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Decreased Proliferation and Erythroid Differentiation of K562 Cells by siRNA-induced Depression of OCTN1 (SLC22A4) Transporter Gene

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Abstract

Purpose

Recently, it was reported that OCTN1 transporter (SLC22A4) is associated with rheumatoid arthritis (RA) and Crohn’s disease. Additionally, we reported that OCTN1 is expressed in hematopoietic cells, preferentially in erythroid cells. Accordingly, we assessed the physiological role of OCTN1 by examining the effect of knockdown of OCTN1 in blood cells using siRNA method.

Materials and Methods

Vector-based short hairpin RNA (shRNA) was used to establish K562 cell line which shows stably decreased expression of OCTN1. The characteristic of knockdown of OCTN1 in K562 cells was investigated by cell proliferation, cell differentiation, and uptake of ergothioneine that is a good substrate of OCTN1.

Results

Several clones of K562 cells exhibited significantly reduced expression of OCTN1 mRNA and protein. They also showed a decreased growth rate and butyrate-dependent differentiation to erythrocytes compared with control-vector transfected cells. In addition, uptake of [3H]ergothioneine by K562 cells suggested that Na+-dependent and high-affinity transporter which is similar to the characteristics of OCTN1 is functional. Moreover, uptake of ergothioneine by K562 cells which exhibit decreased-expression of OCTN1 was decreased in comparison with wild type K562 cells.

Conclusions

It was suggested that OCTN1 is involved in the transport of physiological compounds that are important for cell proliferation and erythroid differentiation.

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Abbreviations

ET:

ergothioneine

FBS:

fetal bovine serum

G3PDH:

glyceraldehyde-3-phosphate dehydrogenase

NMG:

N-methyl-d-glucamine

OCTN:

organic cation/carnitine transporter

PBS:

phosphate-buffered saline

PCR:

polymerase chain reaction

RA:

rheumatoid arthritis

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Acknowledgments

This investigation was supported in part by a Grant in Aid for Scientific Research from The Ministry of Education, Culture, Sports, Science, and Technology, Japan.

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Authors and Affiliations

  1. Department of Molecular Biopharmaceutics, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamasaki, Noda, Chiba, 278-8510, Japan

    Toshimichi Nakamura, Daisuke Kobayashi, Kenji Yoshida, Tomoji Maeda & Ikumi Tamai

  2. Research Institute for Frontier Medicine, Nara Medical University, Nara, Japan

    Shigeki Sugiura

  3. GenoMembrane Inc., Yokohama, Japan

    Hikaru Yabuuchi

  4. Department of Anatomy, Nihon University School of Medicine, Tokyo, Japan

    Shin Aizawa

Authors
  1. Toshimichi Nakamura
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  2. Shigeki Sugiura
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  3. Daisuke Kobayashi
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  4. Kenji Yoshida
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  5. Hikaru Yabuuchi
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  6. Shin Aizawa
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  7. Tomoji Maeda
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  8. Ikumi Tamai
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Correspondence to Ikumi Tamai.

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Nakamura, T., Sugiura, S., Kobayashi, D. et al. Decreased Proliferation and Erythroid Differentiation of K562 Cells by siRNA-induced Depression of OCTN1 (SLC22A4) Transporter Gene. Pharm Res 24, 1628–1635 (2007). https://doi.org/10.1007/s11095-007-9290-8

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  • DOI: https://doi.org/10.1007/s11095-007-9290-8

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