Fig. 5: NEAT1_1 upregulates AKR1C1 by sponging miR-338-3p in LUAD cells in vitro.

A Venn diagram showing the potential miRNAs which regulates the expression of AKRC1, predicted by 4 algorithms (PITA, miRmap, miRanDa, miRWalk, and DIANA microT). B The expressions of miR-338-3p and miR-185-5p in gefitinib-resistant and matched parental LUAD cells, normalized to U6. C Predicted sites on 3’-UTR of AKR1C1 mRNA to bind with miR-338-3p and matched established mutant sequences. D Luciferase reporter assay shows the luciferase activity of the LUAD cells transfected with AKR1C1 mRNA-3’-UTR-WT and AKR1C1 mRNA-3’-UTR-MUT after co-transfected with miR-338-3p mimic or miRNA mimic NC. E Overlapping lncRNAs of GEO datasets GSE199627 and GSE169513. F The expression of NEAT1 in gefitinib-resistant cell line and matched parental cells. G The subcellular location of NEAT1_1 (red) and miR-338-3p (green) in PC9/GR and PC9 cells, assayed by FISH. Scale bars, 10 μm. H The expressions of NEAT1_1 and NEAT1_2 in PC9 and PC9/GR cells, detected by qRT-PCR. I Predicted sites on NEAT1_1 to bind with miR-338-3p and matched established mutant sequences. J Luciferase reporter assay shows the luciferase activity of the PC9 cells transfected with NEAT1_1-WT and NEAT1_1-MUT after co-transfected with miR-338-3p mimic or miRNA mimic NC. K Enrichment level of NEAT1_1 and miR-338-3p in the Ago2 and IgG pellets of PC9 cells, respectively. L, M NEAT1_1/miR-338-3p/AKR1C1 axis was confirmed by performing miRNA rescue experiments in PC9/GR and PC9 cells. *P < 0.05; **P < 0.01; ***P < 0.001.