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| The inhibition of proliferation, migration and invasion of cervical cancer cells by miR-331-3p targeting NRP2 |
| 1.Jiulongpo District People's Hospital, Chongqing, 401329；2.The First Affiliated Hospital of Chongqing Medical University |
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Abstract To explore the regulative effect of miR-331-3p targeting NRP2 for the proliferation, migration, and invasion of cervical cancer HeLa cells.Methods: The cervical cancer HeLa cells were divided into control group, mimic NC group, miR-331-3p mimic group, siRNA NC group, si-NRP2 group, and miR-331-3p mimic+NRP2 group.The cells in mimic NC group and miR-331-3p mimic group were transfected with mimic NC plasmid and miR-331-3p mimic plasmid, respectively.The cells in siRNA NC group and si-NRP2 group were transfected with siRNA NC and si-NRP2, respectively.The cells in miR-331-3p mimic+NRP2 group were transfected with miR-331-3p mimic and NRP2 over expression plasmids, respectively.The cells in control group were not transfected.Targetscan software was used to predict the target gene, the dual luciferase reporter gene experiment was used to verify the relationship between miR-331-3p and targeting NRP2, and qRT-PCR was used to detect the expressions of miR-331-3p and NRP2.The cell proliferation, migration, and invasion situations of the cells in these groups after transfection were detected by CCK-8, cell scratch, and Transwell.Western blot was used to detect the protein expressions of NRP2, PCNA, MMP-2, and MMP9.Results: Compared with those of the normal human cervical epithelial cells, the expression level of miR-331-3p of the cervical cancer HeLa cells had downregulated significantly, but the expression of NRP2 had increased significantly(P<0.05).The dual luciferase reporter gene experiment confirmed that miR-3313p directly targeted to act on NRP2.Compared with those of the cells in the control group and the mimic NC group, the expression level of miR-331-3p of the cells in the over expression miR-331-3p group had increased significantly, but the expression levels of NRP2 mRNA and protein had decreased significantly(P<0.05).Over expression of miR-331-3p or inhibition of NRP2 expression all could inhibit cell growth rate, reduce the number of invaded cells, and increase the width of scratches.Western blot results showed that over expression of miR-331-3p or inhibition of NRP2 expression could inhibit the expressions of PCNA, MMP-2 and MMP-9 proteins(all P<0.05).And over expression of NRP2 could partially reverse the inhibitory effects of miR-331-3p for cell proliferation, invasion and migration.Conclusion: miR-331-3p targeting NRP2 can inhibit the proliferation, migration, and invasion of cervical cancer cells.
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