Abstract:

Objective To investigate the effect of microRNA-147（miR-147）on the expression of murine double minute 2（MDM2）and its effect on the proliferation and apoptosis of ovarian cancer cell line Skov3. Methods The Skov3 cells were transfected with miR-147 mimics（miR-147 mimics group）or negative control（NC group）by Lipo2000 liposome method. The real-time quantitative PCR（QPCR）method was used to detect the miR-147 level of Skov3 cells at 48 h post-transfection to evaluate the transfect efficiency. MTT method and flow cytometry were used to detect the proliferation and apoptosis of the two groups after transfection, respectively. The levels of MDM2， p53 and caspase-3 were measured by Western blotting. The dual luciferase reporter assay was employed to verify the relationship between miR-147 and target MDM2 and the binding site. Results QPCR results showed that the level of miR-147 in miR-147 mimics group was significantly higher than that in NC group，and the difference was statistically significant（P<0.05）， indicating that miR-147 was successfully over-expressed in Skov3 cells. Compared with NC group， the proliferative rate and MDM2 level of miR-147 mimics group were all decreased， but the apoptotic rate and levels of p53 and caspase-3 were all increased（P<0.05）. Dual luciferase reporter assay showed that miR-147 could significantly inhibit the luciferase activity of cells transfected with wild-type MDM2-3’（untranslated region， UTR） plasmid，and had no effect on the luciferase activity of cells transfected with mutant MDM2-3’UTR plasmid. Conclusion MiR-147 can target the expression of MDM2，and over-expression of miR-147 can inhibit the proliferation and induce apoptosis of Skov3 cells，and it has a certain reference value in the prevention and treatment of ovarian cancer.