Function Of LncRNA RP4-758J24.5-S In Glioma And Glioma Stem Cells

Objection To explore the function and mechanism of LncRNA RP4-758J24.5-S in glioma cells and glioma stem cells(GSCs).Methods 1.Enriching U87 and U251 tumor stem cells by serum-free starvation culture.2.By searching the literature,some LncRNAs with different expression levels in glioma cancer tissues and adjacent tissues were obtained.The expression differences of these LncRNAs in U87 and U87 stem cells,U251 and U251 stem cells were verified by PCR and QPCR.LncRNAs with significant differences in expression were obtained,and LncRNA RP4-758J24.5-S used in the study was screened out.3.The SiRNA used to silence LncRNA RP4-758J24.5-S was designed in vitro,and the effect of down-regulation of LncRNA RP4-758J24.5-S on the proliferation of U87 and U251 in vitro was detected by transient transfection.4.Construction of a lentiviral stable cell line with low expression of LncRNA RP4-758J24.5-S.5.Using CCK8,plate clone formation assay,cell scratch assay,transwell,Annexin VFITC/PI double staining,PI staining to detect LncRNA RP4-758J24.5-S low expression of lentivirus stable cell line proliferation ability,colony forming ability,migration abi lity Changes in cell cycle and apoptosis.6.Using QPCR and stem cell globule assay to detect the dryness,self-renewal ability and globular ability of U87 and U251 stem cell stably expressing LncRNA RP4-758J24.5-S.Results 1.This study successfully isolated glioma stem cells(GSCs,U87 GSCs and U251GSCs)from glioma cells(U87 and U251)by serum-free starvation-inducing culture method;2.PCR and QPCR results showed that the expression of LncRNA RP4-758J24.5-S in U87 s and U251 s was higher than that in U87 and U251 cells.3.Lentiviral stable cell line with low expression of LncRNA RP4-758J24.5-S was constructed.Down-regulation of LncRNA RP4-758J24.5-S can effectively reduce the cell proliferation ability,clone formation ability of U87 and U251,and significantly change the cell cycle and promote apoptosis.4.There was no significant difference in the down-regulation of LncRNA RP4-758J24.5-S on the migration ability of U87 and U251.5.After down-regulating the expression level of LncRNA RP4-758J24.5-S in GSCs cells,the diameter of tumor stem cells became smaller,the rate of tumor stem cell formation decreased,and the expression of cell dryness-related molecular markers was down-regulated.Conclusion Decreasing the expression of LncRNA RP4-758J24.5-S can significantly reduce the proliferation and clonality of U87 and U251,significantly affecting the cell cycle,causing G2/M phase arrest in U87 and U251 cells,and promoting apoptosis,but for The cell migration ability of U87 and U251 did not have much effect.Low expression of LncRNA RP4-758J24.5-S inhibits the proliferation of GSCs,stem cell globule formation and self-renewal,and reduces the dryness of GSCs.