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具有高亲和力和特异性的抗表皮生长因子受体DNA适配体的筛选。

Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity.

作者信息

Wang Deng-Liang, Song Yan-Ling, Zhu Zhi, Li Xi-Lan, Zou Yuan, Yang Hai-Tao, Wang Jiang-Jie, Yao Pei-Sen, Pan Ru-Jun, Yang Chaoyong James, Kang De-Zhi

机构信息

The First Clinical Medical College of Fujian Medical University, Fuzhou, China; Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.

State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.

出版信息

Biochem Biophys Res Commun. 2014 Oct 31;453(4):681-5. doi: 10.1016/j.bbrc.2014.09.023. Epub 2014 Sep 19.

Abstract

Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher's attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56±7.3nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

摘要

表皮生长因子受体(EGFR/HER1/c-ErbB1)在许多实体癌中过表达,如表皮样癌、恶性胶质瘤等。EGFR在恶性癌细胞的增殖、侵袭、血管生成和转移中发挥作用,是癌症检测、成像和治疗临床应用的理想抗原。适体是通过指数富集配体系统进化(SELEX)产生的DNA/RNA寡核苷酸,能够特异性结合蛋白质和其他物质。RNA适体由于其不稳定性和高生产成本而不适用。相反,DNA适体因其易于合成、稳定、具有选择性、具有高结合亲和力且生产成本效益高而引起了研究人员的关注。在本研究中,我们成功鉴定出对EGFR具有高结合亲和力和选择性的DNA适体。从鉴定出的DNA适体中选择了Kd为56±7.3nM的名为TuTu22的适体进行进一步研究。流式细胞术分析结果表明,TuTu22适体能够特异性识别多种表达EGFR的癌细胞,但不与EGFR阴性细胞结合。鉴于上述所有优点,本文报道的针对癌症生物标志物EGFR的DNA适体将有助于新型靶向癌症检测、成像和治疗的发展。

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