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全细胞 SELEX 适体用于体内高特异性癌荧光分子成像。

Whole cell-SELEX aptamers for highly specific fluorescence molecular imaging of carcinomas in vivo.

机构信息

State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, China.

出版信息

PLoS One. 2013 Aug 12;8(8):e70476. doi: 10.1371/journal.pone.0070476. eCollection 2013.

Abstract

BACKGROUND

Carcinomas make up the majority of cancers. Their accurate and specific diagnoses are of great significance for the improvement of patients' curability.

METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we report an effectual example of the in vivo fluorescence molecular imaging of carcinomas with extremely high specificity based on whole cell-SELEX aptamers. Firstly, S6, an aptamer against A549 lung carcinoma cells, was adopted and labeled with Cy5 to serve as a molecular imaging probe. Flow cytometry assays revealed that Cy5-S6 could not only specifically label in vitro cultured A549 cells in buffer, but also successfully achieve the detection of ex vivo cultured target cells in serum. When applied to in vivo imaging, Cy5-S6 was demonstrated to possess high specificity in identifying A549 carcinoma through a systematic comparison investigation. Particularly, after Cy5-S6 was intravenously injected into nude mice which were simultaneously grafted with A549 lung carcinoma and Tca8113 tongue carcinoma, a much longer retention time of Cy5-S6 in A549 tumor was observed and a clear targeted cancer imaging result was presented. On this basis, to further promote the application to imaging other carcinomas, LS2 and ZY8, which are two aptamers selected by our group against Bel-7404 and SMMC-7721 liver carcinoma cells respectively, were tested in a similar way, both in vitro and in vivo. Results showed that these aptamers were even effective in differentiating liver carcinomas of different subtypes in the same body.

CONCLUSIONS/SIGNIFICANCE: This work might greatly advance the application of whole cell-SELEX aptamers to carcinomas-related in vivo researches.

摘要

背景

癌瘤构成了大多数癌症。准确和特异性的诊断对提高患者的治愈率具有重要意义。

方法/主要发现:在本文中,我们报告了一个基于全细胞 SELEX 适体的癌瘤高特异性体内荧光分子成像的有效实例。首先,采用针对 A549 肺癌细胞的适体 S6,并将其标记为 Cy5 用作分子成像探针。流式细胞术检测表明,Cy5-S6 不仅可以特异性标记缓冲液中体外培养的 A549 细胞,而且还可以成功地检测到血清中培养的靶细胞。当应用于体内成像时,Cy5-S6 通过系统比较研究被证明在识别 A549 癌瘤方面具有高度特异性。特别是,当 Cy5-S6 静脉注射到同时移植 A549 肺癌和 Tca8113 舌癌的裸鼠体内后,在 A549 肿瘤中观察到 Cy5-S6 更长的保留时间,并呈现出清晰的靶向癌症成像结果。在此基础上,为了进一步促进对其他癌瘤成像的应用,我们小组分别针对 Bel-7404 和 SMMC-7721 肝癌细胞选择的 LS2 和 ZY8 适体,以类似的方式在体外和体内进行了测试。结果表明,这些适体甚至可以有效地区分同一体内不同亚型的肝癌。

结论/意义:这项工作可能会极大地推动全细胞 SELEX 适体在癌瘤相关的体内研究中的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8415/3741280/56c73be3d537/pone.0070476.g001.jpg

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