GATA6-AS1 via Sponging miR-543 to Regulate PTEN/AKT Signaling Axis Suppresses Cell Proliferation and Migration in Gastric Cancer

Figure 5

PTEN is the target gene of miR-543 in GC cells. (a) Venn diagram represented the overlap of miR-543 targets based on three algorithms (PicTar, TargetScan, and TarBase). 11 predicted miR-543 targets are TWIST1, DYNC1LI2, PTEN, HERPUD1, FNDC3B, FOXP1, ANKRD13C, BTBD3, CDH11, ZNF281, and COPS2. (b) The starBase was utilized to predict the potential binding site between miR-543 and PTEN-3-UTR. The sequence of PTEN-3-UTR-WT containing miR-543-binding site was mutated to obtain PTEN-3-UTR-Mut plasmid. (c) The level of PTEN in the indicated GC cells after the upregulation or depletion of GATA6-AS1 was assessed via qRT-PCR. (d) Western blot analysis was utilized to measure the protein level of PTEN in the transfected cells after the overexpression and downregulation of GATA6-AS1. (e) RIP assays were conducted to analyze the association of GATA6-AS1, miR-543, and PTEN in GC cells. (f) RNA pull-down assays verified the binding between miR-543 and PTEN. (g) Luciferase activity of PTEN-3-UTR was detected. Student’s -test was used in (c)–(e) and one-way ANOVA followed by Dunnett’s test for (f) and (g). . n.s.: no significance.

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