摘要
目的观察非编码RNA00460靶向微小RNA(miRNA,miR)-539-性别决定基因相关HMG盒基因4(SOX4)对脑胶质瘤细胞增殖和迁移的影响.方法收集正常脑胶质和脑胶质瘤组织各20例,采用荧光定量聚合酶链反应(PCR)分析长链非编码RNA(lncRNA) 00460和miR-539表达水平和两者的关系.采用生物信息学分析lncRNA00460和miR-539之间的关系以及靶基因;在U251脑胶质瘤细胞株中构建lncRNA00460敲降细胞株和miR-539过表达细胞株,采用细胞计数试剂盒(CCK-8)和Transwell分析不同处理的脑胶质瘤细胞的增殖和迁移能力;采用异种移植瘤模型分析细胞在体内的增殖能力.结果与正常脑胶质细胞[(1.21±0.25)、(1.35±0.29)]比较,脑胶质瘤组织中lncRNA00460表达水平(3.41±0.51)显著升高,而miR-539表达水平(0.39±0.11)显著下调,差异均有统计学意义(t=3.189、4.208,P<0.05).生物信息学研究显示lncRNA00460调控着miR-539的表达水平,并进一步调控SOX4蛋白的表达.lncRNA00460敲降可显著抑制脑胶质瘤细胞的增殖和迁移,差异均有统计学意义(F=3.729、3.109,P<0.05).过表达miR-539显著增加了胶质瘤细胞的增殖和迁移,差异均有统计学意义(F=5.129、5.114,P<0.05).体内异种成瘤模型显示lncRNA00460敲降可显著抑制脑胶质瘤细胞在体内的生长,而过表达miR-539则显著促进肿瘤细胞的增殖,差异有统计学意义(F=5.194、3.104,P<0.05).结论 lncRNA00460通过靶向作用于miR-539/SOX4轴进而调节着脑胶质瘤细胞的增殖和迁移.
Objective To investigate the effects of non-coding RNA00460 targeting microRNA (miRNA, miR)-539-SRY-related high mobility group-box gene 4 (SOX4) on the proliferation and migration of glioma cells. Methods Twenty cases of normal brain glia and 20 cases of glioma were collected. The expression levels of lncRNA 00460 and miR-539 were analyzed by fluorescence quantitative polymerase chain reaction (PCR). Bioinformatics was used to analyze the relationship between lncRNA 00460 and miR-539 and target genes;knockdown cell line lncRNA 00460 and miR-539 overexpression cell line were constructed in U251 glioma cell line. cell counting kit-8 (CCK-8) and Transwell were used to analyze the proliferation and migration ability of glioma cells treated with different methods;xenograft tumor model was used to analyze the proliferation ability of glioma cells in vivo. Results Compared with normal brain glioma cells [(1.21±0.25),(1.35±0.29)], the expression of lncRNA 00460 (3.41±0.51) was significantly increased in glioma tissues, while the expression of microR-539 (0.39±0.11) was significantly decreased (t=3.189, 4.208, P<0.05). Bioinformatics studies have found that lncRNA 00460 regulates the expression of miR-539 and further regulates the expression of SOX4 protein. The knockdown of lncRNA 00460 significantly inhibited the proliferation and migration of glioma cells (F=3.729, 3.109, P<0.05). Overexpression of miR-539 significantly increased the proliferation and migration of glioma cells (F=5.129, 5.114, P<0.05). In vivo xenogeneic tumorigenesis model showed that knockdown of lncRNA 00460 significantly inhibited the growth of glioma cells in vivo, while over-expression of microRNA539 significantly promoted the proliferation of glioma cells (F=5.194, 3.104, P<0.05). Conclusionlnc RNA 00460 regulates the proliferation and migration of glioma cells by targeting the miR-539/SOX4 axis.
作者
常波
高飞
梁庆华
Chang Bo;Gao Fei;Liang Qinghua(Department of Neurosurgery,Anyang People’s Hospital,Anyang 455000,China;Department of Ophthalmology,Anyang People’s Hospital,Anyang 455000,China;Department of Neurosurgery,Henan People’s Hospital,Zhengzhou 450000,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第10期1802-1805,共4页
Chinese Journal of Experimental Surgery