Analysis of toxicity effects of Di-(2-ethylhexyl) phthalate exposure on human bronchial epithelial 16HBE cells
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.
KeywordsApoptosis Cell cycle DEHP DNA methylation Human bronchial epithelial 16HBE cell
DNA methyltransferase 1
Fetal bovine serum
Minimum essential medium
Phosphate buffer saline
Quantitative real-time PCR
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.
Compliance with ethical standards
Conflict of interest
The authors have no conflict of interest.
- Cadogan DF, Howick CJ (1996) Encyclopedia of chemical technology, vol. In: Othmer K (ed) Plasticizers. Wiley, New YorkGoogle Scholar
- Glue C, Millner A, Bodtger U, Jinquan T, Poulsen LK (2002) In vitro effects of monophthalates on cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells from allergic and non-allergic donors. Toxicol in vitro: an international journal published in association with BIBRA 16:657–662CrossRefGoogle Scholar
- Laing LV, Viana J, Dempster EL, Trznadel M, Trunkfield LA, Uren Webster TM, van Aerle R, Paull GC, Wilson RJ, Mill J, Santos EM (2016) Bisphenol A causes reproductive toxicity, decreases dnmt1 transcription, and reduces global DNA methylation in breeding zebrafish (Danio rerio). Epigenetics 11:526–538. doi: 10.1080/15592294.2016.1182272 CrossRefGoogle Scholar
- Nicodemus-Johnson J, Naughton KA, Sudi J, Hogarth K, Naurekas ET, Nicolae DL, Sperling AI, Solway J, White SR, Ober C (2016) Genome-wide methylation study identifies an IL-13-induced epigenetic signature in asthmatic airways. Am J Respir Crit Care Med 193:376–385. doi: 10.1164/rccm.201506-1243OC CrossRefGoogle Scholar
- Qiu W, Baccarelli A, Carey VJ, Boutaoui N, Bacherman H, Klanderman B, Rennard S, Agusti A, Anderson W, Lomas DA, DeMeo DL (2012) Variable DNA methylation is associated with chronic obstructive pulmonary disease and lung function. Am J Respir Crit Care Med 185:373–381. doi: 10.1164/rccm.201108-1382OC CrossRefGoogle Scholar
- Robbins SLC, Ramzi S (2004) Pathological basis of disease. Saunders, PhiladelphiaGoogle Scholar
- Stefanowicz D, Hackett TL, Garmaroudi FS, Günther OP, Neumann S, Sutanto EN, Ling KM, Kobor MS, Kicic A, Stick SM, Paré PD, Knight DA (2012) DNA methylation profiles of airway epithelial cells and PBMCs from healthy, atopic and asthmatic children. PLoS ONE 7:e44213. doi: 10.1371/journal.pone.0044213 CrossRefGoogle Scholar