miR-29a Negatively Affects Glucose-Stimulated Insulin Secretion and MIN6 Cell Proliferation via Cdc42/β-Catenin Signaling
Effects of miR-29a/Cdc42 on MIN6 cells. (a) Effects of miR-29a on Cdc42 protein expression. Western blot to detect Cdc42 protein expression after transfection with miR-29a mimic and inhibitor (n = 3). , compared with miR-29a NC group, as assessed by paired Student’s t-test. (b) Bioinformatics analysis of the miR-29a binding site in the Cdc42 mRNA 3′-UTR. (c) Luciferase reporter assays indicate that miR-29a binds to Cdc42 mRNA in MIN6 cells. , compared with nonmutated Cdc42 3′UTR + miR-29a-NC group, as assessed by paired Student’s t-test. (d) Effects of miR-29a/Cdc42 on MIN6 cell proliferation. CCK assay to detect MIN6 cell proliferation after simultaneous overexpression of miR-29a and Cdc42, and after simultaneous interference with miR-29a and Cdc42 expression (n = 3). , compared with miR-29a mimic group, and , compared with miR-29a inhibitor group, as assessed by one-way ANOVA, followed by Fisher’s least significant difference test. (e) Effects of miR-29a/Cdc42 on insulin secretion by MIN6 cells. ELISA to detect insulin secretion by MIN6 cells after transient transfection with miR-29a mimic + Cdc42-pcDNA3.1 and miR-29a inhibitor + Cdc42-siRNA under high-glucose (20.0 mM) stimulation (n = 3). , compared with miR-29a inhibitor group, and , compared with miR-29a mimic group, as assessed by paired Student’s t-test. Data are shown as mean ± SD. NC: negative control; mmu: mouse lemur; 3p: mmu-miR-29a is produced from the 3′ end arm of the double strand of miR-29a; mut: mutated.
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