An Exonuclease III Protection-Based Electrochemical Method for Estrogen Receptor Assay

Zhu, Sha; Cao, Ya; Xu, Yuanyuan; Yin, Yongmei; Li, Genxi

doi:10.3390/ijms140510298

Cited by 11 publications

(6 citation statements)

References 23 publications

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“…676 An exonuclease III protection-based voltammetric assay for ER was designed using a DNA duplex, incorporating an ERspecific sequence, immobilized onto the surface of a gold electrode. 677 The second strand was labeled with methylene blue at the proximal end. The presence of ER protected the duplex DNA molecules from exonuclease III digestion, resulting in an electrochemical signal.…”

Section: Vascular Endothelial Growth Factor (Vegf)mentioning

confidence: 99%

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Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Sargent²,

2016

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Rapid progress in identifying biomarkers that are hallmarks of disease has increased demand for high-performance detection technologies. Implementation of electrochemical methods in clinical analysis may provide an effective answer to the growing need for rapid, specific, inexpensive, and fully automated means of biomarker analysis. This Review summarizes advances from the past 5 years in the development of electrochemical sensors for clinically relevant biomolecules, including small molecules, nucleic acids, and proteins. Various sensing strategies are assessed according to their potential for reaching relevant limits of sensitivity, specificity, and degrees of multiplexing. Furthermore, we address the remaining challenges and opportunities to integrate electrochemical sensing platforms into point-of-care solutions.

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Section: Vascular Endothelial Growth Factor (Vegf)mentioning

confidence: 99%

“…An exonuclease III protection-based voltammetric assay for ER was designed using a DNA duplex, incorporating an ER-specific sequence, immobilized onto the surface of a gold electrode . The second strand was labeled with methylene blue at the proximal end.…”

Section: Electrochemical Analysis Of Proteinsmentioning

confidence: 99%

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Sargent²,

2016

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“…Several experiments have been developed for the comprehension and for the characterization of the mechanism by which ER is activated by ligand and its binding capacity as a homodimer (ER/E2) 2 to a specific DNA consensus sequence (ERE) located upstream of some estrogenic regulated genes. In this way several biophysical tools have been developed to explore the interaction phenomena such as fluorescence anisotropy [6][7][8], surface plasmon resonance (SPR) [9][10][11][12], FRET [13], electrochemical [14][15][16], resonant waveguide [17]. The main objectives were to determine the estrogenic compounds/ER interaction properties [18][19][20], the ER/ER dimerization phenomena [13,21], ER/ERE interaction mechanism and properties under estrogenic stimulation [9,10].…”

Section: Introductionmentioning

confidence: 99%

Impact of biochemical design on estrogen receptor/estrogen response element interaction by surface plasmon resonance technology

Habauzit

Bayle

Bénimèlis

et al. 2014

Archives of Biochemistry and Biophysics

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The estrogen receptor (ER) is a transcription factor that binds under 17-β-estradiol (E2) stimulation as homodimer to a short DNA consensus sequence named estrogen response element (ERE). The ER/ERE interaction has been assessed by several research groups through different methodologies notably by surface plasmon resonance (SPR) techniques. The biochemical parameters and conditions (solvent, ER concentration, salt, time and temperature) used to prepare samples before analysis were very different from one study to another. But no studies have aimed to compare the effect of these modifications on ER/ERE interaction. Therefore the main objective of the present paper was to assess the influence of biochemical parameters onto the ER/ERE interaction with the final aim to improve the comprehension of this interaction. Our results highlighted that parameters like solvent, ER concentration, salt and surfactant concentration, temperature and time deeply modify ER/ERE interaction. Nevertheless, the dimer formation under E2 stimulation occurred with all tested conditions. Altogether, incubation parameters of ER with E2, deeply modify its binding level onto ERE. These data constitute an important key point to consider for the improvement of ER/ERE detection method depending upon the aim of the study (interaction measurement, environmental detection, development of new technologies or devices).

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“…Recently, we have reported that ER binding to the dsDNA molecules that contain the consensus estrogen response elements (EREs) could protect the dsDNA from Exo IIImediated digestion, which formed a basis for constructing an electrochemical biosensor to detect ER proteins [4]. In the present work, we apply the assay method to further investigate the effect of PI3K inhibitor on the ER protein expression.…”

Section: Resultsmentioning

confidence: 94%

Electrochemical assay of the relationship between the inhibition of phosphatidylinositol 3-kinase pathway and estrogen receptor expression in breast cancer

Zhu

Cao

et al. 2013

Anal Bioanal Chem

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Breast cancer has become one of the most threatening diseases to women throughout the world. Emerging evidence implies that estrogen receptor (ER) and phosphatidylinositol 3-kinase (PI3K) pathways play central roles in both breast cancer progression and response to therapy. In this work, we have probed into ER expression related to the PI3K pathway at the protein level with an electrochemical technique based on the detection of ER proteins in nuclear extracts with an Exonuclease III protection-based strategy. Experimental results show that an increased number of ER proteins can be detected upon PI3K inhibition, demonstrating the reversal effect of the PI3K inhibitor on ER expression. Moreover, treatment with different concentrations of the PI3K inhibitor NVP-BEZ235 can result in a dose-dependent alteration of ER protein levels, implying an intimate link between ER and PI3K pathways. This work may be a great help to understand the mysteries underlying PI3K-related endocrine resistance and to evaluate the effect of therapeutic interventions in the future.

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An Exonuclease III Protection-Based Electrochemical Method for Estrogen Receptor Assay

Cited by 11 publications

References 23 publications

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules

Impact of biochemical design on estrogen receptor/estrogen response element interaction by surface plasmon resonance technology

Electrochemical assay of the relationship between the inhibition of phosphatidylinositol 3-kinase pathway and estrogen receptor expression in breast cancer

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