title: DNA-based hybridization chain reaction amplification for assaying the effect of environmental phenolic hormone on DNA methyltransferase activity.
abstract: In this work, a novel electrochemical protocol with signal amplification for determination of DNA methylation and methyltransferase activity using DNA-based hybridization chain reaction (HCR) was proposed. After the gold electrode was modified with dsDNA, it was treated with M.SssI MTase, HpaII endonuclease, respectively. And then the HCR was initiated by the target DNA and two hairpin helper DNAs, which lead to the formation of extended dsDNA polymers on the electrode surface. The signal was amplified by the labeled biotin on the hairpin probes. As a result, the streptavidin-alkaline phosphatase (S-ALP) conjugated on the electrode surface through the specific interaction between biotin and S-ALP. ALP could convert 1-naphthyl phosphate into 1-naphthol and the latter could be electrochemically oxidized, which was used to monitor the methylation event and MTase activity. The HCR assay presents good electrochemical responses for the determination of M.SssI MTase at a concentration as low as 0.0067 uni tmL(-1). Moreover, the effects of anti-cancer drug and environmental phenolic hormone on M.SssI MTase activity were also investigated. The results indicated that 5-fluorouracil and daunorubicin hydrochloride could inhibit the activity, and the opposite results were obtained with bisphenol A and nonylphenol. Therefore, this method can not only provide a platform to screen the inhibitors of DNA MTase and develop new anticancer drugs, but also offer a novel technique to investigate the possible carcinogenesis mechanism.
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, 61 Daizong street, Taian, Shandong 271018, PR China.
authors.forename: Zhenning
authors.initials: Z
authors.lastname: Xu
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, 61 Daizong street, Taian, Shandong 271018, PR China.
authors.forename: Huanshun
authors.initials: H
authors.lastname: Yin
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, 61 Daizong street, Taian, Shandong 271018, PR China.
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Han
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, 61 Daizong street, Taian, Shandong 271018, PR China. Electronic address: zhyl@sdau.edu.cn.
authors.forename: Yunlei
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Chemistry and Material Science, Shandong Agricultural University, 61 Daizong street, Taian, Shandong 271018, PR China. Electronic address: ashy@sdau.edu.cn.
authors.forename: Shiyun
authors.initials: S
authors.lastname: Ai
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D001559:Benzhydryl Compounds / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003630:Daunorubicin / Q000737:chemistry; D055664:Electrochemical Techniques; D004566:Electrodes; D052244:Endocrine Disruptors / Q000378:metabolism*; D004785:Environmental Pollutants / Q000737:chemistry / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005472:Fluorouracil / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D009284:Naphthols / Q000737:chemistry / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D010636:Phenols / Q000737:chemistry / Q000378:metabolism; D000098826:Bisphenol A Compounds
chemical_list: D000469:Alkaline Phosphatase; D001559:Benzhydryl Compounds; D004247:DNA; D015257:DNA-Cytosine Methylases; D003630:Daunorubicin; D052244:Endocrine Disruptors; D004785:Environmental Pollutants; D004800:Enzymes, Immobilized; D005472:Fluorouracil; D006046:Gold; D009284:Naphthols; D010636:Phenols; C029350:1-naphthol; C025256:nonylphenol; C081573:DNA modification methylase SssI; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: DNA methylation; Environmental phenolic hormones; Hybridization chain reaction; Inhibitors; Methyltransferase