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Generation of a Cell Line Selectively Producing Functionally Active OATP1B1 Transporter

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Abstract

The solute carrier organic anion transporter family member, OATP1B1, is one of the most important transporter proteins, which mediate penetration of many endogenous substances and xenobiotics into hepatocytes. A model system providing expression of the functional protein is needed to assess interaction of OATP1B1 with various substances. Based on the HEK293 cells, we obtained the HEK293-OATP1B1 cell line, constitutively expressing the SLCO1B1 gene encoding the OATP1B1 transporter. Expression of the SLCO1B1 gene was confirmed by real-time PCR analysis and Western blotting. Functionality of the transporter was assessed by the transport of atorvastatin, which is a substrate of OATP1B1. Cells of the resulting cell line, which selectively express the functionally active recombinant OATP1B1 transporter, can be used to study functions of the protein and to test drugs for being substrates, inducers, and inhibitors of OATP1B1, and to assess the risks of drug interactions.

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Abbreviations

EGFP:

enhanced green fluorescent protein

HEK293:

human embryonic kidney cell line

OATP:

organic anion transporting polypeptides

OCT:

organic cation transporters

SLC:

solute carrier transporter proteins

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Acknowledgments

The authors are grateful for help provided by the Center for Collective Use “Industrial Biotechnology”, Russian Academy of Sciences, and to I. D. Solov’eva for help in using confocal microscopy.

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Authors and Affiliations

  1. Bach Institute of Biochemistry, Federal Research Centre “Fundamentals of Biotechnology”, Russian Academy of Sciences, 119071, Moscow, Russia

    Mariia S. Kotliarova, Nonna I. Nadolinskaia, Mikhail V. Zamakhaev & Anna V. Goncharenko

  2. Pavlov Ryazan State Medical University, 390026, Ryazan, Russia

    Aleksei V. Shchulkin, Pelageya D. Erokhina, Pavel Y. Mylnikov & Elena N. Yakusheva

Authors
  1. Mariia S. Kotliarova
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  2. Aleksei V. Shchulkin
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  3. Pelageya D. Erokhina
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  4. Pavel Y. Mylnikov
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  5. Elena N. Yakusheva
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  6. Nonna I. Nadolinskaia
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  7. Mikhail V. Zamakhaev
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  8. Anna V. Goncharenko
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Contributions

A.V.G., A.V.Shch., concept and supervision of the study; M.S.K., P.D.E., P.Yu.M., A.V.Shch., E.N.Ya., M.V.Z., N.I.N., conducting experiments; P.D.E., P.Yu.M., A.V.Shch., E.N.Yy., M.S.K., A.V.G., writing text of the paper; M.V.Z., editing text of the paper.

Corresponding author

Correspondence to Anna V. Goncharenko.

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The authors declare no conflict of interests in financial or any other sphere. This article does not describe any studies involving humans or animals performed by any of the authors.

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Kotliarova, M.S., Shchulkin, A.V., Erokhina, P.D. et al. Generation of a Cell Line Selectively Producing Functionally Active OATP1B1 Transporter. Biochemistry Moscow 88, 1267–1273 (2023). https://doi.org/10.1134/S0006297923090067

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  • DOI: https://doi.org/10.1134/S0006297923090067

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