Effects of ebv-miR-BART7 on tumorigenicity, metastasis, and TRAIL sensitivity of non-small cell lung cancer

Abstract

Objective: To investigate how the Epstein-Barr virus (EBV) encoded microRNA BART7 (miR-BART7) affects tumorigenicity, metastasis, and TRAIL sensitivity of non-small cell lung cancer (NSCLC). Methods: Real time-polymerase chain reaction was performed to detect miR-BART7 expression in NSCLC cell lines. A549 and Calu-1 cells transfected with miR-BART7 inhibitors/mimics were used to do the in-vitro experiments, including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Annexin V-fluorescein isothiocyanate/PI, wound-healing, transwell, clonogenic assays, Western blot analysis, and anchorage-independent growth assay. Additionally, mice were used to inject A549 cells infected with miR-BART7 inhibitors to observe the tumorigenicity and metastasis of NSCLC. Results: TRAIL-resistant NSCLC cell lines (H460R, A549, Calu-1, and H1299) exhibited higher miR-BART7 rather than sensitive H460 and H292 cells. After transfected with miR-BART7 inhibitors, we observed an inhibition in proliferation, migration, invasion, and colony formation, but an enhancement in apoptosis as well as expressions of caspase-3 and caspase-8 in A549 and Calu-1 cells. Besides, TRAIL elevated the migration, invasion, and anchorage-independent growth of A549 cells, which was reversed by silencing DR4 and DR5 (siDRs). However, miR-BART7 inhibitors could reduce migration, invasion, and transformation potential of TRAIL treated A549 cells. Moreover, the expression of transforming growth factor-beta 1 (TGFβ1) could be decreased by miR-BART7 inhibitors with or without TRAIL treatment. Moreover, the tumor growth, epithelial-to-mesenchymal transition, and metastasis was suppressed and tumor-free survival was extended after injection of A549-miR-BART7 inhibitors. Conclusion: Inhibition of miR-BART7 exerted inhibitory effects on cell proliferation, migration, invasion, and colony formation, consequently facilitating cell apoptosis and raising TRAIL sensitivity, providing a new therapeutic target in NSCLC.