The Department of Occupational and Environment Health, School of Public Health, Ningxia Medical University, 1160 Shengli Street, Xingqing District, Yinchuan 750004, People's Republic of China.
Shanghai Municipal Center for Disease Control & Prevention, Shanghai 200336, People's Republic of China.
Environ Toxicol Pharmacol. 2016 Mar;42:205-11. doi: 10.1016/j.etap.2016.01.018. Epub 2016 Jan 25.
Paraquat (PQ) exposure influences central nervous system and results in serious neurotoxicity in vitro and in vivo. However, the role of PQ exposure in the development of CNS remains unclear. In present study, we investigated microRNAs (miRNAs) expression profiling and cell differential status following PQ treatment in human neural progenitor cells (hNPCs) as well as involved mechanism. Microarray profiling of miRNAs expression of PQ treated cell line and their corresponding control was determined. Differentially expression miRNAs were confirmed by quantitative real time PCR. Neural cell differentiation was performed with immunocytochemical analysis. Predicated target of miRNA was identified with luciferase reports and quantitatively analyzed using western blotting. Our results found PQ dramatically suppressed neural cell differentiation ability. 43 differentially expressed miRNAs were identified in PQ treated cells. The expression levels were over expressed in 25 miRNAs, whereas 18 miRNAs were suppressed. More importantly, we observed that miR-200a expression level to be lower in PQ treated cells. Luciferase assay and protein expression results confirmed the direct binding effect between CTNNB1 and miR-200a following PQ exposure. Collectively, our data suggested that down regulation of miR-200a in the PQ treated neural stem cell significantly participated in the differentiation processes and subsequently resulting in decreased cell viability, increased epithelial-mesenchymal transition process and the inhibited differential through CTNNB1 pathway.
百草枯（PQ）暴露会影响中枢神经系统，并在体外和体内导致严重的神经毒性。然而，PQ 暴露在中枢神经系统发育中的作用尚不清楚。在本研究中，我们研究了 PQ 处理后人类神经祖细胞（hNPC）中 microRNAs（miRNAs）表达谱和细胞差异状态以及相关机制。通过 miRNA 表达的微阵列分析确定 PQ 处理细胞系及其相应对照。通过定量实时 PCR 确认差异表达的 miRNAs。通过免疫细胞化学分析进行神经细胞分化。使用荧光素酶报告鉴定 miRNA 的预测靶标，并使用 Western blot 进行定量分析。我们的结果发现 PQ 显著抑制了神经细胞的分化能力。在 PQ 处理的细胞中鉴定出 43 个差异表达的 miRNAs。在 25 个 miRNA 中表达水平上调，而 18 个 miRNA 被抑制。更重要的是，我们观察到 PQ 处理细胞中的 miR-200a 表达水平较低。荧光素酶测定和蛋白表达结果证实了 PQ 暴露后 CTNNB1 和 miR-200a 之间的直接结合效应。总之，我们的数据表明 PQ 处理的神经干细胞中 miR-200a 的下调显著参与了分化过程，随后导致细胞活力降低、上皮-间充质转化过程增加以及通过 CTNNB1 途径抑制分化。
