Effects of PM2.5 on Human Placental Trophoblast Cells by iTRAQ Combined with 2D LC-MS/MS

QIN Zhe; FU Feng; XU Zhong-wei; HOU Hai-yan; CHEN Ya-qiong

doi:10.3969/j.issn.1674-1870.2017.02.020

Abstract：Objective:To preliminarily screened the protein molecules and biological processes that may be involved in PM2.5 increases the rate of adverse pregnancy by affecting the placental trophoblast cells using proteomic technology.Methods:HTR-8/SVneo cells were exposed to 0,30,60,120 and 200 μg/mL PM2.5 for 24 h and 48 h,respectively,to observe the cell proliferation;finally,we selected 120 μg/mL PM2.5 treated with HTR-8/SVneo cells for 24 h and 48 h for follow-up experiments.After extraction of intracellular protein and enzyme digestion,peptides and differentially expressed proteins were analyzed and identified by isobaric tags for relative and absolute quantitation (iTRAQ) combined with two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) technology.Finally,bioinformatics analysis of the data was carried out.Results:The survival rates of cells treated with 120 μg/mL PM2.5 for 24 h and 48 h were 86.09％ and 52.36％,respectively,which were also remarkably decreased compared with that of control cells (P＜0.05).There were 182 differentially expressed proteins in the cells exposure to 120 μg/mL PM2.5 for 24 h,involved in 22 cell biological processes (BP);meanwhile,cells exposed for 48 h had 486 differential proteins,involving 217 BP.Conclusions:PM2.5 can affect a large number of BP in HTR-8/SVneo cells.iTRAQ with LC-MS/MS 2D technology is an effective way for high sensitivity and throughput screening of differentially expressed proteins in cells after exposure to PM2.5,which provides a basis for the future research on the mechanism of PM2.5 increased the risk of adverse pregnancy outcomes.