The Research Of The Functional Interaction Between LncRNAs And NSPc1 In U87 Glioma Cells

Objective: Recent studies have found that Polycomb Group(PcG)proteins play an epigenetic repressive role in transcription and are closely related to the occurrence and development of glioma.Meanwhile,the abnormal expression of long noncoding RNAs(lncRNAs)in glioma is associated with grade and classification of tumor.The expression of lncRNAs in differentiated U87 glioma cells and the expression change of lncRNAs with the up-regulated or down-regulated expression of NSPc1 in U87 glioma cells were analyzed in order to identify the possible functional interaction between lncRNAs and NSPc1 in U87 glioma cells.Methods: First of all,the expressions of NSPc1 and the lncRNAs selected which may interact with NSPc1 were detected by Real-time quantitative PCR in malignant glioma cells such as H4,U251,U87 cells;Then U87 glioma cells were induced to differentiation with 10-6mol/L concentration of retinoic acid.The expressions of NSPc1 and candidate lncRNAs were detected in several differentiation stage.Moreover,the expressions of candidate lncRNAs were tested after raising or knocking down NSPc1 expression comparied with the expressions of candidates in normal U87 cell.Then,the functional interactions between lncRNAs and NSPc1 were identified by applying MTT assay and RNA binding protein Immunoprecipitation(RIP).Result:1 The expression of NSPc1 was positively related to the malignant degree in different glioma cell lines.The marked inequality expression among different lncRNAs exsited in glioma cell lines.2 In the retinoic acid induced differentiation process(0~10days)of U87 glioma cells,the expression level of NSPc1 decreased initially and achieves the lowest point at the sixth day;but rised in the following days till the tenth day;The expression of lncRNAs decreased in the differentiation stage.3 The expression levels of lncRNAs changed after over-expressing or down-regulating the expression of NSPc1.What is more,the expression of MALAT1 and ANRIL significantly changed.4 Knocking down the expression of MALAT1 and ANRIL inhibited the growth of U87 glioma cells and raised the ratio of cell apoptosis.5 The interaction between MALAT1,ANRIL and NSPc1 are identified by RIP essays and the enrichment of NSPc1-ANRIL was positively correlated with the expression of NSPc1 in the process of U87 glioma cell differentiation.Conclusion:1 NSPc1 expresses in glioma and is closely relative to the malignant degree of gliomas.2 MALAT1,ANRIL can promote the growh of U87 glioma cells and inhibit apoptosis of U87 glioma cells.3 NSPc1 interactes with MALAT1 and ANRIL,and the ANRIL-NSPc1 functional interaction may play an important role in the process of U87 glioma cell differentiation.