Mechanisms Of SCAMP1 Promoting Glioma Cells Malignant Biological Behaviors By Regulating MiR-499a-5p/LMX1A/NLRC5 Pathway

Objective:Glioma is the most prevalant malignant primary tumor of the central nervous system.Patients suffering from this desease meet a median of 15 months despite following surgery,chemotherapy,and radiotherapy,and the prognosis for patients with glioma is dismal and with high recurrence rate.Long non-coding RNAs(Inc RNAs)are involved in various genetic phenomena which is associated withtumorigenesis and malignant progression.The aim of this study is to investigate whether long non-coding RNASCAMP1 affected the malignant biological behavior of glioma cells and the possible molecular mechanism.We further aimed to explore whether SCAMP1 could promote LMX1 A and NLRC5 expression by negatively regulating miR-499a-5p Promote malignant biological behavior of GCs and promote malignant biological behavior of glioma cells.Methods:Human glioma cell line U87,U251,human normal astrocytes were cultured.Real-time PCR were used to detect the expression of SCAMP1,miR-499a-5p,LMX1 A and NLRC5,LMX1 A and NLRC5 was detected by Western blot in normal brain tissue,glioma tissue,human normal astrocytes and glioma cells.Cell proliferation was detected by Cell Counting Kit-8.Cell migration and invasion ability was measured using Transwell assay.Cell apoptosis was detected by flow cytometry.Inhibition of SCAMP1 and NLRC5 plasmids were transfected into the cells.Over-expression or inhibition of miR-499a-5p and LMX1 A plasmids were transfected into the cells.Real-time PCR was used to detect the m RNA expression of LMX1 A and NLRC5,and miR-499a-5p.The expression of LMXA1 A,NLRC5、clclin D1,c-Myc,β-catenin,MMP-7 was determined by Western blot.Regulation of miR-499a-5p targeting SCAMP1,regulation of miR-499a-5p targeting LMX1 A,regulation of miR-499a-5p targeting LMX1 A were evaluated by luciferase reporter assay and RNA immunoprecipitation.Results: SCAMP1 was up-regulated in GCs.Inhibition of SCAMP1 attenuated malignant biological.Mi R-499a-5p was downregulated glioma tissues and GCs.Over-expression of miR-499a-5p significantly restrained malignant biological behaviors of GCs.Mi R-499a-5p targeted SCAMP1 in a sequence-dependent manner.Knockdown of SCAMP1 impaired malignant biological behaviors of GCs by decreasing LMX1 A and NLRC5 expression,which were negatively mediated by miR-499a-5p.Over-expression of miR-499a-5p impairedmalignant biological behaviors of GCs by decreasing LMX1 A and NLRC5 expression,which regulated cleaced cyclin D1,c-Myc,β-catenin and MMP-7 expression.LMX1 A was up-regulated in GCs.LMX1 A targeted NLRC5 in a sequence-dependent manner.Knockdown of LMX1 A impaired malignant biological behaviors of glioma cells by decreasing NLRC5 expression,which regulated Wnt/ β-catenin pathways activitis.NLRC5 was up-regulated in GCs.Knockdown of NLRC5 impaired malignant biological behaviors of glioma cells by decreasing cyclin D1,c-Myc,β-catenin and MMP-7 expression,which regulated Wnt/ β-catenin pathways activitis.Conclusion:1.SCAMP1 was up-regulated in GCs,miR-499a-5p was down-regualted in glioma tissues and cells.Knockdown of SCAMP1 or over-expression of miR-499a-5p impaired malignant biological behaviors of GCs.2.Mi R-499a-5p targeted SCAMP1 in a sequence-dependent manner.LMX1 A targeted miR-499a-5p in a sequence-dependent manner.Knockdown of SCAMP1 impaired malignant biological behaviors of GCs by decreasing LMX1 A and NLRC5 expression,which were negatively mediated by miR-499a-5p.Over-expression of miR-499a-5p impairedmalignant biological behaviors of GCs by decreasing LMX1 A and NLRC5 expression,which regulated cleaced cyclin D1,c-Myc,β-catenin and MMP-7 expression,an Wnt/ β-catenin pathways activitis.3.LMX1 A and NLRC5 was up-regulated in GCs.Knockdown of LMX1 A impaired malignant biological behaviors of glioma cells by decreasing NLRC5 expression,which which regulated cleaced cyclin D1,c-Myc,β-catenin and MMP-7expression,and regulated Wnt/ β-catenin pathways activitis...