Sensitive detection and quantitation of EZH2 expression in cancer cell by an electrochemiluminescent method

Qiang Li; Xiaoming Zhou

doi:10.1117/12.843394

28 October 2009 Sensitive detection and quantitation of EZH2 expression in cancer cell by an electrochemiluminescent method

Qiang Li, Xiaoming Zhou

Author Affiliations +

Proceedings Volume 7519, Eighth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2009); 75191J (2009) https://doi.org/10.1117/12.843394
Event: Photonics and Optoelectronics Meetings 2009, 2009, Wuhan, China

Abstract

The polycomb group protein enhancer of zeste homolog 2 (EZH2) regulating cell cycle and functioning as a transcriptional repressor, is overexpressed in several human cancers. Therefore it can be a molecular marker for detection of cancer progression and metastasis. Here the electrochemiluminescence (ECL) assay was developed to detect and quantify the amount of EZH2 mRNA expression in cancer cell. Total mRNA was reverse transcribed into cDNA. The cDNA was amplified using a forward and reverse primer pair which were labeled with biotin and Tris (2, 2-bipyridine) ruthenium (II) (TBR) on the 5' end, respectively. The amplification product was captured on streptavidin coated magnetic beads and then separated using a magnetic field. The TBR labels were reacted with the most efficient coreactant, TPA, on the electrode. Photons were produced and detected by a custom-built ECL system. The housekeeping gene hydroxymethylbilane synthase (HMBS) was used as an approximate reference to quantify the amount of EZH2 mRNA expression, whose primer pairs were labeled the same as EZH2. Result indicated that the EZH2 mRNA was overexpressed in MCF-7 cells relative to normal blood cells. This assay is specific and sensitive and could be used for the clinical diagnosis and prognosis of cancer.

Citation Download Citation

Qiang Li and Xiaoming Zhou "Sensitive detection and quantitation of EZH2 expression in cancer cell by an electrochemiluminescent method", Proc. SPIE 7519, Eighth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2009), 75191J (28 October 2009); https://doi.org/10.1117/12.843394

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