Figure 1.

miR-433 is associated with cell growth inhibition induced by high glucose, and miR-433 negatively regulates COX2 expression in Min-6 pancreatic β cells. (A) CCK-8 assay was used to determine cell viability in Min-6 cells treated with either a low (5 mmol/l) or a high (11 mmol/l) concentration of glucose for 24 h. (B) Expression levels of miR-433, miR-199-5p, miR-30a and miR-22 were analyzed by RT-qPCR in Min-6 cells treated with either 5 mmol/l or 11 mmol/l glucose. (C) Bioinformatics prediction of miR-433 binding site in the 3′-UTR of COX2 mRNA. (D) HEK293T cells were co-transfected with a WT or a MUT COX 3′-UTR reporter plasmid along with miR-433 mimics or NC mimics. Activity was measured by dual luciferase reporter assay and presented as relative luciferase activity. (E) COX2 mRNA expression levels were detected by RT-qPCR in Min-6 cells cultured in high glucose conditions following transfection with miR-433 mimics. (F) mRNA levels of COX2 were determined by RT-qPCR in Min-6 cells treated with 5 mmol/l or 11 mmol/l glucose. Data are expressed as the mean ± standard deviation; **P<0.01 and ***P<0.001 vs. low-glucose treatment or NC. CCK-8, Cell Counting Kit-8; COX2, cyclooxygenase 2; miR, microRNA; miRNA, microRNA; Mmu, Mus musculus; MUT, mutant; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; UTR, untranslated region; WT, wild-type.