Fig. 7

ALDOA is a target of miR-329-3p. (A) The potential binding site for miR-329-3p in ALDOA-3′UTR was predicted. (B) Total RNAs were isolated from fresh clinical specimens. ALDOA levels in 20 paired clinical specimens were determined by real-time PCR (n = 20). GAPDH served as an internal control. (C) The cells were co-transfected with miR-329-3p mimic or NC mimic and wild-type or mutant ALDOA-3′UTR. The binding of miR-329-3p to ALDOA-3′UTR was verified by a dual-luciferase reporter assay (n = 3). (D) The cells were transfected with miR-329-3p mimic or inhibitor, and then ALDOA levels were assessed by real-time PCR (n = 3). GAPDH served as an internal control. (E) AGS cells stably knocked down PSMA3-AS1 were transfected with miR-329-3p inhibitor or NC inhibitor. HGC-27 cells stably overexpressing PSMA3-AS1 were transfected with miR-329-3p mimic or NC mimic. ALDOA levels in AGS cells were assessed by real-time PCR. GAPDH served as an internal control. (F) After transfection, cell proliferation capacity was measured by CCK-8 assay (n = 3). (G) After transfection, cell migration ability was evaluated by wound healing assay (n = 5). (H) After transfection, cell invasion ability was assessed by transwell invasion assay (n = 5). (I) After transfection, intracellular ROS level was examined using DCFH-DA fluorescent probes (n = 3). (J) After transfection, MDA content, SOD activity, and GSH-Px activity were measured (n = 3). MDA content is expressed as nmol/mg prot. SOD activity is expressed as U/mg prot. GSH-Px activity is expressed as U/mg prot. ^**P < 0.01 compared with Normal, NC mimic, NC inhibitor, sh-NC, or vector. ^#P < 0.05 and ^##P < 0.01 compared with sh-PSMA3-AS1 + NC inhibitor or miR-329-3p mimic + vector. Student′s t-test was used to compare two groups, and one-way ANOVA with Tukey′s post-hoc test was used to compare multiple groups