Development of a novel DNA aptamer ligand targeting to primary cultured tumor endothelial cells by a cell-based SELEX method.
Development of a novel DNA aptamer ligand targeting to primary cultured tumor endothelial cells by a cell-based SELEX method.
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DOI：
10.1371/journal.pone.0050174
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Harashima H
中科院分区：
文献类型：
--
作者：
Ara MN;Hyodo M;Ohga N;Hida K;Harashima H
本研究使用自发的基于细胞的SELEX方法（通过指数富集的配体的系统进化）来产生特异性结合由原代培养的小鼠肿瘤内皮细胞（mTEC）产生的细胞表面蛋白或生物标志物的DNA适体。在实体瘤中，新血管通过血管生成过程形成，这在癌症发展和转移中起着关键作用。为了对抗血管生成，对不同癌症类型进行适当的诊断和分子水平的了解现在是一个高度优先事项。本研究开发的新型DNA适体AraHH001在纳摩尔范围内以高亲和力特异性结合mTEC，但不结合正常皮肤内皮细胞（皮肤EC）。还发现所选择的DNA适体与从患有肾癌的临床患者分离的培养的人肿瘤内皮细胞（hTEC）结合。AraHH001通过抑制基质胶上的mTECs的管形成而显示出显著的抗血管生成活性。有趣的是，体外细胞摄取的共聚焦激光扫描显微镜检查显示，AraHH001被mTECs同化，并在酸性隔室中共定位，如用Lysotracker Red标记所检测到的。因此，本文首次报道的与mTECs结合的特异性DNA适体的开发，不仅作为治疗适体，而且作为似乎在血管生成中起主要作用的靶向分子探针，以及用于靶向新药物递送系统的开发，都具有很大的前景。
The present study used a spontaneous cell-based SELEX method (Systemic Evolution of Ligands by EXponential Enrichment) to produce DNA aptamers that specifically bind to cell surface proteins or biomarkers produced by primary cultured mouse tumor endothelial cells (mTECs). In solid tumors, new blood vessels are formed through an angiogenesis process, and this plays a critical role in cancer development as well as metastasis. To combat angiogenesis, an appropriate diagnosis and a molecular-level understanding of the different cancer types are now a high priority. The novel DNA aptamer AraHH001, developed in this study, binds specifically to mTECs with high affinity in the nano-molar range, but does not bind to normal skin endothelial cells (skin-ECs). The selected DNA aptamer was also found to bind to cultured human tumor endothelial cells (hTECs), isolated from a clinical patient with a renal carcinoma. The aptamer AraHH001 showed significant anti-angiogenesis activity by inhibiting tube formation by mTECs on matrigel. Interestingly, a confocal laser scanning microscopy examination of in vitro cellular uptake revealed that AraHH001 was assimilated by mTECs, and became co-localized in acidic compartments, as detected by labeling with Lysotracker Red. Therefore, the development of a specific DNA aptamer that binds to mTECs, as reported here for the first time, holds great promise not only as a therapeutic aptamer but also as a targeted molecular probe that appears to play a major role in angiogenesis, and for the development of a targeted new drug delivery system.
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DOI：
10.1073/pnas.85.20.7652
发表时间：
1988-10-01
影响因子：
11.1
作者：
GYLLENSTEN, UB;ERLICH, HA
通讯作者：
ERLICH, HA
影响因子：
14.9
作者：
Ferreira CS;Cheung MC;Missailidis S;Bisland S;Gariépy J
通讯作者：
Gariépy J
影响因子：
8.8
作者：
Kurosu, T.;Ohga, N.;Hida, Y.;Maishi, N.;Akiyama, K.;Kakuguchi, W.;Kuroshima, T.;Kondo, M.;Akino, T.;Totsuka, Y.;Shindoh, M.;Higashino, F.;Hida, K.
通讯作者：
Hida, K.
影响因子：
7
作者：
Mallikaratchy, Prabodhika;Tang, Zhiwen;Tan, Weihong
通讯作者：
Tan, Weihong
影响因子：
3.7
作者：
Kandimalla KK;Scott OG;Fulzele S;Davidson MW;Poduslo JF
通讯作者：
Poduslo JF