Bezerra D P, Keasey M, Oliveira J R M
Keizo Asami Laboratory, Federal University of Pernambuco, Recife, PE, Brazil.
Department of Biomedical Sciences - Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
J Mol Neurosci. 2017 May;62(1):28-33. doi: 10.1007/s12031-017-0906-0. Epub 2017 Mar 16.
PiT1 (SLC20A1) and PiT2 (SLC20A2) are members of the mammalian type-III inorganic phosphate transporters and recent studies linked SLC20A2 mutations with primary brain calcifications. MicroRNAs (miRNAs) are endogenous noncoding regulatory RNAs and MicroRNA-9 (miR-9) modulates neurogenesis but is also involved with different types of cancer. We evaluated possible interactions between miR-9 and the phosphate transporters (PiT1 and PiT2). SLC20A2, platelet-derived growth factor receptor beta (PDGFRB) and Fibrillin-2 (FBN2) showed binding sites with high affinity for mir-9, In silico. miR-9 mimic was transfected into HEK293 cells and expression was confirmed by RT-qPCR. Overexpression of miR-9 in these cells caused a significant reduction in PiT2 and FBN2. PDGFRB appeared to be decreased, but was not significantly down-regulated. PiT1 showed no significant difference relative to controls. The down-regulation of PiT2 protein by miR-9 was confirmed by western blotting. In conclusion, we showed that miR-9 can down-regulate PiT2, in HEK293 cells. [corrected].
PiT1（溶质载体家族20成员1，SLC20A1）和PiT2（溶质载体家族20成员2，SLC20A2）是哺乳动物III型无机磷酸盐转运蛋白家族的成员，最近的研究将SLC20A2突变与原发性脑钙化联系起来。微小RNA（miRNA）是内源性非编码调节RNA，微小RNA-9（miR-9）可调节神经发生，但也与不同类型的癌症有关。我们评估了miR-9与磷酸盐转运蛋白（PiT1和PiT2）之间可能的相互作用。在计算机模拟中，SLC20A2、血小板衍生生长因子受体β（PDGFRB）和原纤蛋白-2（FBN2）显示出与miR-9具有高亲和力的结合位点。将miR-9模拟物转染到HEK293细胞中，并通过逆转录定量聚合酶链反应（RT-qPCR）确认其表达。在这些细胞中miR-9的过表达导致PiT2和FBN2显著减少。PDGFRB似乎有所下降，但未显著下调。与对照组相比，PiT1没有显著差异。通过蛋白质印迹法证实了miR-9对PiT2蛋白的下调作用。总之，我们表明在HEK293细胞中，miR-9可以下调PiT2。[已校正]
