| Objective: To investigate the expression of LINC01235 in glioma and its effect on invasion and migration of glioma cells.Methods: Expression level data and clinical information of glioma tissue samples were obtained from The Cancer Genome Atlas(TCGA)database.1.Gene Expression Profiling Interactive Analysis(GEPIA)was used to investigate the expression of LINC01235 in normal brain tissues and gliomas of different grades.2.The relationship between the expression level of LINC01235 and the survival of patients was analyzed by survival.3.The effect of LINC01235 on the prognosis of glioma patients was analyzed by univariate and multivariate COX analysis.4.The accuracy of LINC01235 in predicting patients’ survival was evaluated by constructing a column diagram.5.Gene set enrichment analysis(GSEA)was used to predict the potential biological mechanism of LINC01235.6.The regulatory relationship between LINC01235 and related genes was analyzed by co-expression.7.Gene ontology(GO)analysis and pathway enrichment(KEGG)analysis were performed using the differentially expressed genes of LINC01235.8.To evaluate the difference between high and low expression of LINC01235 in tumor microenvironment,tumor mutation and immune-related aspects.9.The p RRophetic package in R language was further used to predict drug sensitivity.10.The expression of LINC01235 in glioma tissues and cells was verified by q RT-PCR.11.Transwell and cell scratch assay were used to detect the effects of LINC01235 on glioma cell invasion and migration.Research results: 1.The expression level of LINC01235 in glioma is high and the survival rate is low(1)The expression level of LINC01235 in low-grade glioma(LGG)and glioblastoma(GBM)was significantly up-regulated compared with that in normal tissues,with significant statistical significance(p < 0.05).(2)According to the survival analysis of GEPIA,LINC01235 was divided into high and low according to the median expression.In both groups,the survival rate of LINC01235 was lower in the high expression group than in the low expression group,suggesting poor prognosis.2.Evaluation of prognostic significance and prediction accuracy of LINC01235(1)Univariate COX analysis showed that LINC01235 was significantly correlated with the prognosis of patients.(2)Multivariate COX analysis showed that LINC01235 could be used as an independent prognostic factor in glioma patients.(3)According to ROC curve,the 1-year,3-year and 5-year survival rates of LINC01235 expression were 0.61,0.64 and 0.71,respectively.(4)According to the column chart and calibration curve,LINC01235 can predict the survival of patients with high accuracy.3.GSEA enrichment analysis of LINC01235 The results of GSEA showed that the peroxisome was significantly active in the high expression group of LINC01235,while the ERBB signaling pathway,WNT signaling pathway,MTOR signaling pathway,phosphatidylinositol signaling system and type 2 diabetes mellitus were significantly active in the low expression group.4.Co-expression analysis of LINC01235 Co-expression analysis showed that LINC01235 was positively correlated with PON2,LINC01338,PACRG,LINC01778 and HCG22,and negatively correlated with TOMM20,MMS19,SFXN3,ZFYVE27 and RIPOR1.5.Differential analysis of LINC01235 expression According to the median expression level of LINC01235,Lin C01235 was divided into high and low expression groups,and the related differential genes were found by difference analysis(P < 0.05).6.GO and KEGG enrichment analysis of differential genes(1)GO analysis showed that it was related to ciliary movement,transmembrane transport and neurotransmitters.(2)KEGG analysis showed that it was related to neural active ligand-receptor interaction,c AMP signaling pathway and calcium signaling pathway.7.Difference analysis of tumor microenvironment The results showed that LINC01235 had significant differences in the tumor microenvironment,and was significantly up-regulated in the high expression group.8.Immune cell and immune checkpoint analysis of LINC01235(1)Analysis of immune cell differences showed that the expression level of LINC01235 was significantly different in T cells and macrophages.(2)Immune cell correlation analysis showed that LINC01235 was positively correlated with resting mast cells and negatively correlated with activated mast cells.(3)The correlation analysis of immune checkpoint showed that LINC01235 was correlated with 14 of the 47 immune checkpoint related genes,among which,LINC01235 was negatively regulated with TNFSF9 and positively regulated with the other genes.9.Correlation between LINC01235 expression and tumor somatic cell genome(1)Tumor mutation load analysis showed that the expression of LINC01235 was positively regulated with tumor mutation load.(2)Immunotherapy analysis showed that there was no significant difference between the two groups receiving anti-PD1 and/or anti-ct LA4 treatment with high or low expression of LINC01235.10.Drug sensitivity analysis of LINC01235 Drug sensitivity analysis showed that the IC50 values of loskovidine,temozolomide,cipropylamine,phenylguanidine and lapatinib in the high expression group of LINC01235 were lower than those in the low expression group.11.In vitro experiments confirmed the high expression of LINC01235 in brain glioma By q RT-PCR assay,LINC01235 was highly expressed in glioma tissues,which was consistent with the predicted results of bioinformatics analysis.12.Expression level of LINC01235 in glioma cell lines U87 and U251 LINC01235 was highly expressed in U87 glioma cell line and relatively low in U251 glioma cell line.13.Knockdown of LINC01235 inhibited the migration and invasion ability of U87 cells(1)Cell scratch test,by observing the migration of cells at 0 hours and 12 hours,it was found that the migration ability decreased significantly.(2)Transwell assay showed that the cell invasion ability decreased significantly after LINC01235 knockdown.Conclusions: This study shows that the expression level of LINC01235 is negatively correlated with the prognosis of glioma patients.LINC01235 can promote glioma cell migration and invasion in vitro.In addition,LINC01235 may serve as a novel biomarker for diagnosis,treatment and prognosis assessment of glioma. |
