Inhibition Of Proliferation And Migration Of Glioma Cells By MiRNA-1224-5p Via LNC00665

Recent studies have shown that LncRNA,an organizational framework of subcellular structure,regulates protein activity as the kind of RNA and becomes a key regulatory molecule in cells.LncRNA may also be involved in the epigenetic regulation of target genes,thereby inhibiting the transcription of target genes.LncRNA is closely related to tumor development,and may be an important biomarker and target for tumor diagnosis,treatment and prognosis.At present,the research group found Lnc00665 is a proto-oncogene,which could promote the proliferation of glioma cells,However,the regulatory effect of miRNA on Lnc00665 is still not very clear,So the study will explore the regulatory effect of miRNA on Lnc00665 from the perspective of mir-1224-5p inhibitor.The research confirmed that Lnc00665 was directly regulated by mirna-1224-5p by using qRT-PCR,double luciferase reporting system detection and mutation experiments based on bioinformatics analysis.In the previous work,glioma cells were transfected with miRNA-1224-5p instantaneously,and the over-expression inhibited the proliferation,migration and invasion of glioma cells.Therefore,we investigated the mechanism of proliferation and migration of glioma cells after transfection with mirna-1224-5p inhibitor.In order to further investigate the effect of miRNA-1224-5p inhibitor on the proliferation,migration and invasion of U251 and U87 cells,we transfected miRNA-1224-5p into glioma U251 and U87 cells.MTT proliferation curve and cloning analysis showed that when miRNA-1224-5p inhibitor was transfected into glioma cells,the proliferation ability of U251 and U87 cells was significantly increased.According to cell cycle experiments,miRNA-1224-5p inhibitor was observed to stall in G2 phase of cell cycle after transfection.MiRNA-1224-5p inhibitor was inhibitor of G2 phase of cell cycle after transfection.Western blot experiments showed that the expression level of cyclinD1 was significantly increased after transfection with miRNA-1224-5p inhibitor.After Transwell assay,we found that the migration and invasion of U251 and U87 cells were significantly increased after the transfection of miRNA-1224-5p inhibitor.These results demonstrated that miRNA-1224-5p inhibitor promoted the proliferation,migration and invasion of U251 and U87 cells aftertransfection.Meanwhile,nude mice overexpressed Lnc00665 were tested for tumor formation in vivo.Female BALB/ c-nu nude mice of 4 weeks old were selected,and U251 cell suspension with overexpressed Lnc00665 was subcutaneously injected into the right back.The mass of the tumor was dissected and weighed.The expression level of Lnc00665 in tumorigenic tissues of nude mice was detected by real-time quantitative PCR,and the expression level of Lnc00665 was significantly increased.The expression of proliferation marker PCNA was detected by immunohistochemistry,and the positive number of PCNA cells in glioma tissues with Lnc00665 overexpression was significantly increased,further verifying that Lnc00665 overexpression promotes glioma cell proliferation in vivo.The above results demonstrate that Lnc00665 is regulated by miRNA-1224-5p,which affects the proliferation,migration and invasion of glioma cells.This study will give assistance to the diagnosis and treatment of glioma cancer.