Zhu Sha, Cao Ya, Xu Yuanyuan, Yin Yongmei, Li Genxi
Department of Oncology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
Int J Mol Sci. 2013 May 16;14(5):10298-306. doi: 10.3390/ijms140510298.
Estrogen receptor (ER), expressed in approximately 80% of primary breast cancer cells, has proven to be a valuable predictive factor of the disease. Herein, by making use of the specific binding of ER to its DNA response elements, we propose an Exonuclease III (Exo III) protection-based electrochemical method for detecting ER proteins. In this assay, the presence of ER can protect the duplex DNA molecules immobilized on an electrode surface from Exo III-catalyzed digestion, resulting in an increased electrochemical signal. Experimental results have revealed that the proposed method can allow the quantification of ER in the range of 0.5 to 100 nM with a satisfactory detection limit of 0.38 nM. Furthermore, since this approach can also be employed to detect ER directly in nuclear extracts, it may be of great use in biomedical applications in the future.
雌激素受体(ER)在大约80%的原发性乳腺癌细胞中表达,已被证明是该疾病的一个有价值的预测因子。在此,通过利用ER与其DNA反应元件的特异性结合,我们提出了一种基于核酸外切酶III(Exo III)保护的电化学方法来检测ER蛋白。在该检测中,ER的存在可以保护固定在电极表面的双链DNA分子免受Exo III催化的消化,从而导致电化学信号增强。实验结果表明,所提出的方法能够在0.5至100 nM的范围内对ER进行定量,检测限为0.38 nM,令人满意。此外,由于该方法也可用于直接检测核提取物中的ER,未来在生物医学应用中可能会有很大用途。