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Epigenetic Regulation of Organic Anion Transporting Polypeptide 1B3 in Cancer Cell Lines

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ABSTRACT

Purpose

The expression of a multispecific organic anion transporter, OATP1B3/SLCO1B3, is associated with clinical prognosis and survival of cancer cells. The aims of present study were to investigate the involvement of epigenetic regulation in mRNA expression of a cancer-type variant of OATP1B3 (Ct-OATP1B3) in cancer cell lines.

Methods

The membrane localization and transport functions of Ct-OATP1B3 were investigated in HEK293 cells transiently expressing Ct-OATP1B3. DNA methylation profiles around the transcriptional start site of Ct-OATP1B3 in cancer cell lines were determined. The effects of a DNA methyltransferase inhibitor and siRNA knockdown of methyl-DNA binding proteins (MBDs) on the expression of Ct-OATP1B3 mRNA were investigated.

Results

5′-RACE identified the TSS of Ct-OATP1B3 in PK-8 cells. Ct-OATP1B3 was localized on the plasma membrane, and showed the transport activities of E217βG, fluvastatin, rifampicin, and Gd-EOB-DTPA. The CpG dinucleotides were hypomethylated in Ct-OATP1B3-positive cell lines (DLD-1, TFK-1, PK-8, and PK-45P) but were hypermethylated in Ct-OATP1B3-negative cell lines (HepG2 and Caco-2). Treatment with a DNA methyltransferase inhibitor and siRNA knockdown of MBD2 significantly increased the expression of Ct-OATP1B3 mRNA in HepG2 and Caco-2.

Conclusions

Ct-OATP1B3 is capable of transporting its substrates into cancer cells. Its mRNA expression is regulated by DNA methylation-dependent gene silencing involving MBD2.

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Abbreviations

5′-RACE:

5′-rapid amplification cDNA ends

ChIP:

chromatin immunoprecipitation

E217βG:

estradiol 17β-D-glucuronide

Gd-EOB-DTPA:

gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid

ICP-MS:

inductively coupled plasma mass spectrometry

LC-MS/MS:

liquid chromatography-tandem mass spectrometry

MBD:

methyl-DNA binding protein

OATP:

organic anion transporting polypeptide

ORF:

open reading frame

PCR:

polymerase chain reaction

SLC:

Solute carrier

T-DMR:

tissue-dependent differentially methylated region

TSS:

transcriptional start site

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank Yuko Shiono and Yuta Shibue for their excellent technical assistance.

This study was supported by the Japan Society for the Promotion of Science [Grant-in-Aid for Scientific Research (S) 24229002, Scientific Research (B) 23390034, and Grant-in-Aid for Challenging Exploratory Research 21659037].

Author information

Authors and Affiliations

  1. Laboratory of Molecular Pharmacokinetics Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan

    Satoki Imai, Ryota Kikuchi, Yuri Tsuruya, Sotaro Naoi & Hiroyuki Kusuhara

  2. Pharmacokinetics Research Laboratories, Dainippon Sumitomo Pharma Co., Ltd., Osaka, Japan

    Satoki Imai

  3. Laboratory of Plant Nutrition and Fertilizers Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan

    Sho Nishida

  4. Sugiyama Laboratory, RIKEN Innovation Center, Research Cluster for Innovation, RIKEN, 1-6 Suehiro-cho Tsurumi-ku, Yokohama City, Kanagawa, 230-0045, Japan

    Yuichi Sugiyama

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  1. Satoki Imai
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  2. Ryota Kikuchi
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  4. Sotaro Naoi
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Correspondence to Yuichi Sugiyama.

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Imai, S., Kikuchi, R., Tsuruya, Y. et al. Epigenetic Regulation of Organic Anion Transporting Polypeptide 1B3 in Cancer Cell Lines. Pharm Res 30, 2880–2890 (2013). https://doi.org/10.1007/s11095-013-1117-1

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  • DOI: https://doi.org/10.1007/s11095-013-1117-1

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