CircFOXK2 regulates the proliferation,migration,and invasion of non-small cell lung cancer through the miR-588/FBXW5 axis
Objective To investigate the impacts of circFOXK2 on the proliferation,migration,and invasion of non-small cell lung cancer(NSCLC)through the miR-588/FBXW5 axis.Methods Real-time fluorescence quantitative PCR(qRT-PCR)and Western blot were applied to detect the expression levels of circFOXK2,miR-588,and FBXW5 in the NSCLC cell line A549 and the normal human bronchial epithelial(NHBE)cell line.A549 cells were treated with blank control,or transfected with si-NC,si-circFOXK2,si-circFOXK2+inhibitor NC,and si-circFOXK2+miR-588 inhibitor.The relationship among CircFOXK2,miR-588 and FBXW5 was tested by the dual-luciferase reporter assay.Western blot was applied to detect the protein levels of FBXW5,E-cadherin,N-cadherin and Vimentin in A549 cells.Cell proliferation and apoptosis in A549 cells were detected by CCK-8 assay and flow cytometry,respectively.Invasive and migratory cell numbers were measured by Transwell assay.Results Compared with those co-transfected with miR-NC + WT-FOXK2,relative luciferase activity was significantly lower in A549 cells co-transfected with miR-588 mimic+WT-FOXK2(P<0.05).There was no significant difference in the relative luciferase activity between A549 cells co-transfected with miR-588 mimic+WT-FOXK2 and miR-588 mimic+MUT-FOXK2(P>0.05).Relative luciferase activity was significantly lower in A549 cells co-transfected with miR-588 mimic+WT-FBXW5 than those co-transfected with miR-NC+WT-FBXW5.No significant difference in the relative luciferase activity was detected between A549 cells co-transfected with miR-588 mimic+MUT-FBXW5 and miR-NC+MUT-FBXW5(P>0.05).Compared with those of NHBE cells,circFOXK2 and FBXW5 were significantly upregulated(P<0.05),and miR-588 was significantly downregulated in A549 cells(P<0.05).Transfection of si-circFOXK2 significantly downregulated circFOXK2,FBXW5,N-cadherin and Vimentin,reduced optical density at 450 nm wavelength(OD450)and migratory and invasive cell numbers(P<0.05),and upregulated miR-588 and E-cadherin and increased apoptotic rate in A549 cells than those treated with blank control or transfected with si-NC(P<0.05).Knockdown of miR-588 significantly reversed the regulatory effects of silenced circFOXK2 on favoring the proliferation,migration and invasion of A459 cells.CircFOXK2 targeted and regulated the miR-588/FBXW5 axis.Conclusion Silencing circFOXK2 downregulates FBXW5 by upregulating miR-588,thereby affecting the proliferation,migration,and invasion of NSCLC cells.
CircFOXK2MiR-588FBXW5non small cell lung cancerproliferationmigrationinvasion
NETL
NSTL
万方数据
维普
