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学科主题: 临床医学
题名:
Identification of an aptamer targeting hnRNP A1 by tissue slide-based SELEX
作者: 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
发表日期: 2009-07-01
DOI: 10.1002/path.2543
卷: 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
所属项目编号: 2006AA02Z408 ; 2005CB724600
项目资助者: Chinese National High-Tech Research and Development Programme ; Chinese National Key Programme on Basic Research
WOS记录号: WOS:000267613300005
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/61725
Appears in Collections:北京大学第三临床医学院_病理科_期刊论文

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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

Recommended Citation:
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.
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