Abstract
Perfluorooctane sulfonate (PFOS) poses potential risks to reproduction and development. Mouse embryonic stem cells (mESCs) are ideal models for developmental toxicity testing of environmental contaminants in vitro. However, the mechanism by which PFOS affects early embryonic development is still unclear. In this study, mESCs were exposed to PFOS for 24 h, and then general cytotoxicity and pluripotency were evaluated. MTT assay showed that neither PFOS (0.2 µM, 2 µM, 20 µM, and 200 µM) nor control medium (0.1% DMSO) treatments affected cell viability. Furthermore, there were no significant differences in cell cycle and apoptosis between the PFOS treatment and control groups. However, we found that the mRNA and protein levels of pluripotency markers (Sox2, Nanog) in mESCs were significantly decreased following exposure to PFOS for 24 h, while there were no significant changes in the mRNA and protein levels of Oct4. Accordingly, the expression levels of miR-145 and miR-490-3p, which can regulate Sox2 and Nanog expressions were significantly increased. Chrm2, the host gene of miR-490-3p, was positively associated with miR-490-3p expression after PFOS exposure. Dual luciferase reporter assay suggests that miR-490-3p directly targets Nanog. These results suggest that PFOS can disturb the expression of pluripotency factors in mESCs, while miR-145 and miR-490-3p play key roles in modulating this effect.
Highlights
Perfluorooctane sulfonate (PFOS) has been widely used as a surface-active agent for a wide range of commercial, industrial and household applications, including water repellents, lubricants, paints, and fire-fighting foams [1]
Cell Culture and PFOS Treatment Mouse ES cell line D3 [American Type Culture Collection (ATCC), Manassas, VA, USA, no.CRL-11632] was kindly provided by Stem Cell Bank, Chinese Academy of Sciences. This cell line has been widely used in previous studies [37,38]. Mouse embryonic stem cells (mESCs) were grown on mouse embryonic fibroblast feeder cells (MEF) that were treated by mitomycin C in knock-out Dulbecco’s modified Eagle’s medium (Gibco BRL, Grand Island, NY) supplemented with 20% ES qualified fetal bovine serum (Gibco BRL), 0.1 mM b-mercaptoethanol (Sigma Chemical, St Louis, MO), 0.1 mM nonessential amino acids (Gibco BRL), 0.1 mM L-glutamine (Gibco BRL), 0.1 mM pyruvate sodium, 100 unit/ml penicillin/ streptomycin (Gibco BRL) and 1000 U/ml of leukemia inhibitory factor (LIF) (Millipore, Billerica, MA)
Effects of PFOS on Cell Viability, Morphology and Alkaline Phosphatase Staining in mESCs
Summary
Perfluorooctane sulfonate (PFOS) has been widely used as a surface-active agent for a wide range of commercial, industrial and household applications, including water repellents, lubricants, paints, and fire-fighting foams [1]. It has been identified in various environmental sectors, including air [2], sewage sludge [3,4], snow, lake, and surface runoff water [5]. PFOS exposure can induce neonatal death [19,20,21], delayed growth and development, and delayed eye opening in rodents [20,22,23]. Numerous studies have suggested the developmental toxicity of PFOS, little is known about the underlying molecular mechanisms
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