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    Genosensor made with a self-assembled monolayer matrix to detect MGMT gene methylation in head and neck cancer cell lines

    Year: 2020

    Journal: Talanta, Volume 210, APR 1

    Authors: Carr, O; Raymundo-Pereira, PA; Shimizu, FM; Sorroche, BP; Melendez, ME; Pedro, RD; Miranda, PB; Carvalho, AL; Reis, RM; Arantes, LMRB; Oliveira, ON

    Organizations: CNPqConselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPQ) [423952/2018-8]; INEO; FAPESPFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2019/13390-8, 2015/12990-0, 2012/15543-7, 2018/02819-0, 2017/03503-4, 2013/14262-7, 2016/01919-6]

    Keywords: DNA methylation; Head and neck cancer; Genosensor; Impedance spectroscopy; MGMT gene; Biosensor

    DNA methylation is involved in the oncogenesis of head and neck squamous cell carcinoma and could be used for early detection of cancer to increase the chances of cure, but unfortunately diagnosis is usually made at late stages of the disease. In this work we developed genosensors to detect DNA methylation of the MGMT gene in head and neck cancer cell lines. The probe for MGMT promoter methylation was immobilized on gold electrodes modified with 11-mercaptoundecanoic acid (11-MUA) self-assembled monolayers (SAM). Detection was performed with electrochemical impedance spectroscopy, with clear distinction between methylated and non-methylated DNA from head and neck cell lines. The genosensor is sensitive with a low detection limit of 0.24 x 10(-12) mol L-1. In addition, the cell lines FaDu, JHU28 and SCC25 for the MGMT gene, could be distinguished from the HN13 cell line which has a high degree of MGMT methylation (97%), thus confirming the selectivity. Samples with different percentages of MGMT DNA methylation could be separated in multidimensional projections using the visualization technique interactive document mapping (IDMAP). The genosensor matrix and the immobilization procedures are generic, and can be extended to other DNA methylation biomarkers.
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