| Objection: To explore the function of lnc RNA RP11-46C24.7 in glioma and glioma stem cells.Methods: 1.The glioma stem cells were enriched by serum-free medium within growth factors EGF,b FGF and B27.2.By reviewing the relevant literature,q RT-PCR was used to screen lnc RNAs with differential expression in glioma and glioma stem cells,and lnc RNA RP11-46C24.7 with obvious expression difference was selected.The role of lnc RNA RP11-46C24.7 in glioma and glioma stem cells was investigated.3.Lnc RNA RP11-46C24.7 low stable expression was constructed in U87 and U251 cells by lentiviral infection.4.The effects of lnc RNA RP11-46C24.7 on the proliferation and migration of glioma cells were detected by CCK8,clonality formation,cell cycle,apoptosis,transwell and cell scratches.5.The effects of lnc RNA RP11-46C24.7 on the stemness,sphere formation ability and growth of glioma stem cells were detected by q RT-PCR and sphere formation experiments.Results: 1.The glioma stem cells were successfully enriched by serum-free medium.2.The expression of lnc RNA RP11-46C24.7 in GSCs was higher than that in glioma cells by q RT-PCR.3.Lnc RNA RP11-46C24.7 low stable expression was successfully constructed in U87 and U251 cells by lentiviral infection.4.Stable RNA interference-mediated knockdown of lnc RNA RP11-46C24.7 in U87 and U251 cells significantly reduced proliferation ability of cells by CCK8 and colony formation assay.5.Knockdown of lnc RNA RP11-46C24.7,the percentage of glioma cells in G0/G1 phase was significantly increased,and the percentage of cells in S phase was decreased by flow cytometry analysis.6.Flow cytometry was used to detect the down-regulation of lnc RNA RP11-46C24.7 expression,which increased the number of glioma cell apoptosis about 3 times.7.Down-regulated the expression of lnc RNA RP11-46C24.7,reducing the migration capacity of glioma cells by cell scratches and transwell assays.8.Low-expression lnc RNA RP11-46C24.7 U87 and U251 cell was successfully enriched low expression of GSCs in serum-free.9.Low expression of lnc RNA RP11-46C24.7,which reduced the expression of CD133 and Nestin on the biomarker of stem cells by q PCR.10.Low expression of lnc RNA RP11-46C24.7 reduced stem cell diameter by sphere formation experiments.Conclusion: The results showed that stable RNA interference-mediated knockdown of lnc RNA RP11-46C24.7 in U87 and U251 cells obviously decreased proliferation ability of cells,accelerated the apoptosis of cells,and suppressed the migration of cells.Down-regulation of lnc RNA RP11-46C24.7 inhibited stemness and slowed the growth and self-renewal of glioma stem cells by sphere formation experiments. |
PDF Full Text Request