Research On The Toxic Effect Of DEHP To Breast Cancer Cell And Identify Its Potional Biomarker For Detection

DEHP(Diethylhexyl phthalate),as one of the most widely used plasticizers in the phthalate family,can migrate and dissolve out,causing food contamination and interfering with the human endocrine system.In this paper,DEHP,the most widely used phthalate plasticizer in packaging materials,was used as the research object.Based on cell substitution toxicology technology,MCF-7 cells were selected as experimental model.The toxic mechanism of DEHP on breast cancer cells was studied from the perspective of metabolomics.At the same time,differential metabolites were screened as potential biomarkers for rapid detection DEHP residues in food packaging in the future.The cell viability of MCF-7 cells at the dosages of 10-9-10-2mol/L for DEHP 24h and 48h was studied.The cell viabilities ranged from 81.9%to 260.7%for 24h and 95.8%to 237.7%for 48h after the exposure of DEHP.According to the effect of the above dose on cell activity,metabolomics studies of MCF-7 cells exposed to 10-6mol/L DEHP were conducted.The aim was to investigate toxic effects of DEHP and identify key metabolic biomarkers of DEHP toxicity in breast cancer cell line MCF-7 by untargeted metabolomics.An untargeted analytical method was successfully established for the metabolites based on UHPLC-QTOF-MS.More than 400 metabolites in MCF-7 cells dealt with 10-6 mol/L DEHP were completely successfully detected.Based on variable VIP value>1,t-test P values<0.1,13 significant potential differential biomarkers were identified,involved amino acids and derivatives,organic acid,phospholipids and Purine nucleotides,etc.The metabolites changes indicated that DEHP can affect 9 metabolic pathways,including metabolism of riboflavin,beta alanine metabolism,nitrogen metabolism and histidine metabolism as the main four metabolic pathways.In addition,8 significant potential differential biomarkers were selected to establish their quantitative detection methods in MCF-7 cells.Quality control samples were used for PCA analysis to demonstrate the stability of the method.The established method was used to detect DEHP-contaminated cell samples to further verify the results of non-targeted screening.Three metabolites with the most significant differences among the 8 metabolites were further screened,including cytosine,gamma-L-Glutamyl-L-valine,2-hydroxyadenine and their corresponding up-down trend.These three substances could be used as the unique biomarkers of DEHP to evaluate the food-borne DEHP contamination,and this method provided a new rapid detection method for the DEHP contamination in food packaging in the future.