Effect of microRNA-296-3p targeting Notch2 on migration and invasion of non-small cell lung cancer
Objective To investigate the impact of microRNA(miR)-296-3p targeting Notch2 on the migration and invasion of non-small cell lung cancer(NSCLC).Methods Tissue specimens from 50 cases of lung adenocarcinoma,both cancer and adjacent tissues,were collected at the Department of Thoracic Surgery,Nantong First People's Hospital,during April 2022 to March 2023,including 30 males and 20 females.No lymph nodes and distant metastases were found in postoperative pathology.The expres-sion of miR-296-3p in cancer and adjacent tissues was analyzed using fluorescence quantitative real-time polymerase chain reaction(qRT-PCR).Lentivirus was utilized to engineer A549 cells overexpressing miR-296-3p,labeled as A549/miR-296-3p,with A549/miRNA cells as the control.The Transwell assay was used to explore the impact of miR-296-3p overexpression on the migratory and invasive capabilities of A549 cells.Concurrently,Western blotting analysis was performed to analyze the expression levels of Notch2 in the A549 cells across different groups.Dual luciferase reporter assay was used to analyze whether Notch2 was a target gene of miR-296-3p.T-test was used for the comparison of data between groups.Results The qRT-PCR results suggest that the expression level of miR-296-3p in the adjacent tissues of non-small cell lung cancer(4.215±1.600)was significantly higher than that in NSCLC(1.931±0.700)(n=40,t=9.539,P＜0.01).The expression of miR-296-3p was significantly elevated in bronchial epi-thelial cells BEAS-2B(1.018±0.075)compared to A549 cells(0.581±0.014)(n=3,t=9.842,P＜0.01).The expression level of miR-296-3p in A549/miRNA cells(0.846±0.047)was substantially low-er than in the A549/miR-296-3p cell group(2.266±0.073)with the difference being statistically signifi-cant(n=3,t=28.200,P＜0.01).The results of the Transwell migration assay revealed that the number of migrating A549 cells per field of view in the A549/miRNA control group(242.560±7.550)was signifi-cantly greater than in the A549/miR-296-3p group(86.120±5.570)(n=3,t=28.800,P＜0.01).The number of invasive A549 cells per field of view in the A549/miRNA control group(114.700±6.510)was significantly greater than in the A549/miR-296-3p group(48.000±3.610)(n=3,t=15.520,P＜0.01).Bioinformatics results suggested that Notch2 might be a target gene of miR-296-3p,which was con-firmed through a dual-luciferase reporter gene assay.Conclusion MiR-296-3p significantly influences the migration and invasion of NSCLC by regulating the expression of Notch2.
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