Decreased proliferation and erythroid differentiation of K562 cells by siRNA-induced depression of OCTN1 (SLC22A4) transporter gene
Decreased proliferation and erythroid differentiation of K562 cells by siRNA-induced depression of OCTN1 (SLC22A4) transporter gene
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DOI：
10.1007/s11095-007-9290-8
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发表时间：
2007-09-01
影响因子：
3.7
通讯作者：
Tamai, Ikumi
中科院分区：
文献类型：
--
作者：
Nakamura, Toshimichi;Sugiura, Shigeki;Tamai, Ikumi
最近，有报道OCTN1转运蛋白(SLC22A4)与类风湿性关节炎(RA)和克罗恩病有关。此外，我们报道了OCTN1在造血细胞中表达，优先在红系细胞中表达。因此，我们通过siRNA方法检测OCTN1在血细胞中的下调作用来评估OCTN1的生理作用。利用基于载体的短发夹状RNA(ShRNA)建立了OCTN1表达稳定降低的K562细胞系。通过细胞增殖、细胞分化和摄取作为OCTN1良好底物的麦角硫蛋白来研究OCTN1在K562细胞中的下调特征。与对照载体转染的细胞相比，它们的生长速度和向红细胞的丁酸依赖分化速度都有所下降。此外，K562细胞对[H-3]麦角硫蛋白的摄取表明，与OCTN1相似的钠离子依赖和高亲和力转运蛋白具有功能。此外，与野生型K562细胞相比，OCTN1表达降低的K562细胞对麦角硫蛋白的摄取减少，提示OCTN1参与了对细胞增殖和红系分化至关重要的生理物质的运输。
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.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.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 [H-3]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.It was suggested that OCTN1 is involved in the transport of physiological compounds that are important for cell proliferation and erythroid differentiation.