| 马 超,康 磊,王荣福.MicroRNA靶向的放射性标记探针的制备及条件优化[J].肿瘤学杂志,2013,19(12):918-924. |
| MicroRNA靶向的放射性标记探针的制备及条件优化 |
| Preparation and Optimization of 99mTc-radiolabeled Antisense Oligonucleotide Targeting microRNA |
| 投稿时间：2013-11-20 |
| DOI：10.11735/j.issn.1671-170X.2013.12.B003 |
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| 中文关键词: 锝［99mTc］ microRNA（miRNA） 放射性标记 |
| 英文关键词:technium-99m microRNA radiolabeling |
| 基金项目:国家自然科学基金（81101065、81071183、31100604）；国家重大科学仪器设备开发专项（2011YQ03011409）；高等学校博士学科点新教师专项科研基金（20110001120043） |
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| 中文摘要: |
| 摘 要：［目的］ 以microRNA-21（miR-21）为靶点，探索制备放射性核素锝［99mTc］标记的反义寡核苷酸探针的方法与实验条件。［方法］设计化学结构修饰的反义寡核苷酸序列并进行合成，利用双功能螯合剂NHS-MAG3偶联待标记探针和99mTc，优化不同实验条件下的标记率及放化纯，并对标记产物进行凝胶电泳鉴定及稳定性评价。［结果］ 降低氯化亚锡的用量、减少99mTc的反应体积、适度延长标记反应时间均可显著提高标记率，最高可达97%。标记探针在血清37℃条件中孵育6h其放化纯仍可维持在96%以上。凝胶电泳结果证实核酸条带与放射性分布一致。［结论］通过优化探针设计和标记反应条件能够大大提高标记率和放化纯度，无需纯化即可进一步应用，达到简化制备程序的目的。 |
| 英文摘要: |
| Abstract：［Purpose］ To explore the methods and optimized reaction conditions for the preparation of 99mTc radiolabeled antisense oligonucleotide targeted microRNA-21. ［Methods］ The antisense oligonucleotide probe with chemical and structural modifications was designed and synthesized. This probe was radiolabeled with 99mTc via the conjugation of a bifunctional chelator S-acetylmercaptoacetyltriglycine (NHS-MAG3). The labeled probe was tested for its radiolabeling efficiency，purity and its stability under different reaction conditions，as well as the identification with gel electrophoresis. ［Results］ The labeling efficiency reached to 97% under optimized reaction conditions of different volume of SnCl2·2H2O，99mTc and reaction time. The radiochemical purity of 99mTc-anti-miR in fresh human serum at 37℃ could maintained more than 96% after 6h. ［Conclusions］ The labeling efficiency can be improved greatly by optimizing design and reaction conditions. Therefore，the probe could be prepared easily and simply for further applications without further purification. |
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