Perfluorobutane sulfonate exposure disrupted human placental cytotrophoblast cell proliferation and invasion involving in dysregulating preeclampsia related genes

Abstract

We reported that maternal PFBS, an emerging pollutant, exposure is positively associated with preeclampsia which can result from aberrant trophoblasts invasion and subsequent placental ischemia. In this study, we investigated the effects of PFBS on trophoblasts proliferation/invasion and signaling pathways. We exposed a human trophoblast line, HTR8/SVneo, to PFBS. Cell viability, proliferation, and cell cycle were evaluated by the MTS assay, Ki-67 staining, and flow cytometry, respectively. We assessed cell migration and invasion with live-cell imaging-based migration assay and matrigel invasion assay, respectively. Signaling pathways were examined by Western blot, RNA-seq, and qPCR. PFBS exposure interrupted cell proliferation and invasion in a dose-dependent manner. PFBS (100 μM) did not cause cell death but instead significant cell proliferation without cell cycle disruption. PFBS (10 and 100 μM) decreased cell migration and invasion, while PFBS (0.1 μM) significantly increased cell invasion but not migration. Further, RNA-seq analysis identified dysregulated HIF-1α target genes that are relevant to cell proliferation/invasion and preeclampsia, while Western Blot data showed the activation of HIF-1α, but not Notch, ERK1/2, (PI3K)AKT, and P38 pathways. PBFS exposure altered trophoblast cell proliferation/invasion which might be mediated by preeclampsia-related genes, suggesting a possible association between prenatal PFBS exposure and adverse placentation. K E Y W O R D S invasion, PFBS, placenta cytotrophoblast, preeclampsia, proliferation | 14183 MARINELLO Et AL. 1 | INTRODUCTION Poly-and per-fluoroalkyl substances (PFAS) have attracted widespread attention in recent years due to their bioaccumulation, toxicity, and ubiquitous nature. 1 PFAS are a group of compounds characterized by a hydrophobic poly-fluorinated alkyl chain and a polar hydrophilic terminal functional group. PFAS are used in a variety of industrial and consumer products such as surfactants for soil/stain resistance, textiles, paper and metals, firefighting foam, and pesticides. 2,3 Humans are exposed to PFAS through contaminated drinking water, food, outdoor air, indoor dust, and soil. 4 One of the most widely known PFAS is perfluorooctane sulfonic acid (PFOS), which has an eight-carbon backbone with a sulfonate. Due to strong carbon-fluorine bonds (C8), PFOS is extremely stable and persistent in the environment (USEPA, Document# 822R14002) and is not readily eliminated from humans due to its half-life of 5.4 years. 5-16 Data from human and animal studies demonstrate numerous health and ecological risks resulting from PFOS exposure including increased risk of thyroid disease, blood cholesterol levels, and preeclampsia and decreased body's response to vaccine, fertility in women, and birth weight, liver, and immune system damage. 17-29 Thus, beginning in 2002, most manufacturing of PFOS in the United States was discontinued voluntarily by 3M and DuPont in favor of shorter chain PFAS (C4 or C6, Toni Krasnic, the U. S. Environmental Protection A...

A High-Throughput Toxicity Screen of 42 Per- and Polyfluoroalkyl Substances (PFAS) and Functional Assessment of Migration and Gene Expression in Human Placental Trophoblast Cells

“…Failure of trophoblast migration and invasion in vascular remodeling are associated with preeclampsia and intrauterine fetal growth restriction ( Khong et al, 1986 ). Our data suggest the potential for certain PFAS to disrupt the normal migration and invasion processes of placental trophoblasts and is consistent with inhibition of trophoblast migration and invasion by PFBS, PFOA, PFOS, and GenX in HTR-8/SVneo cells ( Marinello et al 2020 Szilagyi et al, 2020 ; Li et al, 2021 ). Furthermore, we observed gene expression changes in GenX-exposed JEG-3 cells that are consistent with inhibition of trophoblast migration, such as reduced expression of IGF2 ( Irving and Lala, 1995 ), reduced expression of EGFR ( Massimiani et al, 2015 ), and increased expression of CTNNB1 ( Tucker and Nowak, 2018 ) .…”

Perfluoroalkyl Substance Exposure and the BDNF Pathway in the Placental Trophoblast

“…The following methods for immunofluorescence staining and Western blot were performed as described in Marinello et al. ( 62 ).…”

“…3 To ensure the effects on cellular functions are not due to cytotoxicity, we used doses at least 10 times lower than LD10 (61). The following methods for immunofluorescence staining and Western blot were performed as described in Marinello et al (62).…”

Perfluoroalkyl Mixture Exposure in Relation to Fetal Growth: Potential Roles of Maternal Characteristics and Associations with Birth Outcomes

“…Firstly, fetal reproductive hormones, such as estradiol, total testosterone, and progesterone, could be affected by in utero exposure to PFOS and PFOA, these substances could adversely affect fetal development [66]. Furthermore, PFASs exposure obstructed trophoblast cell proliferation, indicating a possible association between prenatal PFASs exposure and adverse placentation [67]. With regard to Apgar scores (activity, pulse, grimace, appearance, and respiration), in the fully adjusted models, per logarithmic unit increase in PFHpA and PFHpS exposure was associated with increased Apgar-1 scores of 0.065 (95% CI: 0.002, 0.129, p = 0.044) and 0.117 (95% CI: 0.006, 0.228, p = 0.039).…”