Construction Of A Functional Detection System For Drug Induced Liver Injury Associated DNA Methylation

Drug-induced liver injury is a life-threatening drug adverse reaction.Drug-induced liver injury appears and acts in a distinctive way in different patients who use exactly identical drug.Currently,there are more than 1100 types of common clinical drugs globally that have potential hepatotoxicity,including anti-tuberculosis,anti-tumors,herbals and so on.These drugs are extremely necessary in curing their target diseases,and it is difficult to avoid liver injury by replacing these drugs directly.Studies have shown that the subtle difference of DNA methylation is one of the explanations of such individual difference in drug-induced liver injury.Although association study suggested the correlation between drug-induced liver injury and hypermethylation in promoter region of gene CYP2D6 and CYP2E1,however,there is little experiment methods to verify this connection between drug-induced liver injury and DNA methylation.Therefore,a druginduced liver injury predictable methylation functional detection human hepatic system is urgently needed.In this study,we built a novel methylation functional detection hepatic cell model,choosed the detection assay related to drug-induced liver injury(cell viability,extracellular lactose dehydrogenase content,intracellular glutathione content and intracellular Caspase3/7 activity)on the basis of this model and established a methylation functional detection system associated to drug-induced liver injury.Our results showed the correlation between drug-induced liver injury and hypermethylation in promoter region of gene CYP2D6 and CYP2E1.The results demonstrated that the methylation level of gene CYP2D6 and CYP2E1 was inversely proportional to the cell viability and intracellular glutathione,while it was proportional to the intracellular Caspase3/7 activity in HepaRG cells,after cells were treated with rifampin,a kind of antituberculosis.It was the first time that the correlation between hypermethylation in promoter region of gene CYP2D6 and CYP2E1 and drug-induced liver injury was tested in vitro.We expect this well-designed methylation detection model will serve as a useful tool in validating correlation between DNA methylation and drug-induced liver injury in in vitro system.This study can also provide clues for future investigations to unveil more details of the underlying mechanisms of drug-induced liver injury and facilitate in advancing personalized medicine design.