Expression, Localization and Protein-protein Interaction Analysis of lncRNA BDNF-AS Under Oxygen and Glucose Deprivation/Reoxygenation Condition
1. School of Basic Medical Sciences, Changsha Medical University, Changsha 410219, Hunan, China; 2. School of Computer Science and Engineering, Central South University, Changsha 410083, Hunan, China
Abstract：Abstract: Cerebral ischemia and hypoxia can cause the change of long non-coding RNA (lncRNA) expression. To investigate the expression and localization of lncRNA BDNF-AS and its interacting proteins under the con-dition of oxygen and glucose deprivation/reoxygenation (OGD/R), SH-SY5Y cells were treated with hypoxia for 8 h and reoxygenation (R) for 24 h to establish a OGD/R cell model. The cell viability was measured by Cell Counting Kit-8 (CCK-8) assay. The expression level of lncRNA-BDNF-AS was detected by qRT-PCR in both the nucleus and cytoplasm. The interacting proteins of lncRNA BDNF-AS were analyzed by using pull-down method and mass spectrometry. Functional analysis of interacting proteins was performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The protein-protein interac-tion network was analyzed using STRING database. Under OGD/R condition, the expression of lncRNA BDNF-AS was significantly increased, and was significantly higher in the cytoplasm than in the nucleus. In the OGD/R group, there were 120 kinds of specific proteins expressed in SH-SY5Y cells. Bioinformatic analysis showed that lncRNA BDNF-AS might interact with proteins including UBA52, NKAP, TBK1, RAB1A and RPL38. The-refore, lncRNA BDNF-AS may affect neuronal function by binding autophagy- and apoptosis-associated proteins.
