Liu Wenjie, Mohan Sruthi Priya, Nagaraj Nareshkumar Ragavachetty, Sundar Jaganathan Shyam, Wen Yi, Ramasubramanyan Sharada, Irudayaraj Joseph
Department of Bioengineering, Cancer Center at Illinois, Micro and Nanotechnology Laboratory. University of Illinois at Urbana-Champaign, Urbana, IL.
Biomedical Research Center in Mills Breast Cancer Institute, Carles Foundation Hospital, Urbana, IL.
Mol Vis. 2021 Nov 20;27:643-655. eCollection 2021.
To elucidate the mechanism behind epigenetic alteration associated with dexamethasone (DEX) sodium phosphate treatment.
We performed enzyme-linked immunosorbent assay to quantify changes in global DNA methylation and hydroxymethylation, quantitative real-time PCR (qRT-PCR) of the DNA methylation- and hydroxymethylation-related gene, in vitro DNA methyltransferase (DNMT) enzymatic activity assays with purified DNMTs, and DNA hydroxymethylation pattern with super-resolution imaging.
We identified global DNA hypomethylation and hyper-hydroxymethylation upon DEX treatment, associated with aberrant mRNA expression levels of DNMT and ten-eleven translocation (TET) proteins. Additionally, DEX exposure could directly hinder DNMT activities.
We showed that DEX-induced epigenetic alterations are linked to aberrant DNMT and TET expression, potentially through an essential role of DNMT.
阐明与地塞米松磷酸钠治疗相关的表观遗传改变背后的机制。
我们进行了酶联免疫吸附测定以量化整体DNA甲基化和羟甲基化的变化，对DNA甲基化和羟甲基化相关基因进行定量实时PCR（qRT-PCR），用纯化的DNA甲基转移酶（DNMT）进行体外DNA甲基转移酶活性测定，并用超分辨率成像分析DNA羟甲基化模式。
我们发现地塞米松治疗后整体DNA低甲基化和高羟甲基化，这与DNMT和十一-易位（TET）蛋白的异常mRNA表达水平相关。此外，地塞米松暴露可直接阻碍DNMT活性。
我们表明地塞米松诱导的表观遗传改变与DNMT和TET的异常表达有关，可能是通过DNMT的重要作用。
