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用于检测雌激素受体 α 阳性癌的 DNA 适体探针。

DNA aptamer probes for detection of estrogen receptor α positive carcinomas.

机构信息

Bioengineering Research Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, India.

Department of Surgical Oncology, Dr. Bhubaneswar Borooah Cancer Institute, Guwahati, Assam, India; Mugagen Laboratories Pvt. Ltd, Technology Incubation Centre, Indian Institute of Technology Guwahati, Guwahati, Assam, India.

出版信息

Transl Res. 2017 May;183:104-120.e2. doi: 10.1016/j.trsl.2016.12.008. Epub 2016 Dec 25.

Abstract

Estrogen receptor alpha (ERα) also known as NR3A1 (nuclear receptor subfamily 3, group A, member 1) is a ligand-activated transcription factor. It is an important biomarker for breast cancer metastasis. In the present study, we report a novel DNA aptamer candidate against estrogen receptor (ER) alpha structure. The enriched aptamer candidate was obtained after 14 iterative cycles of in vitro protein-SELEX process. Isothermal calorimetry study suggests the nanomolar sensitivity of the candidate ER_Apt1 to its target protein. Fluorescence- and chemiluminescence-binding assays confirm the specificity of the candidate aptamer to ER alpha positive breast cancer cell line. Comparative analysis of ER_Apt1 to ER alpha monoclonal antibody was also performed to analyze the expression of ER alpha in various malignant cancer cell line. Cytochemical and immunohistochemistry assay indicates its potential use as a diagnostic agent against ERα positive carcinomas. The nucleotide aptamer sequences described in the present study can be used for the detection, treatment, prophylaxis and diagnosis of ERα-related disorder.

摘要

雌激素受体 alpha(ERα)也被称为 NR3A1(核受体亚家族 3,A 组,成员 1),是一种配体激活的转录因子。它是乳腺癌转移的重要生物标志物。在本研究中,我们报告了一种针对雌激素受体(ER)alpha 结构的新型 DNA 适体候选物。经过 14 轮体外蛋白 SELEX 过程的富集,得到了富含适体的候选物。等温量热法研究表明,候选物 ER_Apt1 对其靶蛋白的纳米摩尔敏感性。荧光和化学发光结合分析证实了候选适体对 ERα阳性乳腺癌细胞系的特异性。还对 ER_Apt1 与 ERα单克隆抗体进行了比较分析,以分析 ERα在各种恶性癌细胞系中的表达。细胞化学和免疫组织化学分析表明,它有潜力作为 ERα阳性癌的诊断试剂。本研究中描述的核苷酸适体序列可用于检测、治疗、预防和诊断与 ERα相关的疾病。

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