Targeted regulation of microRNA-488 on HMGN5 expression and its effect on invasion and migration of lung cancer cells

SUN Wei; DING Jing

doi:10.3969/j.issn.1009-0460.2019.11.003

Abstract：Objective To investigate the effect of microRNA-488 (miR-488) on the targeting regulation of high-mobility group nucleosome binding domain 5 (HMGN5) and the invasion and migration of non-small cell lung cancer (NSCLC) A549 cells. Methods Real-time quantitative PCR (QPCR) was used to detect the miR-488 level in normal pulmonary epithelial cell line (BEAS-2B) and A549 cells. A549 cells were transfected with miR-488 mimics (Overexpression group) or negative control sequence (NC group), and the un-transfected cells were used as Control group. The proliferation, invasion and migration ability of A549 cells were measured by MTT, Tran-swell and scratch healing. Levels of HMGN5, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by QPCR and Western blot-ting. The double luciferase reporter gene experiment was applied to verify the targeted regulation of miR-488 on HMGN5. Results The miR-488 level of A549 cells was 0. 256±0. 028, lower than 1. 019±0. 124 of BEAS-2B cells (P＜0. 05). The miR-488 level in Overexpres-sion group was 11. 229±2. 164, higher than 1. 051±0. 162 in Control group and 1. 029±0. 116 in NC group (P＜0. 05). Compared with the Control group and NC group, the cell proliferation ability of the Overexpression group decreased after 48 and 72 h-transfection (P＜0. 05). The scratch healing rate and number of membrane piercing cells in the Overexpression group were (25. 53±3. 75)% and 79. 66±9. 23, lower than (54. 34±5. 65)% and 165. 22±11. 53 in Control group and (58. 56±4. 76)% and 172. 34±14. 67 of NC group ( P＜0. 05). MiR-488 could reduce the luciferase activity of the wild type of 3’ untranslated region of HMGN5 gene (P＜0. 05), but had no effect on the mutant type of 3’ untranslated region (P＞0. 05). Compared with other two groups, the levels of HMGN5, MMP-2 and MMP-9 in Overexpression group decreased (P＜0. 05). No significant difference was observed on the above indices between Control group and NC group (P＞0. 05). Conclusion MiR-488 is lowly expressed in NSCLC cells. Up-regulation of miR-488 can inhibit proliferation, migration and invasion, and play a role similar to tumor suppressor gene. It may be related to targeted regulation of HMGN5 and inhibition of MMP-2／MMP-9 pathway, which has certain reference value in NSCLC targeting therapy.