A Study On MicroRNA-451a/YWHAZ Regulating Proliferation,Migration,Invasion And Apoptosis Of Lung Adenocarcinoma H1299Cells

Objective: To explore the effect of miR-451a/YWHAZ on the proliferation,migration,invasion and apoptosis of lung adenocarcinoma H1299 cells.Methods: 1.GEO database was used to screen out miRNAs of differential expression in NSCLC tissues and paracancerous tissues,and select miRNA-451 a with significant differences for research.2.qRT-PCR was used to detect the expression level of miR-451 a in BEAS-2B,A549,H1299 and H226 cells,and screen the miR-451 a low expression cells as the research object;To observe the effect of miR-451 a overexpression on the biological function of the experimental cells.3.The target of miR-451 a was predicted to be YWHAZ in the three bioinformatics databases of TargetScan,miRDB and miRanda;The dual-luciferase reporter assay system confirmed that YWHAZ and miR-451 a had binding sites.4.To observe the effect of knockdown YWHAZ on the biological function of the experimental cells.Results: 1.The expression of miRNA-451 a in 397 NSCLC tissues was lower than that in 151 paracancerous tissues in(P <0.05).2.The expression of miR-451 a in three NSCLC cell lines was significantly lower than that in human normal lung epithelial cells BEAS-2B,and the expression of human lung adenocarcinoma(1299)cells was the lowest;After overexpressing miR-451 a in H1299 cells,the results of clone formation assay showed that the number of clone formation in oe-miR451 a group(10.06±4.31/plate)was lower than that in oe-NC group(43.87±5.16/plate)(P <0.05);The results of wound healing assay showed that the healing rate in oe-miR451 a group(35.11%±5.23%)was significantly lower than that in oe-NC group(68.35%±6.61%)(P <0.05);The results of Transwell migration and invasion assay showed that the number of cell migration in the oe-miR451 a group(39.98±5.75/field)was significantly lower than that in the oe-NC group(119.69±12.29/field)(P <0.05),the number of cell invasions in the oe-miR451 a group(43.48±4.86/field)was significantly lower than that in the oe-NC group(103.97±7.11/field)(P <0.05);The results of flow cytometry showed that the apoptosis rate in the oe-miR451 a group(5.87%±1.04%)was significantly higher than that in the oe-NC group(1.10%±0.31%)(P <0.05).3.The target of miR-451 a was predicted to be YWHAZ in the three bioinformatics databases of TargetScan,miRDB and miRanda;The dual-luciferase reporter assay system suggested that YWHAZ was the target of miR-451 a.4.To observe the effect of knockdown YWHAZ on the biological function of the experimental cell lines;After knockdown YWHAZ in H1299 cells,the results of clone formation assay showed that the number of clone formation in si-YWHAZ group(16.42±4.43/plate)was ower than that in si-NC group(49.11±7.36/plate)(P <0.05);The results of wound healing assay showed that the healing rate in si-YWHAZ group(50.46%±4.21%)was significantly lower than that in si-NC group(86.92%±5.50%)(P <0.05);The results of Transwell migration and invasion assay showed that the number of cell migration in the si-YWHAZ group(64.62±6.55/field)was significantly lower than that in the si-NC group(134.96±10.31/field)(P <0.05),the number of cell invasions in the si-YWHAZ group(36.82±4.16/field)was significantly lower than that in the si-NC group(102.49±8.60/field)(P <0.05);The results of flow cytometry showed that the apoptosis rate in the si-YWHAZ group(12.07%±1.93%)was significantly higher than that in the si-NC group(2.99%±0.57%)(P <0.05).Conclusion: 1.MiR-451 a was low expression in NSCLC cell lines and overexpression of miR-451 a can inhibit the proliferation,migration and invasion of lung adenocarcinoma H1299 cells,and promote apoptosis of H1299 cells.2.MiR-451 a targeted YWHAZ to regulate the growth and function of lung adenocarcinoma cells,which may provide valuable targets and molecular markers for the diagnosis and treatment of lung adenocarcinoma.