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. 2020 Jun 18;11(6):472. doi: 10.1038/s41419-020-2581-2

Fig. 4. DLEU2 functions as a competing endogenous RNA and regulates PIK3CD expression by sponging miR-30c-5p in LSCC cells.

Fig. 4

a The mRNA expression of miR-30c-5p was measured by qRT-PCR analysis in 66 LSCC specimens and matched adjacent normal tissues. b Spearman’s correlation analysis was used to evaluate the relationship between mRNA expression of DLEU2 and miR-30c-5p in 66 LSCC specimens. c After DLEU2 was overexpressed or knocked down in LSCC cells, qRT-PCR analysis was performed to measure the levels of miR-30c-5p mRNA. β-actin was used as an internal control. d, e The putative miR-30c-5p binding sites in the lncRNA DLEU2 (d) and the PIK3CD (e) sequences (PI3KCD-WT), as predicted by TargetScan, and the mutant sequences designed (lncRNA DLEU2-MUT and PIK3CD-MUT). Luciferase activity was determined by a luciferase reporter assay to confirm the direct correlation among the levels of miR-30c-5p, DLEU2, and PIK3CD. f QRT-PCR was performed to measure the PIK3CD expression of the indicated cells. g Spearman’s correlation analysis was used to evaluate the relationship between miR-30c-5p and PIK3CD mRNA expression in 66 LSCC specimens. h, i DLEU2 overexpression increased the protein (h) and mRNA (i) expression levels of PIK3CD. This effect was reversed by the induction of miR-30c-5p expression in LSCC cells. *P < 0.05, **P < 0.01, ***P < 0.001.