ABSTRACT
Aim To investigate the regulatory role and mechanism of resveratrol in inhibiting autophagy and promoting apoptosis in choroidal melanoma cells. Methods Choroidal melanoma cells (MUM2B) were divided into control and experimental groups, and treated with different concentrations of resveratrol (0, 10, 20,40,60,80 μmol ·L
ABSTRACT
Objective:This study aims to investigate the expression pattern of miR-512-3p in breast cancer and noncancerous paired specimens as well as the effects of miR-512-3p on the proliferation, apoptosis, cell cycle, and cloning of MD-MBA-231 breast cancer cells. The study also aims to identify the miR-512-3p target gene. Methods:Reverse transcription-polymerase chain reaction (RT-PCR) was conducted to quantify the miR-512-3p expression in breast cancer and noncancerous paired specimens. Methylthiazol tetrazolium (MTT) assay, flow cytometry, and clone formation assay were used to characterize the function of miR-512-3p in breast cancer. Target prediction was performed using the TargetScan, PicTar, and miRanda software. The results were validated using RT-PCR and western blot target validation. Results:The relative expression of miR-512-3p in breast cancer specimens was significantly lower than those in normal breast specimens (P<0.05). MTT assay revealed that 48 h after transfection, miR-512-3p significantly repressed the proliferation of MD-MBA-231 cells at a suppression rate of 45.38%and at a concentration of 100 nmol/L. MiR-512-3p increased the percentage of early apoptotic cells in the treatment groups (9.32 ± 0.41)%compared with those in the blank controls (3.1 ± 0.54)%and in the negative controls (2.9 ± 0.39)%(P<0.05). Significant differences were found in the percentages of the G0/G1-and G2/M-phase cells after miR-512-3p transfection compared with those in the controls (P<0.05). In the cloning assay, clone formation was inhibited in the miR-512-3p-transfected groups compared with those in the control groups. RT-PCR and western blot results indicate that miR-512-3p significantly inhibited the c-FLIP mRNA and protein expression. Conclusion:MiR-512-3p expression is relatively decreased in breast cancer specimens compared with those in the normal samples. The negative effect of miR-512-3p on cell proliferation and clone formation and its positive effect on early apoptosis through c-FLIP targeting suggest that miR-512-3p acts as a tumor suppressor gene in breast cancer. Therefore, miR-512-3p may be a new target for the diagnosis and treatment of breast cancer in the future.