Protein 4.1R is Involved in the Transport of 5-Aminolevulinic Acid by Interaction with GATs in MEF Cells
Protein 4.1R is Involved in the Transport of 5-Aminolevulinic Acid by Interaction with GATs in MEF Cells
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蛋白 4.1R 通过与 MEF 细胞中的 GAT 相互作用参与 5-氨基乙酰丙酸的转运
DOI：
10.1111/php.12842
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发表时间：
2018
影响因子：
3.3
通讯作者：
Ji Zhenyu
中科院分区：
文献类型：
--
作者：
Ning Shuwei;Kang Qiaozhen;Fan D;an;Liu Jingjing;Xue Chaoyue;Zhang Xiaolin;Ding Cong;Zhang Jianying;Peng Qian;Ji Zhenyu
基于5-氨基乙酰丙酸(5-ALA)的光动力疗法(PDT)已成功地用于癌症的治疗。然而，5-丙氨酸转运到癌细胞的机制仍不完全清楚。已有研究证实，膜骨架蛋白4.1R可能影响5-ALA-PDT的效率。在这项研究中，我们研究了4.1R在5-ALA进入细胞的运输中的作用。将野生型(4.1R+/+)和4.1R基因敲除(4.1R−/−)的小鼠胚胎成纤维细胞分别与1mm5-ALA和不同浓度的转运蛋白特异性抑制剂GAT(1-3)共同孵育。我们的结果表明，GAT1和GAT2的抑制显著地减弱了细胞内PpIX的产生，活性氧(ROS)水平和5-ALA诱导的光损伤。而对GAT3的抑制则没有表现出这种作用。进一步研究表明，4.1R+/+−/−MEF细胞中GAT1和GAT2的表达低于4.1R+/+MEF细胞。此外，4.1R直接与GAT1和GAT2结合。总之，GAT1和GAT2转运体参与了MEF细胞对5-ALA的摄取。4.1R通过至少部分地与5-ALA-PDT中的GAT1和GAT2转运蛋白相互作用，在5-ALA-PDT转运5-ALA中发挥重要作用。
5‐aminolevulinic acid (5‐ALA)‐based photodynamic therapy (PDT) has been successfully used in the treatment of cancers. However, the mechanism of 5‐ALA transportation into cancer cells is still not fully elucidated. Previous studies have confirmed that the efficiency of 5‐ALA‐PDT could be affected by the membrane skeleton protein 4.1R. In this study, we investigated the role of 4.1R in the transport of 5‐ALA into cells. Wild‐type (4.1R+/+) and 4.1R gene knockout (4.1R−/−) mouse embryonic fibroblast (MEF) cells were incubated with 1 mm5‐ALA and different concentrations of specific inhibitors of GABA transporters GAT (1‐3). Our results showed that the inhibition of GAT1 and GAT2 in particular markedly attenuated the intracellular PpIX production, reactive oxygen species (ROS) level and 5‐ALA‐induced photodamage. However, the inhibition of GAT3 did not show such effects. Further research showed that 4.1R−/−MEF cells had a lower expression of GAT1 and GAT2 than 4.1R+/+MEF cells. Additionally, 4.1R directly bound to GAT1 and GAT2. Taken together, GAT1 and GAT2 transporters are involved in the uptake of 5‐ALA in MEF cells. 4.1R plays an important role in transporting 5‐ALA into cells via at least partly interaction with GAT1 and GAT2 transporters in 5‐ALA‐PDT.