MicroRNA-876-5p Targets FOXM1 To Regulate Proliferation,Apoptosis,Invasion And Migration Of The Glioblastoma

Objective:(1)To observe the expression of miR-876-5p and FOXM1 in human GBM tissues and GBM cell lines in vitro.(2)To observe the regulation of proliferation,apoptosis,invasion and migration of GBM by miR-876-5p targeting FOXM1 in vitro and proliferation of GBM in vivo.Methods:(1)qRT-PCR was used to detect the expression level of miR-876-5p nucleic acid in GBM cell lines U138,U251,T98,LN229,clinical samples and normal brain cell lines NHAs.miR-876-5p mimics(miRNA-876-5p mimics)and miR-876-5p blank control(miR-NC)were transfected into GBM cell lines U251 and T98 by Lipofectamine 2000 method,and transfection efficiency was detected by qRT-pCR and Western Blot.Proliferation,apoptosis,migration and invasion of GBM cells transfected with miR-876-5p mimics and miR-NC were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively;(2)Online miRNA prediction software was used to predict the putative targets of miR-876-5p as FOXM1.The targeting association between miR-876-5p and FOXM1 was verified by the dual luciferase reporter system.The nucleic acid and protein expression level of FOXM1 in clinical GBM samples was detected by qRTPCR and Western Blot.Spearman's correlation regression analysis was used to analyze the correlation between the expression trend of miR-876-5p and FOXM1.miR-876-5p mimics(miR-876-5p mimics)and miR-876-5p blank control(miR-NC)were transfected into GBM cell lines U251 and T98 by Lipofectamine 2000 method.qRT-PCR and Western Blot were used to detect the expression of FOXM1 at transcription and translation levels regulated by miR-876-5p.FOXM1 RNA interference vector(FOXM1 siRNA)and negative control(NC siRNA)were constructed.Proliferation,apoptosis,migration and invasion of GBM cells transfected with FOXM1 siRNA and NC siRNA were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively;(3)Lipofectamine 2000 was used to transfect miR-876-5p mimics+pCMV,miR-876-5p mimics+pCMV-FOXM1 and miR-NC into U251 and T98 cell lines,and the transfection efficiency was detected by qRT-PCR and Western Blot.Proliferation,apoptosis,migration and invasion of GBM cells transfected with miR-876-5p mimics+pCMV,miR-876-5p mimics+pCMVFOXM1 and miR-NC were detected by CCK-8 assay,flow cytometry and Transwell assay,respectively;To establish a nude mouse model,U251 and T98 cells transfected with miR-876-5p mimics+pCMV,miR-876-5p mimics+pCMV-FOXM1 and miR-NC were inoculated subcutaneous in the neck and back of nude mice,respectively,and the differences in tumor volume and weight were observed.Results:(1)MiR-876-5p was significantly lower in human GMB specimens and GBM cell lines than in normal controls.After transfection with miR-876-5p mimics,the expression of miR-876-5p in GBM cells was increased.After transfection with miR-876-5p mimics,the proliferation,apoptosis,migration and invasion of GBM cells decreased.(2)FOXM1 was predicted to be the main target of miR-876-5p.There was a targeting association between miR-876-5p and FOXM1.The expression of FOXM1 in GMB samples was increased,which was contrary to the expression trend of miR-876-5p.There was a negative regulatory trend.After miR-876-5p was up-regulated,FOXM1 nucleic acid and protein expression levels were significantly decreased.After transfection with FOXM1 siRNA,the proliferation,apoptosis,migration and invasion of GBM cells decreased.(3)After transfection with miR-876-5p mimics+pCMV,the protein expression level of FOXM1 in GBM cells decreased,and transfection with miR-876-5p mimics+pCMVFOXM1 restored the protein expression level of FOXM1.After transfection with miR-876-5p mimics+pCMV,the proliferation,apoptosis,migration and invasion of GBM cells decreased.After transfection with miR-876-5p mimics+pCMV,the proliferation,migration and invasion of GBM cells were restored,and apoptosis was decreased.Tumor volume and weight increased significantly in nude mice transfected with miR-876-5p mimics+pCMV GBM.Conclusion:(1)miR-876-5p expression was decreased in human GBM tissue samples and in vitro GBM cells,suggesting that miR-876-5p expression was related to malignant biological behavior of GBM.miR-876-5p can inhibit the proliferation,migration and invasion of GBM cells and promote apoptosis.(2)Increased expression of FOXM1 nucleic acid and protein in human GBM tissue samples was associated with the malignant biological behavior of GBM.FOXM1 promotes proliferation,migration and invasion of GBM cells,and inhibits apoptosis.(3)FOXM1 is directly regulated by the upstream gene miR-876-5p,and the regulation between miR-876-5p and FOXM1 plays an important role in the pathogenesis of GBM.