Figure 6.
MCM3AP-AS1 regulates SIRT1 expression through miR-138-5p. (a) SIRT1 contained a binding sequence for miR-138-5p. (b) Dual-luciferase reporter gene assay confirmed that miR-138-5p mimics reduced the luciferase activity of the wild-type SIRT1 reporter, but had no effect on the luciferase activity of the mutant SIRT1 reporter. (c, d) RIP assay showed that miR-138-5p could directly bind with SIRT1 mRNA. (e) The RNA pull-down assay was performed to validate the interaction between miR-138-5p and SIRT1 mRNA. (f) qRT-PCR showed that SIRT1 mRNA was significantly down-regulated in OA cartilage tissues. (g) There was a negative correlation between miR-138-5p and SIRT1 mRNA expression levels in OA cartilage tissues. (h) There was a positive correlation between MCM3AP-AS1 and SIRT1 mRNA expression levels in OA cartilage tissues. (i) Overexpression of MCM3AP-AS1 could significantly increase SIRT1 expression. After co-transfection of MCM3AP-AS1/miR-138-5p, SIRT1 expression was significantly reduced. ns.: not statistically significant
*P < 0.05, **P < 0.01, and ***P < 0.001.