Function And Mechanism Of Long Noncoding RNA BCYRN1 In Glioma

Objective To investigate the expression of long non-coding(Lnc RNA)BCYRN1 in glioma tissues and glioma cell lines,as well as the effect on the proliferation,apoptosis,invasion and migration ability of glioma cells.Further study of its possible biological mechanism will provide a theoretical basis for the clinical diagnosis,treatment and prognosis of gliomas.Methods High-throughput sequencing of 3 glioma tissue samples with 3 normal brain tissue samples as reference;analysis of differentially expressed Lnc RNAs in sequencing results using bioinformatics methods;111 cases of glioma tissues surgically resected from March 2018 to August 2019 were collected,and the expression level of Lnc RNA BCYRN1 was detected by QPCR;adhesive culture of glioma cell lines,QPCR was used to detect the expression level of Lnc RNA BCYRN1 in glioma cell lines;transfection of glioma cell lines T98 G and U251 with si RNA or plasmid to change the expression of Lnc RNA BCYRN1;CCK-8 cell proliferation assay to detect glioma cell proliferation;flow cytometry to detect glioma cell apoptosis;Transwell invasion assay to detect glioma cell invasion;Cell Wound Healing assay to detect glioma cell migration ability;Westernblotting method to detect protein expression changes in glioma cells;coimmunoprecipitation experiments and dual luciferase reports verified the binding site of Lnc RNA BCYRN1.Results Compared with the control group,the expression level of Lnc RNA BCYRN1 in glioma tissue was reduced(P<0.0001);compared with HEB cells,the glioma cell lines T98 G,U87,U251,and U373 Lnc RNA BCYRN1 expression levels were reduced(P<0.05);48 hours after transfection of si RNA with liposome(Lipofectamine 3000),realtime fluorescent quantitative polymerase chain reaction(QPCR)detection showed that the expression levels of Lnc RNA BCYRN1 in T98 G cells and U251 cells in si-1 and si-2 groups were lower than those in the control group.After plasmid transfection,the expression level of Lnc RNA BCYRN1 was higher than that of the control group(P<0.05);24,48,and 72 hours after transfection,microplate reader analysis showed that the proliferation ability of T98 G cells and U251 cells in the si-1 and si-2 groups was higher than that in the control group,and the cell proliferation activity was weakened after transfection of the plasmid(P<0.05);48 hours after transfection,flow cytometry showed that the apoptosis rates of T98 G cells and U251 cells in si-1 and si-2 groups were lower than those in the control group,and that in the plasmid group increased(P<0.05);48h after transfection,the number of T98 G and U251 cells passing through the chamber in the si-1 and si-2 groups was more than that in the control group,while the number of cell invasion in the plasmid group decreased(P<0.05);twelve and twenty-four hours after transfection,T98 G and U251 cells in the si-1 and si-2 groups were healed more than the control group,while the plasmid group migrated more slowly(P<0.05);AGO2 coimmunoprecipitation experiment and dual luciferase report confirmed that Lnc RNA BCYRN1 competitively binds to mi R-619-5p(P<0.05);Western blot results showed that mi R-619-5p reversely regulates the expression of CUEDC2.Conclusions(1)Lnc RNA BCYRN1 is under-expressed in glioma tissues and cell lines,and overexpression of Lnc RNA BCYRN1 can significantly inhibit glioma cell proliferation,invasion and migration,and promote cell apoptosis,indicating that Lnc RNA BCYRN1 is involved in the development of glioma play an important role.(2)Lnc RNA BCYRN1 competes with mi R-619-5p to regulate the expression of the tumor suppressor protein CUEDC2.(3)Lnc RNA BCYRN1 / mi R-619-5p / CUEDC2 axis inhibits the occurrence and development of gliomas,providing potential targets for the early diagnosis and treatment of gliomas.