Noninvasive visualization of microRNA-155 in multiple kinds of tumors using a radiolabeled anti-miRNA oligonucleotide

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• 作者：Lei Kang^a ; ¹ ; Yan Huo^a ; ¹ ; Quanbo Ji^b ; ¹ ; Shiyong Fan^c ; ¹ ; Ping Yan^a ; Chunli Zhang^a ; Huan Ma^a ; Pan Hao^a ; Hongwei Sun^a ; Zhibing Zheng^c ; ^{zzbcaptain@yahoo.com.cn" class="auth_mail" title="E-mail the corresponding author} ; Xiaojie Xu^d ; ^{miraclexxj@126.com" class="auth_mail" title="E-mail the corresponding author} ; Rongfu Wang^a ; ^{rongfu_wang2003@163.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：MicroRNA-155 ; Molecular imaging ; Anti-miRNA oligonucleotide ; 99mTc ; Radiolabel
• 刊名：Nuclear Medicine and Biology
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：43
• 期：2
• 页码：171-178
• 全文大小：951 K

文摘

We investigated whether a ^99mTc radiolabeled anti-miRNA-155 oligonucleotide (AMO-155) could visualize the expression of miR-155 in multiple kinds of tumors in vivo.

Methods

AMO-155 was chemically synthesized and modified with 2′-O-methyl (2′-OMe) and phosphorothioate (PS). It was radiolabeled with ^99mTc via the conjugation with NHS-MAG3 at 5′ end. The characterization of radiolabeling and serum stability was evaluated using high performance liquid chromatography (HPLC) and agarose gel electrophoresis. The expression of C/EBPβ, one of the miR-155 target proteins, was assessed using Western blot. The cellular uptake and delivery of AMO-155 was further evaluated in tumor cells. ^99mTc-AMO-155 was tested in vivo in multiple tumor models, including miR-155 over-expressed and low-expressed tumor models. Finally, biodistribution of ^99mTc-AMO-155 was evaluated.

Results

^99mTc-AMO-155 was prepared with high yield and radiochemical purity. It showed high stability in fresh human serum for 10 h. ^99mTc-AMO-155 displayed comparable capacity as unlabeled AMO-155 to increase the expression of C/EBPβ protein in MCF-7 cells. ^99mTc-AMO-155 showed an increased radioactive uptake in MCF-7 cells after 8 h of incubation, whereas no change of ^99mTc-pertechnetate uptake was observed. Carboxyfluorescein (FAM) labeled AMO-155 had higher fluorescent delivery than Control in HeLa and HepG2 cells by confocal microscopy. In miR-155 over-expressed tumor models, ^99mTc-AMO-155 showed significantly higher tumor accumulation than ^99mTc-Control. Furthermore, ^99mTc-AMO-155 was capable of discriminating between MCF-7 and MDA-MB-231 tumors based on their expression of miR-155.

Conclusions

Our study successfully prepared and proved ^99mTc-AMO-155 as a prospective imaging agent for the noninvasive visualization of miR-155 expression in vivo.