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Identification of an aptamer targeting hnRNP A1 by tissue slide-based SELEX
Li, Shaohua1; Xu, Hua1; Ding, Hongmei1; Huang, Yanping2; Cao, Xiaoxiao1; Yang, Guang1; Li, Jie1; Xie, Zhigang3; Meng, Yuhong4; Li, Xiaobing5; Zhao, Qiang1; Shen, Beifen1; Shao, Ningsheng1
关键词cancerous tissue slide SELEX breast cancer aptamer heterogeneous nuclear ribonucleoprotein A1 biomarker
刊名JOURNAL OF PATHOLOGY
2009-07-01
DOI10.1002/path.2543
218期:3页:327-336
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Oncology ; Pathology
研究领域[WOS]Oncology ; Pathology
关键词[WOS]HETEROGENEOUS NUCLEAR RIBONUCLEOPROTEINS ; CANCER-CELLS ; CONJUGATED NANOPARTICLES ; EXPONENTIAL ENRICHMENT ; SYSTEMATIC EVOLUTION ; TELOMERIC REPEATS ; LUNG-CANCER ; LIVE CELLS ; IN-VITRO ; LIGANDS
英文摘要

We report a new in situ tissue slide-based SELEX strategy targeting neoplastic tissues from breast cancer patients. The methodology, using the molecular differences between clinical specimens, can evolve aptamers to all fractions of tissue. The aptamers may be used as new molecular probes for pathological diagnosis and tumour imaging, and also to reveal the molecular differences that are responsible for the diseases. The specific aptamers were enriched by unequal length strand PCR employing a structured (-) strand primer. After 12 rounds of selection, using the paraffin tissue sections from infiltrating ductal carcinomas as targets, and using the adjacent normal tissue from the same case as controls, one of the enriched ssDNA aptamers, BC15, was selected from a nucleic acid library and characterized as recognizing breast cancer cells either within the tissue sections or from the culture medium, but only weakly binding to adjacent normal cells or immortalized breast cell line MCF10A. The calculated equilibrium dissociation constants (K(d)) of BC15 bound to MCF7 cells was 111.0 +/- 36.9 nM. Through streptavidin magnetic beads mediated affinity purification assay followed by mass spectrometry identification and western blot confirmation, the target of BC15 was characterized to be hnRNP A1, which was further verified to be specifically and highly expressed in cancerous tissues of breast by hnRNP A1 antibody immunostaining as well as western blot. BC15 aptamer was also used to probe cancer cells in tissues from other pathological types of breast cancers including lobular carcinoma, ductal carcinoma complicated with lobular carcinoma, comedo carcinoma, and lymph node metastasis of breast ductal carcinoma origin or breast lobular carcinoma origin. Therefore, tissue slide-based SELEX holds promise in identifying tumour markers and developing specific molecular probes for cancer diagnosis. Copyright (C) 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

语种英语
WOS记录号WOS:000267613300005
项目编号2006AA02Z408 ; 2005CB724600
资助机构Chinese National High-Tech Research and Development Programme ; Chinese National Key Programme on Basic Research
引用统计
被引频次:77[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/61725
专题北京大学第三临床医学院_病理科
作者单位1.Beijing 307 Hosp, Dept Pathol, Beijing, Peoples R China
2.Naval Gen Hosp, Dept Pathol, Beijing, Peoples R China
3.Beijing Inst basic Med Sci, Dept Biochem & Mol Biol, Beijing, Peoples R China
4.Chinese Peoples Armed Police Forces, Gen Hosp, Dept Gastroenterol, Beijing, Peoples R China
5.Peking Univ, Hosp 3, Dept Pathol, Beijing 100871, Peoples R China
推荐引用方式
GB/T 7714
Li, Shaohua,Xu, Hua,Ding, Hongmei,et al. Identification of an aptamer targeting hnRNP A1 by tissue slide-based SELEX[J]. JOURNAL OF PATHOLOGY,2009,218(3):327-336.
APA Li, Shaohua.,Xu, Hua.,Ding, Hongmei.,Huang, Yanping.,Cao, Xiaoxiao.,...&Shao, Ningsheng.(2009).Identification of an aptamer targeting hnRNP A1 by tissue slide-based SELEX.JOURNAL OF PATHOLOGY,218(3),327-336.
MLA Li, Shaohua,et al."Identification of an aptamer targeting hnRNP A1 by tissue slide-based SELEX".JOURNAL OF PATHOLOGY 218.3(2009):327-336.
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