LncRNA SNHG15 Regulates Glioma Proliferation,invasion And Glycolysis Via Targeting MiR-138-5p/RhoC

Glioma is a primary brain tumor,which may originate from glial cells.It is characterized by high incidence of malignant glioma,high malignancy and high recurrence rate of malignant glioma.The molecular mechanism of the pathogenesis and progression of gliomas has been explored,which has potential diagnostic and therapeutic value.With the advent of high-throughput technology,a large number of long noncoding RNAs(lncRNAs)have been found in recent years.Some of them may be involved in the occurrence,malignant progression,recurrence and drug resistance of glioma,or of diagnostic and prognostic value.The purpose of this study was to investigate the expression and role of lncRNA SNHG15 in glioma.Part 1.Expression of lncRNA SNHG15 in gliomaObjective:To investigate the expression and clinical significance of lncRNA SNHG15 in gliomaMethods:1.A total of 90 pairs of glioma tissues and matched Peritumoral Brain Edema(PTBE)tissues were collected and the expression of SNHG15 was detected by qRT-PCR.2.The expression of SNHG15 in different stages(WHO standard)of glioma tissues was also analyzed.3.Kaplan Meier survival analysis was used to analyze the effect of SNHG15 on the survival of glioma patients.4.Pearson’s χ 2 was used to analyze the relationship between the expression of SNHG 15 and the clinical features of glioma patients.Results:1.The expression of SNHG 15 in glioma tissues was significantly higher than that in PTBE tissues.2.The expression of SNHG 15 in stage Ⅲ-Ⅳ glioma tissues was significantly higher than that in stage Ⅰ-Ⅱ glioma tissues.3.In glioma high expression of SNHG15 indicated a lower 5-year survival rate.4.The expression of SNHG15 is related with tumor size,peritumoral brain edema and WHO classification.Conclusion:LncRNA SNHG 15 was highly expressed in glioma tissues and its highly expression was associated with poor prognosis of glioma patients.Part 2.Knockdown of lncRNA SNHG15 inhibited biological activity of glioma cellsObjective:To investigate the effect of SNHG15 knockdown on the biological activity of glioma cellsMethods:1.Three SNHG15 interfering sequences were designed to knock down SNHG15 in U251 and ln229 cells,and the knockdown efficiency was detected by qRT-PCR.2.CCK-8 assay was used to detect cell proliferation.3.Cell apoptosis was detected by flow cytometry.4.Cell migration was detected by wound healing assay.5.Transwell assay was used to detect cell invasion.6.Glycolysis was detected by ELISA.Results:1.Knockdown of SNHG15 inhibited proliferation of glioma cells.2.Knockdown of SNHG15 promoted apoptosis of glioma cells.3.Knockdown of SNHG15 suppressed migration of glioma cells.4.Knockdown of SNHG15 decreased invasion of glioma cells.5.Knockdown of SNHG15 suppressed cell glycolysis of glioma.Conclusion:Knockdown of lncRNA SNHG15 inhibited biological activity of glioma cells in vitro.Part 3.LncRNA SNHG15 regulated cell proliferation,invasion and glycolysis via miR-138-5p/RhoC axis in gliomaObjective:To explore the molecular mechanism of the effect of SNHG15 on glioma in vitro.Methods:1.The Online biological tool miRDB was used to predict the target miRNA of SNHG15.2.Luciferase report assay was used to verify the prediction.3.RNA pull down assay was also used to verify the prediction.4.The expression of the predicted miRNA in glioma tissues was assessed using qRT-PCR.5.Spearman’s correlation analysis was used to calculate the correlation between miRNA expression and SNHG15 expression.6.The online biological tool TargetScan was used to predict the downstream mRNA of SNHG15.7.Luciferase report assay was used to verify the prediction.8.The expression of the predicted mRNA in glioma tissues was assessed using qRT-PCR.9.Spearman’s correlation analysis was used to calculate the correlation among mRNA,miRNA expression and SNHG15 expression.10.The expression of the predicted gene in glioma cell lines U251 and LN229 was detected using qRT-PCR and western blot.11.The axis was verified using a rescue experiment.Results:1.LncRNA SNHG15 targeted miR-138-5p.2.miR-138-5p expression was low in glioma tissues.3.In glioma tissues,the SNHG15 expression was reversely related with miR-138-5p expression.4.SNHG15/miR-138-5p targeted RhoC.5.RhoC expression was high in glioma tissues.6.In glioma tissues,the SNHG15 expression was positively related with RhoC expression,while the miR-138-5p expression was reversely related with it.7.Both RhoC knockdown and miR-138-5p up-regulation decreased the effect of SNHG15 high expression in glioma cells.Conclusion:LncRNA SNHG15 promotes cell proliferation,invasion and glycolysis via miR-138-5p/RhoC axis in gliomaPart 4.The in vivo antitumor effect of SNHG15 knockdown in gliomaObjective:To investigate the in vivo effect of SNHG15 knockdown in gliomaMethods:1.U251 cells were transfected with shNC or shSNHG15,the transfection efficiency was confirmed using qRT-PCR.2.The transfected U251 cells were injected subcutaneously into nude mice to establish glioma model.3.The tumor volume curve was drawn and the tumor was weighted.4.The expression of Ki-67 and RhoC was detected by immunohistochemistry.5.TUNEL assay was used to detect cell apoptosis.Results:1.Knockdown of SNHG15 inhibited glioma tumor growth in vivo.2.Knockdown of SNHG15 suppressed glioma tumor proliferation in vivo.3.Knockdown of SNHG15 promoted glioma tumor apoptosis in vivo.Conclusion:Knockdown of SNHG15 inhibited glioma proliferation and promoted glioma apoptosis in vivo.