Regulation of miR-486-3p on Cell Apoptosis of Breast Cancer Cell Line MCF-7
GAN Lin, WANG Yadong, WANG Ting, MIN Jie, LIAO Denghui, LÜ Gang*
To investigate the regulatory effect of miR-486-3p on apoptosis in breast cancer cells MCF-7 and its related mechanism, the expressions of miR-486-3p and apoptosis-related proteins in breast cancer tissues and adjacent normal tissues from 24 breast cancer patients were examined employing qRT-PCR and Western blot, respectively. miR-486-3p expression in breast cancer cells MCF-7, HBL101 and normal breast cells MCF10A was assayed by qRT-PCR. The groups of normal control, mimics control and miR-486-3p mimics, inhibitor control and miR-486-3p inhibitor were set in the experiments. After cell culture and transfection, cell proliferation was tested by CCK-8. Cell migration and invasion were assessed by wound healing and transwell assays. Cell apoptosis was evaluated by flow cytometry. The mRNA and protein expressions of Bcl-2, Bax and Caspase 3 were measured by qRT-PCR and Western blot. The results showed that in normal controls, compared with adjacent normal tissues, miRNA-486-3p expression in breast cancer tissues was significantly decreased (P<0.05). Bcl-2 protein level was remarkably increased (P<0.05), but Bax and caspase-3 protein levels were markedly reduced in breast cancer tissues (P<0.05). Compared with normal breast cells MCF10A, miR-486-3p expression was obviously reduced in breast cancer cells MCF-7 and HBL101 (P<0.05). miR-486-3p expression was lowest in MCF-1 cells. In the miRNA-486-3p mimic group, miR-486-3p expression was higher compared to the control group in MCF-7 and HBL101 cells (P<0.05). The optical density values of MCF-7 and HBL101 cells were significantly declined at 24 h, 48 h and 72 h (P<0.05). Moreover, MCF-7 cells had wider scratches (P<0.05). There were fewer transmembrane cells (P<0.05) and more apoptotic cells of MCF-7 cells (P<0.05). Bcl-2 level in MCF-7 cells was lower (P<0.05), but the levels of Bax and Caspase-3 were higher (P<0.05). In the miR-486-3p inhibitor group, miR-486-3p expression was decreased compared to the normal group in MCF-7 and HBL101 cells (P<0.05). The optical density values
of MCF-7 and HBL101 cells were significantly higher at 24 h, 48 h and 72 h (P<0.05). Additionally, MCF-7 cells had narrower scratches (P<0.05). There were more transmembrane cells (P<0.05) and fewer apoptotic cells of MCF-7 cells (P<0.05). Furthermore, Bcl-2 expression was obviously increased (P<0.05), but the levels of Bax and Caspase-3 were reduced in MCF-7 cells (P<0.05). Overall, these results indicated that miR-486-3p served as an antioncogene in breast cancer that might promote breast cancer apoptosis by regulating the mRNA and protein levels of Bcl-2, Bax and Caspase-3 related to apoptosis.
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