Effect of microRNA-520e on invasion, migration and JAK1／STAT3 signaling pathway of non-small cell lung cancer cells
YAN Chengyi
YANG Jicheng
YUAN Yuexi
Abstract：Objective To investigate the effect of microRNA-520e ( miR-520e) on invasion, migration and Janus kinase 1 (JAK1)／signal transducer and activator of transcription 3 ( STAT3) signaling pathway of non-small cell lung cancer ( NSCLC) cells. Methods The miR-520e levels of normal pulmonary epithelial cell BEAS-2B and NSCLC cell lines H1299, H1975, H1650 and A549 were detected by real-time quantitative PCR (QPCR). A549 cells were cultured in vitro and transfected with mimics ( overexpression group) and negative control sequence (NC group) by liposome, respectively. Only cells exposed to liposome were chosen as the control group. The levels of miR-520e, zinc finger and BTB domain containing 7A (ZBTB7A), proliferation activity, scratch healing rate and number of membrane penetrating cells were detected by QPCR, cell counting kit (CCK)-8, scratch test and Transwell test, respective-ly. The levels of matrix metalloproteinase ( MMP)-9, phosphorylated JAK1 (p-JAK1) and phosphorylated STAT3 (p-STAT3) were detected by Western blotting. The double luciferase reporter gene system combined with luciferase reporter plasmid psiCHECK-2 were applied to verify the targeting relationship between miR-520e and ZBTB7A. Results The miR-520e levels in NSCLC cells was lower than that in normal cells, and A549 cells was the lowest ( P＜0. 05). The miR-520e level was 19. 562± 1. 448 in the overexpression group, higher than 1. 002±0. 057 in control group and 1. 053±0. 082 in NC group, and ZBTB7A level was 0. 173±0. 026, lower than 1. 109±0. 091 in control group and 1. 076±0. 136 in NC group (P＜0. 05). Compared with control group and NC group, the prolifera-tion activity at 36 and 48 h after transfection and levels of MMP-9, p-JAK1 and p-STAT3 at 48 h after transfection were lower in the overexpression group (P＜0. 05). The scratch healing rate mimics and the number of membrane penetrating cells in the overexpression group were (23. 618± 3. 684)% and 51. 644 ± 7. 069, lower than (57. 089 ± 4. 569)% and 152. 269 ± 12. 833 in control group and (59. 267±5. 671)% and 149. 075± 11. 239 of NC group ( P＜0. 05). The decreased relative fluorescence intensity were observed in cells co-transfected with miR-520e mimics and the psiCHECK-2 vector carrying wild-type ZBTB7A 3' untranslated region ( P＜0. 05). Conclusion MiR-520e is down regulated in NSCLC cells, and it may inhibit the proliferation, invasion and migration of NSCLC cells possibly by targeting ZBTB7A to reduce activity of JAK1／STAT3 signal pathway.
Keywords：Non-small cell lung cancerMicroRNA-520eInvasion and migrationZinc finger and BTB domain 7A
Publication Date：2019-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 978-983 )