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. 2021 Mar 17;11:605884. doi: 10.3389/fonc.2021.605884

Figure 6.

Figure 6

LBX1-AS1/miR-182-5p axis is critical for FOXO3 expression. (A) Bioinformatics analysis revealed the predicted binding sites between FOXO3 and miR-182-5p. (B) Luciferase reporter assay demonstrated miR-182-5p mimics significantly decreased the luciferase activity of FOXO3-WT in SCC-4 cells. (C) The transfection efficiency of miR-182-5p mimics and inhibitor in SCC-4 and CAL-27 cells. (D, E) The mRNA (D) and protein (E) level of FOXO3 was detected through qRT-PCR and western blotting after transfection with miR-182-5p mimics and inhibitor in SCC-4 and CAL-27 cells. (F) LBX1-AS1 overexpression upregulated FOXO3 and siRNA downregulated FOXO3, this effect can be reversed by co-transfection with miR-182-5p mimics or miR-182-5p inhibitors (miR-182-5p-inh) in SCC-4 or CAL-27 cells respectively. (G) The expression levels of FOXO3 in 40 OSCC tissues and matched para-carcinoma normal tissues was determined by qRT-PCR. (H) Expression levels of FOXO3 negatively correlated with miR-182-5p in OSCC tissues. (I) Expression levels of FOXO3 positively correlated with LBX1-AS1 in OSCC tissues. **, ##P < 0.01 and ***, ###P < 0.001 as indicated. * vs. miR-NC; Control, # vs. in-NC.