Fig 2: Circ_0008450 regulated AKAP1 expression by miR-431 in hypoxia-disposed SNU-387 and Huh7 cells. (A and B) Effect of AKAP1 enhancement or (C and D) miR-431 inhibitor on circ_0008450 depletion-mediated expression of AKAP1 mRNA and protein of hypoxia-disposed SNU-387 and Huh7 cells was examined using reverse transcription-quantitative PCR and western blotting. *P<0.05. Circ, circular; AKAP1, A-kinase anchor protein 1; miR, microRNA; si-, small interfering; NC, negative control.

Fig 3: miR-431 modulated viability, apoptosis and glycolysis of hepatocellular cancer cells via AKAP1 under hypoxia treatment. The introduction of pc-AKAP1 on the influence of miR-431 mimic-mediated expression of AKAP1 (A) mRNA and (B) protein of hypoxia-exposed SNU-387 and Huh7 cells was determined with reverse transcription-quantitative PCR and western blotting, respectively. The enhancement of AKAP1 on the impacts of miR-431 mimic-mediated (C) viability, (D) apoptosis and (E and F) glycolysis of hypoxia-disposed SNU-387 and Huh7 cells were evaluated using Cell Counting kit-8, flow cytometry and glycolysis assays. *P<0.05. miR, microRNA; AKAP1, A-kinase anchor protein 1; H, hypoxic; NC, negative control.

Fig 4: AKAP1 was a target of miR-431 in hepatocellular cancer cells. (A) The binding sites between miR-431 and AKAP1 were predicted using the Starbase database. Dual-luciferase reporter assays were conducted for the evaluation of the luciferase activity in 293T cells contransfected with WT or MUT reporter vectors of AKAP1 and (B) miRNA NC, miR-431 mimic, (C) inhibitor NC or miR-431 inhibitor. Effect of miR-431 on expression of AKAP1 (D) mRNA and (E) protein of SNU-387 and Huh7 cells was analyzed by RT-qPCR and western blot analysis, respectively. Under hypoxic conditions, the (F) mRNA and (G) protein levels of AKAP1 in SNU-387 and Huh7 cells were detected by RT-qPCR and western blot analysis, respectively. *P<0.05. AKAP1, A-kinase anchor protein 1; miR, microRNA; WT, wild-type; MUT, mutant; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; UTR, untranslated region.

Fig 5: Effect of circ_0008450 on tumor growth in vivo. (A) On day 7, the tumor volumes of the mice in the sh-circ_0008450 and sh-NC groups were measured every 4 days. (B) Tumor weights were assessed on day 27. Expression of (C) circ_0008450 and (D) miR-431 in tumors were detected via RT-qPCR. Expression of AKAP1 (E) mRNA and (F) protein in tumors were examined by RT-qPCR and western blotting, respectively. *P<0.05. Circ, circular; sh, short hairpin; NC, negative control; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR; AKAP1, A-kinase anchor protein 1.