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SMS regulates the expression and function of P-gp and MRP2 in Caco-2 cells

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Abstract

Sphingomyelin synthase (SMS) has two isoforms of SMS1 and SMS2, the last enzyme involved in the biosynthesis of sphingomyelin (SM), and has impact on the expression of membrane proteins. In the present study, we explored the potential effects of SMS on drug transporters, a special family of membrane proteins in human intestinal epithelial Caco-2 cells. The specific knockdown of SMS1 or SMS2 with siRNA in Caco-2 cells substantially decreased the expression and function of P-glycoprotein (P-gp) and multidrug resistance protein 2 (MRP2) rather than other drug transporters MRP1, MRP3, PEPT1, OATP2B1, and BCRP. In the SMS1 stable overexpressed Caco-2 cell line, the expression levels of P-gp and MRP2 and transcription factor pregnane X receptor (PXR) were upregulated and the phosphorylation levels of signaling pathways janus protein tyrosine kinase 2 (JAK-2) and extracellular signal-regulated kinases (ERK) were also evidently increased; however, the upregulated mRNA expression levels of PXR, P-gp, and MRP2 were diminished by inhibiting the phosphorylation of ERK and JAK-2. Furthermore, the SMS1 overexpression in Caco-2 cells altered the expression levels of ERM proteins ezrin and moesin, which are closely connected to the function of drug transporters. In conclusion, we herein demonstrate for the first time that in Caco-2 cells SMS regulates the expression and function of drug transporters P-gp and MRP2, and their regulator PXR is mediated by phosphorylated ERK and JAK-2 signaling pathways.

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Abbreviations

ABC:

ATP-binding cassette

ERK:

Extracellular signal-regulated kinases

ERM:

Ezrin, radixin, and moesin

JAK-2:

Janus protein tyrosine kinase 2

MDR:

Multidrug resistance

MRP2:

Multidrug resistance protein 2

P-gp:

P-glycoprotein

PXR:

Pregnane X receptor

SLC:

Solute carrier

SM:

Sphingomyelin

SMS:

Sphingomyelin synthase

TLC:

Thin layer chromatography

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81373396), the Key Biomedical Program of Shanghai (No. 12431900204), the Research Fund for the Doctoral Program of Higher Education of China (No. 20130071110037), and the Research Fund of the State Key Laboratory of Genetic Engineering, Fudan University.

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    Authors and Affiliations

    1. State Key Laboratory of Genetic Engineering, Department of Biochemistry, School of Life Sciences, Fudan University, Songhu Road 2005, Shanghai, 200438, China

      Guiying Jin, Yang Li, Yuwen Zhu, Lisha Du & Qing Yang

    2. Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

      Junkai Yan & Qing Yang

    3. Shanghai Engineering Research Center of Industrial Microorganisms, Songhu Road 2005, Shanghai, 200438, China

      Qing Yang

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    1. Guiying Jin
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    Correspondence to Qing Yang.

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    Guiying Jin and Yang Li contributed equally to this work.

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    Jin, G., Li, Y., Zhu, Y. et al. SMS regulates the expression and function of P-gp and MRP2 in Caco-2 cells. Cell Biol Toxicol 32, 483–497 (2016). https://doi.org/10.1007/s10565-016-9348-7

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