lncRNA ZNF674-AS1过表达对胶质瘤U87细胞增殖、侵袭和迁移的影响

Effects of over-expression of lncRNA ZNF674-AS1 on proliferation,invasion and migration of U87 glioma cells

目的探讨长链非编码RNA(lncRNA)ZNF674-AS1过表达对胶质细胞瘤增殖、侵袭、迁移的影响。方法体外培养正常星形胶质细胞(HA1800)和胶质瘤细胞(A172、U251、U87、U373),RT-PCR检测lncRNA ZNF674-AS1表达水平。将ZNF674-AS1 mimics转染U87细胞上调ZNF674-AS1表达,以转染阴性对照序列为对照,CCK8法检测细胞增殖能力,Transwell实验检测细胞侵袭和迁移能力。Starbase软件预测lncRNA ZNF674-AS1靶基因并应用双荧光素酶报告基因实验验证。结果与正常星形胶质细胞(HA1800)比较,胶质瘤细胞(A172、U251、U87、U373)lncRNA ZNF674-AS1的表达水平均明显降低(P<0.05),其中U87细胞表达水平最低。上调U87细胞lncRNA ZNF674-AS1表达,明显抑制U87细胞增殖、侵袭、迁移能力(P<0.05)。Starbase软件预测显示lncRNA ZNF674-AS1与性别决定区Y框蛋白9(SOX9)基因有结合位点,双荧光素酶报告基因实验结果显示,SOX9基因是lncRNA ZNF674-AS1靶基因。上调U87细胞lncRNA ZNF674-AS1表达的同时沉默SOX9基因表达,明显增强U87细胞增殖、侵袭和迁移能力(P<0.05)。结论胶质瘤lncRNA ZNF674-AS1呈低表达,可能通过靶向下调SOX9基因表达,促进胶质瘤细胞增殖、侵袭和迁移。

Objective To investigate the effect of over-expression of lncRNA ZNF674-AS1 on proliferation,invasion and migration of U87 glioma cells.Methods The expression levels of lncRNA ZNF674-AS1 were detected by RT-PCR in normal astrocytes(HA1800)and glioma cells(A172,U251,U87,U373).ZNF674-AS1 mimics were transfected into U87 cells to up-regulate expression of lncRNA ZNF674-AS1,with negative sequence as control.Cell proliferation was detected by CCK8 assay,and cell invasion and migration were detected by Transwell assay.lncRNA ZNF674-AS1 target genes were predicted by Starbase software and verified by dual luciferase reporter gene assay.Results Compared with normal astrocytes(HA1800),the expression levels of lncRNA ZNF674-AS1 in glioma cells(A172,U251,U87,and U373)were significantly decreased(P<0.05),and the expression level of lncRNA ZNF674-AS1 in U87 cells was the lowest.Over-expression of lncRNA ZNF674-AS1 significantly inhibited the proliferation,invasion and migration of U87 cells(P<0.05).Starbase software predicted that lncRNA ZNF674-AS1 had a binding site with the sex determining region Y-box protein 9(SOX9)gene,and the double luciferase reporter gene experiment showed that SOX9 gene was the target gene of lncRNA ZNF674-AS1.Over-expression of lncRNA ZNF674-AS1 combined with silence expression of SOX9 significantly enhanced the proliferation,invasion and migration of U87 cells(P<0.05).Conclusions The expression of lncRNA ZNF674-AS1 is down-regulated in glioma,which may promote the proliferation,invasion and migration of glioma cells by down-regulating expression of SOX9 gene.