The development of new tools for tracking the activity of human DNA methyltransferases is an important goal given the role of this enzyme as a cancer biomarker and epigenetic modulator. However, analysis of the human DNA (cytosine-5)-methyltransferase 1 (Dnmt1) activity is challenging, especially in crude samples, because of the low activity and large size of the enzyme. Here, we report a new approach to Dnmt analysis that combines nanostructured electrodes with a digest-and-amplify strategy that directly monitors Dnmt1 activity with high sensitivity. Nanostructured electrodes are required for the function of the assay to promote the accessibility of the electrode for human Dnmt1. Moreover, DNA-templated deposition of silver nanoparticles (for signal amplification) is combined with DNA Exonuclease I digestion to yield optimal target-to-control signals. We achieve high sensitivity for the detection of human Dnmt1, and particularly Dnmt1 from crude cell lysates. Specifically, the detection limit of our electrochemical assay is 20 pM, which is 2 orders of magnitude lower than previously reported methods. In crude lysates, we detected Dnmt1 from as few as five colorectal cancer cells (HCT116). With biopsy samples, we were able to distinguish colorectal tumor tissue from healthy adjacent tissue using only 10 μg of sample. The strategy enables analysis of an important marker underlying the epigenetic basis of cancerous transformation.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1021/acssensors.8b00494 | DOI Listing |
J Agric Food Chem
January 2025
Department of Chemistry, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Inadvertent exposure to aristolochic acids (AAs) is causing chronic renal disease worldwide, with aristolochic acid I (AA-I) identified as the primary toxic agent. This study employed chemical methods to investigate the mechanisms underlying the nephrotoxicity and carcinogenicity of AA-I. Aristolochic acid II (AA-II), which has a structure similar to that of AA-I, was investigated with the same methods for comparison.
View Article and Find Full Text PDFKlin Mikrobiol Infekc Lek
March 2024
Department of Microbiology, Olomouc University Hospital, Czech Republic, e-mail:
Objectives: Staphylococcus aureus is part of the human microbiota, but at the same time, it is capable of causing a wide range of diseases. Due to the ever-increasing resistance to antimicrobial agents and the existence of methicillin-resistant S. aureus (MRSA) strains, there is a real possibility of carrying even this resistant bacterium, which can subsequently cause a severe infection.
View Article and Find Full Text PDFHum Cell
January 2025
Institute of Translational Medicine, Medical College, Yangzhou University, No. 136 Jiangyangzhonglu, Yangzhou, 225009, Jiangsu, China.
Cancer, a complicated disease characterized by aberrant cellular metabolism, has emerged as a formidable global health challenge. Since the discovery of abnormal aldolase A (ALDOA) expression in liver cancer for the first time, its overexpression has been identified in numerous cancers, including colorectal cancer (CRC), breast cancer (BC), cervical adenocarcinoma (CAC), non-small cell lung cancer (NSCLC), gastric cancer (GC), hepatocellular carcinoma (HCC), pancreatic cancer adenocarcinoma (PDAC), and clear cell renal cell carcinoma (ccRCC). Moreover, ALDOA overexpression promotes cancer cell proliferation, invasion, migration, and drug resistance, and is closely related to poor prognosis of patients with cancer.
View Article and Find Full Text PDFCell Biol Toxicol
January 2025
Department of Radiology, Shengjing Hospital of China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, 110004, Liaoning Province, China.
Thyroid cancer (THCA) is an increasingly common malignant tumor of the endocrine system, with its incidence rising steadily in recent years. For patients who experience recurrence or metastasis, treatment options are relatively limited, and the prognosis is poor. Therefore, exploring new therapeutic strategies has become particularly urgent.
View Article and Find Full Text PDFJ Gastrointest Cancer
January 2025
Division of Surgical Oncology, Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
Pancreatic ductal adenocarcinoma (PDAC) is projected to be the second leading cause of cancer-related death by 2030. Early identification is rare, with a 5-year overall survival (OS) of less than 10%. Advances in the understanding of PDAC tumor biology are needed to improve these outcomes.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!