| Backgroundglioma is the most common and fatal primary tumor of the central nervous system.glioblastoma(GBM)accounts for 50%of all gliomas,and it is one of the most malignant diseases with poor prognosis.In recent years,with the development of high-throughput sequencing technology,studies on the mechanism of long non-coding RNA(lncRNA)in the occurrence and development of glioma have been found.Many studies have shown that lncRNA can affect the expression of its downstream target mRNA by sponge adsorption of miRNA,and form a competing endogenous RNA(ceRNA)regulatory network to regulate tumor progression.Therefore,clarifying the regulatory mechanism of ceRNA network mediated by lncRNA in glioma will provide new strategies for early diagnosis and target therapy of the disease.LINC01579,a lncRNA differentially expressed in GBM,was screened by lncRNA expression profile chip in glioblastoma tissues,and the ceRNA regulatory network mediated by LINC01579 was found to affect the proliferation,invasion and migration of glioma.This study further clarified the molecular mechanism of glioma development,and the ceRNA network involved in LINC01579 may be used as a target for early diagnosis and molecular treatment of glioma.MethodsBrain tissue microarray was used to screen out the differentially expressed lncRNA LINC01579.GEPIA database was used to analyze the expression of LINC01579 in glioma,and qRT-PCR was used to verify the expression of LINC01579 in glioma tissues and cells.The LINC01579 overexpression and knockdown cell systems were constructed,and the effects of LINC01579 on the proliferation,invasion and migration of glioma cells were verified by CCK-8 assay and Transwell assay.The downstream miRNA and downstream target mRNA of miRNA in LINC01579-mediated ceRNA network were identified by nuclear and cytoplasmic separation,dual luciferase gene reporter,RNA immunoprecipitation,and Western blot.Rescue experiments were designed to verify that LINC01579 promoted the proliferation,invasion and migration of glioma through the ceRNA network mechanism.Result1.The expression of LINC01579 was significantly increased in gliomasThe expression of LINC01579 was significantly up-regulated in glioblastomas.GEPIA database analysis showed that LINC01579 expression was significantly increased in glioblastomas with tissue specificity.2.LINC01579 promoted the malignant biological behavior of glioma cellsThe proliferation,invasion and migration of glioma cells with overexpression of LINC01579 were significantly higher than those of the control group.Knockdown of LINC01579 inhibited the proliferation,invasion and migration of glioma cells.3.LINC01579 served as sponge for miR-23b-3pEncori database analysis of miRNA with binding sites to LINC01579 showed that qRT-PCR showed that the expression of miR-23b-3p increased most significantly when LINC01579 was knocked out.Dual luciferase analysis and RIP experiment showed that LINC01579 could sponge adsorb miR-23b-3p.Pearson correlation analysis showed that LINC01579 was negatively correlated with miR-23b-3p.4.miR-23b-3p partially reversed the effect of LINC01579 on gliomaIn the rescue experiment,overexpression or knockdown of miR-23b-3p partially reversed the effects of LINC01579 on the proliferation,invasion and migration of glioma cells.5.NFE2L2 is the target mRNA of miR-23b-3pThe Encori database predicted that miR-23b-3p had a binding site with the 3’untranslated regions(UTR)of NFE2L2 mRNA.Dual luciferase reporter assay confirmed that NFE2L2 was the downstream target of miR-23b-3p.western blot assay showed that overexpression of miR-23b-3p decreased the expression of NFE2L2 protein,while knockdown of miR-23b-3p increased the expression of NFE2L2 protein.6.LINC01579 promotes the malignant progression of glioma cells through miR-23b-3p/NFE2L2 regulatory axisCCK-8 and Transwell experiments showed that NFE2L2 promoted the proliferation,invasion and migration of glioma cells.Overexpression of LINC01579 increased the protein expression of NFE2L2,while knockdown of LINC01579 decreased the protein expression of NFE2L2.Western blot assay showed that overexpression of miR-23b-3p or inhibition of miR-23b-3p partially reversed the effect of LINC01579 on the protein expression level of NFE2L2.Pearson correlation analysis of GEPIA database showed that the expression of LINC01579 was positively correlated with NFE2L2 in GBM..ConclusionThe results of this study confirmed that LINC01579 expression is significantly increased in glioblastoma with certain tissue specificity,and LINC01579 acts as a competing endogenous RNA to regulate the expression of NFE2L2 by sponge adsorption of miR-23b-3p,thereby promoting the malignant progression of glioblastoma cells. |
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