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学科主题: 临床医学
题名:
Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA
作者: Kang, Lei1; Xu, Xiao-jie2; Ma, Chao1; Wang, Rong-fu1; Yan, Ping1; Zhang, Chun-li1; Sun, Hong-wei1; Li, Ding3
关键词: Technium-99m ; MicroRNA ; Radiolabel ; Anti-miRNA oligonucleotide (AMO)
刊名: CELL BIOCHEMISTRY AND BIOPHYSICS
发表日期: 2015-03-01
DOI: 10.1007/s12013-014-0281-1
卷: 71, 期:2, 页:905-912
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biophysics ; Cell Biology
研究领域[WOS]: Biochemistry & Molecular Biology ; Biophysics ; Cell Biology
关键词[WOS]: IN-VIVO ; TC-99M ; TARGETS ; RNA ; RADIOPHARMACEUTICALS ; OLIGONUCLEOTIDES ; OLIGOMERS ; DELIVERY ; TUMORS
英文摘要:

Antisense oligonucleotides have been used for a variety of purposes in microRNA (miRNA) research including functional evaluation, target recognition, and gene studies. Although several Tc-99m-radiolabeled oligonucleotides have been reported in antisense imaging, none of those were related to miRNA tracing. Moreover, separation after labeling was always required to achieve acceptable radiochemical purity. In this study, we prepared a Tc-99m-radiolabeled oligonucleotide under optimized conditions for the purpose of tracing miRNA. A 22mer anti-miRNA oligonucleotide (AMO) was designed completely complementary to the sequence of mature miR-21. AMO probe modified with 2′-O-Methyl and phosphorothioate backbone was designed and synthesized. This probe was conjugated with a bifunctional chelator S-acetylmercaptoacetyltriglycine (NHS-MAG3) via a primary amine on 5′-end. Furthermore, it was radiolabeled and its optimization labeling conditions were performed by varying the amount of stannous ion, Tc-99m-pertecnetate, and reaction time, respectively. Finally, the labeled product was identified by gel electrophoresis and evaluated for its serum stability. The AMO was synthesized with partial 2′-OMe and phosphorothioate modification to improve its stability. Excess of MAG3 impurity was removed by precipitation of tin and MAG3 after the conjugation. The labeling efficiency reached 97 % under the optimal reaction conditions of 2 mu g/mu L SnCl2 center dot 2H(2)O addition, Tc-99m solution with high specific activity, and 90-min reaction at room temperature. Gel electrophoresis confirmed that the peak of radioactivity located the same position of oligomer, which identified the successful radiolabeling. After incubated with human fresh serum for 12 h, labeled AMO showed good stability with high radiochemical purity and no significant degradation. A Tc-99m-labeled AMO targeting miR-21 can be prepared with high labeling efficiency under optimized conditions, which provides a good support for the future use of miRNA-targeted tracing and imaging.

语种: 英语
所属项目编号: NSFC 81101065 ; 31100604 ; 81441051 ; 81472589 ; 2011YQ03011409 ; 20110001120043 ; Z141107002514159 ; Z141102001814055
项目资助者: National Natural Science Foundation of China ; National Major Scientific Equipment Special Fund ; Higher Education Doctoral Program of China ; Beijing Capital Special Development Application Program ; Beijing Nova Program
WOS记录号: WOS:000349241400050
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/65950
Appears in Collections:北京大学第一临床医学院_核医学科_期刊论文

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作者单位: 1.Peking Univ, Hosp 1, Dept Nucl Med, Beijing 100034, Peoples R China
2.Beijing Inst Biotechnol, Dept Med Mol Biol, Beijing 100850, Peoples R China
3.261 Hosp PLA, Ctr Biol Diag & Therapy, Beijing 100094, Peoples R China

Recommended Citation:
Kang, Lei,Xu, Xiao-jie,Ma, Chao,et al. Optimized Preparation of a Tc-99m-Radiolabeled Probe for Tracing MicroRNA[J]. CELL BIOCHEMISTRY AND BIOPHYSICS,2015,71(2):905-912.
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