Targeted regulation of microRNA?186 on ARAP2 expression and its effect on invasion and migration of oral squamous cell carcinoma cells

LIU Rongguang; ZHANG Ying; YOU Huilin

doi:10.3969/j.issn.1009-0460.2019.08.005

Abstract：Objective To investigate the effect of microRNA-186 (miR-186) on the expression of A-kinase anchoring protein 2 (ARAP2) as well as the invasion and migration of oral squamous cell carcinoma ( OSCC) cells. Methods Real-time quantitative polymerase chain reaction (QPCR) was used to detect the miR-186 level in OSCC HN13 cells and human oral epithelial cells HOK. HN13 cells were transfected with miR-186 mimics ( Mimics group), inhibitor ( Inhibitor group) and control random sequences ( Control group) by liposome method.The levels of miR-186 were detected by QPCR, and proliferative ability was investigated by CCK-8. Transwell invasion and migration assays were used to detect the number of transmembrane cells. Double luciferase reporter assay was used to verify the interaction between miR-186 and ARAP2. Western blotting was used to detect the protein level of ARAP2. Results The miR-186 level in HN13 cells was higher than that in HOK cells (3. 945±0. 712 vs. 1. 049±0. 268, P＜0. 05). Compared with the Control group (1. 074±0. 192), the miR-186 level in Mimics group (4. 142±0. 348) increased, while that in Inhibitor group (0. 124± 0. 039) decreased with significant difference (P＜0. 05). Compared with the Control group, the proliferative activity of Mimics group increased at 24 and 48 h, while that of Inhibitor group decreased at 24 and 48 h (P＜0. 05).The numbers of perforating cells in Control group, Mimics group and Inhibitor group were 22. 9±3. 1, 71. 5±9. 6 and 15. 4±2. 4 in Transwell invasion experiment, and 51. 2±6. 1, 106. 7±12. 5 and 36. 8±5. 6 in migration experiment. Compared with Control group, the numbers of perforating cells in Mimics group were higher, while those in Inhibitor group were lower (P＜0. 05).MiR-186 could inhibit the relative luciferase activity in the wild type 3＇ untranslated region of ARAP2 ( P＜0. 05), but had no effect on the mutant type (P＞0. 05). Compared with the Control group (0. 436±0. 026), the level of ARAP2 in Mimics group (0. 024±0. 013) decreased, while that in Inhibitor group (0. 657±0. 019) in- creased (P＜0. 05). Conclusion The up-regulated expression of miR-186 in OSCC cells promotes the invasion and migration of OSCC cells. It may play an oncogenic role by targeting ARAP2 and become a target for diagnosis and new biological therapy of OSCC.