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Analysis of toxicity effects of Di-(2-ethylhexyl) phthalate exposure on human bronchial epithelial 16HBE cells

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Abstract

Recent studies have indicated that Di-(2-ethylhexyl) phthalate (DEHP), the most commonly used plasticizer in daily-life products, could be dispersed in indoor air and induce human exposure via inhalation. DEHP has been reported to have effects on the respiratory system in both animal and human researches. The toxicity effects of DEHP exposure on cell proliferation, cell cycle progression, apoptosis, global DNA methylation and the expression levels of DNA methyltransferases (DNMTs) were investigated in this study, using human epithelial cell line 16HBE as an in vitro model. Cells were treated with DEHP at doses of 0, 0.125, 0.5 and 2 mmol/L for 48 h. Cell proliferation, cell cycle and apoptosis were tested by MTT assay and flow cytometer, respectively. The obtained results showed decreased living cell number and cell viability following DEHP exposure at the dose of 2 mmol/L. DEHP also inhibited the cell cycle progression of G1 phase and induced a significant increase in cell apoptosis in 16HBE cells. DEHP exposure could induce cell proliferation inhibition in 16HBE cells via the blocking of cell cycle progression and accelerated cell apoptosis. In addition, decreased global DNA methylation levels and expression levels of DNMTs were observed in DEHP-treated groups which revealed possible epigenetic effects of DEHP.

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Abbreviations

DEHP:

Di-(2-ethylhexyl) phthalate

DMSO:

Dimethyl sulfoxide

DNMT1:

DNA methyltransferase 1

FBS:

Fetal bovine serum

MEHP:

Mono-2-ethylhexyl phthalate

MEM:

Minimum essential medium

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

OD:

Optical density

PBS:

Phosphate buffer saline

PI:

Prodidium iodide

PVC:

Polyvinyl chloride

QPCR:

Quantitative real-time PCR

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Acknowledgements

This work was supported by funds from the National Natural Science Foundation of China (81072323, 81602831), China Postdoctoral Science Foundation funded project (2016M602537) and the Guangdong Provincial Science and Technology Project (2013B021800032). We offer our sincere thanks to all of the participants.

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  1. Yue Ma and Yinsheng Guo have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Molecular Biology, Shenzhen Center for Disease Control and Prevention, Shenzhen, 518055, People’s Republic of China

    Yue Ma, Yinsheng Guo, Shuang Wu, Ziquan Lv, Qian Zhang, Xing Xie & Yuebin Ke

  2. Key Laboratory of Environment and Health (HUST), Ministry of Education & Ministry of Environmental Protection, and State Key Laboratory of Environmental Health (Incubation), School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, People’s Republic of China

    Yue Ma

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  1. Yue Ma
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Correspondence to Yuebin Ke.

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Ma, Y., Guo, Y., Wu, S. et al. Analysis of toxicity effects of Di-(2-ethylhexyl) phthalate exposure on human bronchial epithelial 16HBE cells. Cytotechnology 70, 119–128 (2018). https://doi.org/10.1007/s10616-017-0111-6

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  • DOI: https://doi.org/10.1007/s10616-017-0111-6

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