| Objective:1.To analyze the differential expression of circRNAs between glioblastoma and normal brain tissue;2.To compare the expression of hsa circ 0008278 in glioblastoma and normal brain tissue,and to analyze its correlation with clinical features and prognosis of glioma patients;3.To interfere with the expression of hsacirc0008278 in glioma cell lines:U87,U251 and LN229 and observe its effect on glioma growth.Method:1.Six cases of glioblastoma tissue and 6 normal brain tissues were sequenced by RNA-seq.Differentially expressed circRNAs were analyzed with the fold change ?2 times and p?0.05.GO analysis and KEGG pathway analysis were performed on host genes of differentially expressed circRNAs.The differentially expressed circRNAs were randomly selected and detected by RT-qPCR in 6 normal brain tissues and 45 glioma tissues to verify the sequencing results.2.RT-qPCR method was selected to detect expression level of hsa circ 0008278 in 6 normal brain tissues and 45 glioma tissues.The corresponding clinical data of glioma patients were collected,and the correlation between the expression level of hsacirc0008278 and the clinical features and prognosis of glioma patients was analyzed by SPSS23.3.hsa circ 0008278 knockdown and over expression plasmid were constructed,and transfected into glioma cell lines:U87,U251 and LN229.Effects of hsacirc0008278 were observed on glioma cell proliferation,clonal formation and invasion and migration.Result:1.The sequencing results showed that there were 31 circRNAs differentially expressed between normal brain tissue and glioma tissues,13 of which were elevated in glioma tissues and 18 were decreased.GO analysis and KEGG pathway analysis revealed that differentially expressed circRNAs involved in multiple biological pathways and signaling pathways.2.Expression of 5 randomly selected differentially expressed circRNAs in 6 normal brain tissues and 45 glioma tissues was verified via RT-qPCR.Results showed that expression level of hsacirc0008278 was significantly decreased in glioma,and expression level of hsacirc0008278 was positively correlated with progression-free survival?PFS?and overall survival?OS?of glioma patients·3.After overexpression of hsacirc0008278 in glioma cell line,the proliferation,clonal formation and invasion and migration ability of glioma were decreased significantly,while knockdown expression of hsacirc0008278 had the opposite effect on the biological characteristics of glioma.Conclusion:There are differentially expressed circRNAs in glioma.These circRNAs are involved in various biological processes such as proliferation,differentiation and signal transduction in cells,which play an important role in regulating the development of glioma.The expression level of hsacirc0008278 was significantly decreased in glioma,and the low expression of hsacirc0008278 was significantly associated with poor prognosis of glioma patients.In vitro,to interfere with the expression of hsacirc0008278 can affect the biological characteristics of glioma,confirming that hsacirc0008278 plays an important role in the development of glioma,which provides a new target for gene therapy of glioma. |
PDF Full Text Request