The inhibitory effect of MIR-154-3p on metastasis of non-small cell lung cancer cell lines / 肿瘤

ABSTRACT

Objective: To investigate the expression of microRNA-154-3p (miR-154-3p) in non-small cell lung cancer (NSCLC) tissues and cell lines, and explore the role of miR-154-3p in metastasis process of NSCLC. Methods: The expression levels of miR-154-3p in NSCLC tissues and the corresponding paracarcinoma tissues, metastatic and non-metastatic NSCLC tissues, NSCLC cell lines (A549, H292, H1975, 95D and SPC-A-1) and human embryo lung WI38 cells were detected by realtime fluorogenic quantitative-PCR (RFQ-PCR). The miR-154-3p-negative control (miR-154-3p-NC, as a negative control) or miR-154-3p-mimics was transiently transfected into A549 and H292 cells by liposome. The effects of miR-154-3p on migration and invasion abilities of NSCLC A549 and H292 cells were determined by wound healing assay and Transwell invasion assay. The expressions of epithelial-mesenchymal transition (EMT)-related proteins E-cadherin, zonula occludens-1 (ZO-1) (epithelial marker), N-cadherin and vimentin (mesenchymal marker) induced by miR-154-3p overexpression were detected by Western blotting. The bioinformatic softwares were used to predict the potential targets of miR-1 54-3p, and the interested target genes were further validated by Dual-Luciferase Reporter Assay and Western blotting. Results: The expression levels of miR-154-3p were significantly lower in NSCLC tissues (P < 0.001), metastatic NSCLC tissues (P < 0.05) and NSCLC cell lines (P < 0.01) than those in their corresponding controls. Overexpression of miR-154-3p attenuated the migration and invasion abilities of both A549 and H292 cells (P < 0.05). The expressions of E-cadherin and ZO-1 proteins were up-regulated in A549 and H292 cells (all P < 0.05), whereas the vimentin expression in A549 cells and N-cadherin expression in H292 cells were down-regulated (both P < 0.05). Dual-Luciferase Reporter Assay and Western blotting results revealed that miR-154-3p could negatively regulate the expression of BMI1. Conclusion: MiR-154-3p is lowly expressed in NSCLC tissues and cell lines. The overexpression of miR-154-3p can inhibit the migration and invasion abilities of NSCLC cells and block the EMT progression. BMI1 may be one target gene of miR-154-3p.