, and may potentially offer a help in the strategy of cancer treatment.