Pharmaceutical Research

, Volume 24, Issue 9, pp 1628–1635 | Cite as

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

  • Toshimichi Nakamura
  • Shigeki Sugiura
  • Daisuke Kobayashi
  • Kenji Yoshida
  • Hikaru Yabuuchi
  • Shin Aizawa
  • Tomoji Maeda
  • Ikumi Tamai
Research Paper

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.

Key words

ergothioneine erythrocyte K562 OCTN1 siRNA transporter 

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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Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Toshimichi Nakamura
    • 1
  • Shigeki Sugiura
    • 2
  • Daisuke Kobayashi
    • 1
  • Kenji Yoshida
    • 1
  • Hikaru Yabuuchi
    • 3
  • Shin Aizawa
    • 4
  • Tomoji Maeda
    • 1
  • Ikumi Tamai
    • 1
  1. 1.Department of Molecular Biopharmaceutics, Faculty of Pharmaceutical SciencesTokyo University of ScienceNodaJapan
  2. 2.Research Institute for Frontier MedicineNara Medical UniversityNaraJapan
  3. 3.GenoMembrane Inc.YokohamaJapan
  4. 4.Department of AnatomyNihon University School of MedicineTokyoJapan

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