Knockdown of long noncoding RNA WNT5A-AS restores the fate of neural stem cells exposed to sevoflurane via inhibiting WNT5A/Ryk-ROS signaling

Abstract

Long noncoding RNAs (lncRNAs) have been implicated in neurogenesis. LncRNA WNT5A-AS is upregulated in neural stem cells (NSCs), the proliferation of which is inhibited by sevoflurane. Thus, we hypothesized that knocking down of lncRNA WNT5A-AS may restore the fate of NSCs exposed to sevoflurane. To test this hypothesis, NSCs obtained from postnatal Sprague-Dawley rats were exposed to 2.4% sevoflurane or control gas for 6 h. Bioinformatics analysis, quantitative PCR and RNA interference technology were used to identify the properties of lncRNA WNT5A-AS. Cell proliferation was assessed using counting a Cell Counting Kit-cell 8 assay, a 5-ethynyl-2'-deoxyuridine incorporation assay, and a plate cloning assay. Cell survival was detected by flow cytometry, which was also used to examine the levels of reactive oxygen species (ROS) and the cell cycle. The levels of WNT5A and receptor tyrosine kinase (Ryk) were measured via Western blotting. LncRNA WNT5A-AS was identified to have low coding potency and to be located on the antisense strand of WNT5A. The level of upregulated lncRNA WNT5A-AS was positively correlated with that of WNT5A in response to sevoflurane exposure. The knockdown of lncRNA WNT5A-AS promoted the proliferation and survival of NSCs, whereas it suppressed the WNT5A/Ryk-ROS signaling and drove cell cycle processes. Taken together, findings strongly suggest that the inhibition of lncRNA WNT5A-AS can rescue the fate of NSCs. In addition, WNT5A/Ryk-ROS signaling might be a downstream target of lncRNA WNT5A-AS.

Keywords: Long noncoding RNA; Neural stem cells; Proliferation; Sevoflurane; WNT5A.