title: Label-free electrochemical detection of human methyltransferase from tumors.
abstract: The role of abnormal DNA methyltransferase activity in the development and progression of cancer is an essential and rapidly growing area of research, both for improved diagnosis and treatment. However, current technologies for the assessment of methyltransferase activity, particularly from crude tumor samples, limit this work because they rely on radioactivity or fluorescence and require bulky instrumentation. Here, we report an electrochemical platform that overcomes these limitations for the label-free detection of human DNA(cytosine-5)-methyltransferase1 (DNMT1) methyltransferase activity, enabling measurements from crude cultured colorectal cancer cell lysates (HCT116) and biopsied tumor tissues. Our multiplexed detection system involving patterning and detection from a secondary electrode array combines low-density DNA monolayer patterning and electrocatalytically amplified DNA charge transport chemistry to measure selectively and sensitively DNMT1 activity within these complex and congested cellular samples. Based on differences in DNMT1 activity measured with this assay, we distinguish colorectal tumor tissue from healthy adjacent tissue, illustrating the effectiveness of this two-electrode platform for clinical applications.
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and.
authors.forename: Ariel L
authors.initials: AL
authors.lastname: Furst
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and.
authors.forename: Natalie B
authors.initials: NB
authors.lastname: Muren
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and Department of Chemistry and Chemical Biology, Occidental College, Los Angeles, CA 90041.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Hill
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125; and jkbarton@caltech.edu.
authors.forename: Jacqueline K
authors.initials: JK
authors.lastname: Barton
mesh_terms: D002384:Catalysis; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D019175:DNA Methylation; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D045325:HCT116 Cells; D006801:Humans; D009369:Neoplasms / Q000378:metabolism*
chemical_list: D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human
keywords: DNA electrochemistry; electrocatalysis; methylation detection