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Deoxycholic acid inhibited proliferation and induced apoptosis and necrosis by regulating the activity of transcription factors in rat pancreatic acinar cell line AR42J

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Abstract

The objective of this study is to investigate the effect of deoxycholic acid (DCA) on rat pancreatic acinar cell line AR42J and the functional mechanisms of DCA on AR42J cells. AR42J cells were treated with various concentrations of DCA for 24 h and also treated with 0.4 mmol/L DCA for multiple times, and then, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to detect the AR42J cell survival rate. Flow cytometric was used to detect the cell apoptosis and necrosis in AR42J cells treated with 0.4 mmol/L and 0.8 mmol/L DCA. The cells treated with phosphate buffer saline (PBS) were served as control. In addition, the DNA-binding activity assays of transcription factors (TFs) in nuclear proteins of cells treated with DCA were determined using Panomics Procarta Transcription Factor Assay Kit. The relative survival rates were markedly decreased (P < 0.05) in a dose- and time-dependent manner. Compared with control group, the cell apoptosis and necrosis ratio were both significantly elevated in 0.4 mmol/L DCA and 0.8 mmol/L DCA groups (P < 0.01). A significant increase (P < 0.05) in the activity of transcription factor 2 (ATF2), interferon-stimulated response element (ISRE), NKX-2.5, androgen receptor (AR), p53, and hypoxia-inducible factor-1 (HIF-1) was observed, and the activity of peroxisome proliferator-activated receptor (PPAR), activator protein 1 (AP1), and E2F1 was reduced (P < 0.05). In conclusion, DCA inhibited proliferation and induced apoptosis and necrosis in AR42J cells. The expression changes of related genes regulated by TFs might be the molecular mechanism of AR42J cell injury.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (81303110) and the Specialized Research Fund for the Doctoral Program of Higher Education of China (20112105120007).

Authors’ contributions

GZ participated in the design of this study, and they both performed the statistical analysis. HC carried out the study, together with JZ, collected important background information, and drafted the manuscript. DS and BQ conceived of this study and participated in the design and helped to draft the manuscript. All authors read and approved the final manuscript.

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All authors declare that they have no conflict of interest to state.

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Correspondence to Hailong Chen.

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Editor: T. Okamoto

Guixin Zhang and Jingwen Zhang are co-first authors.

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Zhang, G., Zhang, J., Shang, D. et al. Deoxycholic acid inhibited proliferation and induced apoptosis and necrosis by regulating the activity of transcription factors in rat pancreatic acinar cell line AR42J. In Vitro Cell.Dev.Biol.-Animal 51, 851–856 (2015). https://doi.org/10.1007/s11626-015-9907-x

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  • DOI: https://doi.org/10.1007/s11626-015-9907-x

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