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

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。

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

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.