敲减lncRNA RP1-261G23.7抑制胶质瘤细胞增殖和迁移及其机制探讨

Knockdown of lncRNA RP1-261G23.7 inhibits the proliferation and migration of glioma cells and its mechanism

目的:探讨长链非编码RNA(long non-coding RNA,lncRNA)RP1-261G23.7对人胶质瘤细胞增殖及迁移的影响及可能的调控机制。方法:采用GEPIA数据库分析RP1-261G23.7在胶质瘤组织中的相对表达。采用qRT-PCR检测RP1-261G23.7在4种胶质瘤细胞系(U87MG、SNB-19、U251、LN382)中的相对表达。采用Lipofectamine3000向胶质瘤细胞U87MG中单独转染si-RP1-261G23.7质粒(si-RP1-261G23.7组)和si-NC对照质粒(si-NC组)。采用CCK-8实验和细胞划痕实验检测转染后U87MG细胞的增殖及迁移能力。采用生物信息学、qRT-PCR和双荧光素酶报告基因实验研究RP1-261G23.7和miR-525-5p表达的关系。Western blotting检测NF-κB信号通路蛋白的表达。结果:与正常组织相比,RP1-261G23.7在胶质瘤组织中表达上调(P<0.01)。与人脑星型胶质正常细胞系(HEB)相比,RP1-261G23.7在四种胶质瘤细胞系中表达均上调(P<0.01),U87MG细胞中RP1-261G23.7相对表达量最高(P<0.01)。与si-NC组相比,敲减RP1-261G23.7显著抑制了U87MG细胞的增殖(P<0.05)和划痕愈合(P<0.01)。RP1-261G23.7能够直接互补结合miR-525-5p(P<0.01)。与si-NC组相比,敲减RP1-261G23.7表达显著促进了U87MG细胞中miR-525-5p的表达(P<0.01),NF-κB信号通路蛋白表达显著下降(P<0.01)。结论:胶质瘤组织和细胞系中RP1-261G23.7表达明显上调,敲减RP1-261G23.7通过促进miR-525-5p表达、干扰NF-κB信号通路活化,抑制胶质瘤U87MG细胞的增殖和迁移,可能为胶质瘤的靶向治疗开辟新的路径。

Objective:To explore the effect of long non-coding RNA(lncRNA)RP1-261 G23.7 on the proliferation and migration of human glioma cells and the possible regulatory mechanism.Methods:The GEPIA database was used to analyze the relative expression of RP1-261 G23.7 in glioma tissues.qRT-PCR was used to detect the relative expression of RP1-261 G23.7 in 4 glioma cell lines(U87 MG,SNB-19,U251,LN382).Lipofectamine3000 was used to separately transfect the si-RP1-261 G23.7 plasmid(si-RP1-261 G23.7 group)and the si-NC control plasmid(si-NC group)into the glioma cell U87 MG.CCK-8 experiment and cell scratch experiment were used to detect the proliferation and migration ability of U87 MG cells after transfection.Bioinformatics,qRT-PCR and dual luciferase reporter gene experiments were used to study the relationship between RP1-261 G23.7 and miR-525-5 p expression.Western blotting was used to detect the expression of NF-κB signaling pathway proteins.Result:Compared with normal tissues,RP1-261 G23.7 was up-regulated in glioma tissues(P<0.01).Compared with human brain astrocyte normal cell line(HEB),the expression of RP1-261 G23.7 was up-regulated in glioma cell lines(P<0.01),and the relative expression of RP1-261 G23.7 was the highest in U87 MG cells(P<0.01).Compared with the si-NC group,knockdown of RP1-261 G23.7 significantly inhibited the proliferation of U87 MG cells(P<0.05)and scratch healing(P<0.01).RP1-261 G23.7 could directly complement miR-525-5 p(P<0.01).Compared with the si-NC group,knockdown of RP1-261 G23.7 significantly promoted the expression of miR-525-5 p in U87 MG cells(P<0.01),and the protein expression of NF-κB signaling pathway decreased significantly(P<0.01).Conclusion:The expression of RP1-261 G23.7 was significantly up-regulated in glioma tissues and cell lines.Knockdown of RP1-261 G23.7 promotes the expression of miR-525-5 p,interferes with the activation of the NF-κB signaling pathway,and inhibits the proliferation and migration of glioma U87 MG cells.RP1-261 G23.7 may open up a new path for targeted therapy of glioma.