SNHG15-lncRNA ceRNA网络靶向miR-451a促进胶质瘤增殖

SNHG15-lncRNA CeRNA Network Targets MiR-451a to Promote the Proliferation of Glioma

目的:探索SNHG15-lncRNA ceRNA网络靶向miR-451a促进胶质瘤增殖的作用机制。方法:qRT-PCR检测SNHG15和miR-451a在临床组织标本、人脑神经胶质细胞系HEB和神经胶质瘤细胞系(U87、U251和Hs683)的表达量中的表达量;双荧光素酶报告基因检验、qRT-PCR和Pull down检测SNHG15-miR-451a轴靶向调控关系。分别将SNHG15小干扰RNA(siRNA)、阴性对照序列(NC)、miR-451a抑制剂(miR-451a inhibitor)对U87细胞进行转染。细胞随机分为转染阴性对照序列(siNC)组、转染SNHG15 siRNA(siSNHG15)组、转染SNHG15 siRNA和miR-451a inhibitor(siSNHG15+miR-451a inhibtor)组。Tranwell细胞侵袭、细胞划痕和Tunel细胞凋亡分别检测SNHG15和miR-451a对U87细胞侵袭、迁移及凋亡的调控;构建裸鼠皮下移植瘤模型检测肿瘤生长及体内细胞增殖情况。结果:与癌旁组织及HEB细胞相比,胶质瘤组织及胶质瘤细胞系中的SNHG15表达显著增高,而miR-451a表达显著降低(均P<0.05)。双荧光素酶报告基因结果显示转染miR-451a inhibitor后SNHG15荧光素酶活性显著增加(P<0.05);Pull down生物素化实验结果显示miR-451a降低U87细胞中SNHG15的表达水平(P<0.05);而转染SNHG15质粒后miR-451a水平表达降低(P<0.05)。SNHG15显著抑制,而miR-451a inhibitor显著促进U87细胞侵袭和迁移距离及细胞凋亡率(均P<0.05)。SNHG15组移植瘤重量显著高于较NC组,而miR-451a组移植瘤重量显著低于miR-NC组(均P<0.05)。siSNHG15组PCNA表达低于siNC组及siSNHG15+miR451a inhibitor组(均P<0.05);miR-451a inhibitor组PCNA表达高于NC inhibitor组及siSNHG15+miR451a inhibitor组(均P<0.05)。结论:SNHG15竞争性结合miR-451a促进胶质瘤细胞侵袭和迁移,下调SNHG15-miR-451a轴可能作为治疗胶质瘤的新靶点。

Objective:To explore the mechanism of SNHG15-lncRNA ceRNA network to promote glioma proliferation by targeting miR-451a.Methods:The expression levels of SNHG15 and miR-451a in clinical tissue samples,human glial cell lines HEB,and glioma cell lines(U87,U251 and Hs683)were detected by qRT-PCR.Dual luciferase reporter gene assay,qRT-PCR and Pull down were used to detect the targeted regulation of SNHG15-miR-451a axis.U87 cells were transfected with SNHG15 small interfering RNA(siRNA),negative control sequence(NC)and miR-451a inhibitor.The cells were randomly divided into siNC group,siSNHG15 group and siSNHG15+miR-451a inhibitor group.The regulation of invasion,migration and apoptosis of U87 cells by SNHG15 and miR-451a were detected by Transwellcell invasion,cell scratching and Tunel cell apoptosis,respectively.A subcutaneous transplantation tumor model of glioma in nude mice was constructed to detect tumor growth and cell proliferation in vivo.Results:Compared with the adjacent tissues and HEB cells,the expression of SNHG15 in glioma tissues and glioma cell lines was significantly increased,while the expression of miR-451a was significantly decreased(all P<0.05).Dual luciferase reporter gene results showed that SNHG15 luciferase activity increased significantly after transfection with miR-451a inhibitor(P<0.05).Pull down biotinylation assay showed that miR-451a decreased the expression level of SNHG15 in U87 cells(p<0.05),while the expression of miR-451a was decreased after transfection with SNHG15 plasmid(P<0.05).SNHG15 significantly inhibited,while miR-451a inhibitor significantly promoted the invasion,migration distance and apoptosis rate of U87 cells(all P<0.05).The SNHG15 group had a significantly higher graft weight than the NC group,while the miR-451a group had a significantly lower graft weight than the miR-NC group(both P<0.05)The expression of PCNA in siSNHG15 group was lower than in siNC group and siSNHG15+miR-451a inhibitor group(both P<0.05).The expression of PCNA in miR-451a inhibitor group was higher than in NC inhibitor group and siSNHG15+miR-451a inhibitor group(all P<0.05).Conclusions:SNHG15 competitively binds miR-451a to promotes the invasion and migration of glioma cell,and inhibits apoptosis.Down-regulating the SNHG15-miR-451a axis might serve as a new therapeutic target for glioma treatment.