Targeted regulation of microRNA?592 on JAK1 expression and its effect on invasion and migration of non?small cell lung cancer cells

WEI Haixia; SONG Yongna; TIAN Wenxian

doi:10.3969/j.issn.1009-0460.2019.08.001

Abstract：Objective To investigate the targeted regulation of microRNA-592 (miR-592) on Janus kinase 1 (JAK1) and its effect on migration and invasion of non-small cell lung cancer (NSCLC) cells. Methods The levels of miR-592 in NSCLC cells A549 and human normal lung epithelial cells BEAS-2B were detected by real-time quantitative PCR (QPCR). Then, A549 cells were trans-fected with miR-592 mimics (Overexpression group) and negative control ( NC group) by Lipofectamine 2000. The level of miR-592 was detected by QPCR after transfection. Scratch healing rate and number of penetrating cells were measured by scratch test and Tran-swell chamber test to evaluate migration and invasion ability, respectively. Double luciferase reporter gene assay was used to verify the targeting regulation of miR-592 on JAK1. Western blotting was used to detect the protein levels of JAK1, phospho-JAK1 ( p-JAK1), phospho-signal transducer and activator of transcription 3 (p-STAT3) and matrix metalloproteinase-9 (MMP-9). Results The results of QPCR showed that the level of miR-592 in A549 cells was 0. 312±0. 097, lower than 1. 062±0. 136 in BEAS-2B cells (P＜0. 05). The relative expression of miR-592 in A549 cells of Overexpression group was 6. 647±1. 847, higher than 1. 054±0. 168 in NC group (P＜0. 05). The healing rate and the number of perforating cells were (27. 45±2. 51)% and 105. 2±15. 8 in the Overexpression group, lower than (54. 45±3. 06)% and 312. 4±19. 2 in the NC group with significant difference (P＜0. 05). MiR-592 mimics inhibited the relative luciferase activity of wild-type 3′-untranslated region of JAK1 in A549 cells, but had no influence on the mutant. Compared with NC group, the levels of JAK, p-JAK1, p-STAT3 and MMP-9 protein in Overexpression group were significantly decreased ( P＜0. 05). Conclusion MiR-592 negatively regulates the invasion and migration of NSCLC cells, which may be related to the inhibition of JAK／STAT3 signaling pathway, suggesting that miR-592 plays an anti-cancer role and a target for treatment of NSCLC.