Wang Jin, Liu Xiaoyang, Yan Changsheng, Liu Jie, Wang Songtao, Hong Yongzhi, Gu Aihua, Zhao Peng
Department of Neurosurgery.
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing.
Onco Targets Ther. 2017 Aug 28;10:4251-4260. doi: 10.2147/OTT.S130365. eCollection 2017.
The long-noncoding RNAs (lncRNAs) are identified as new crucial regulators of diverse cellular processes in glioblastoma (GBM) tissues. However, the expression pattern and biological function of lncRNAs remain largely unknown. Here, for the first time, the effects of lncRNA lymphoid enhancer-binding factor 1 antisense RNA 1 () on GBM progression both in vitro and in vivo are investigated.
Expression profiles of in GBM specimens were investigated by bioinformatics analyses. expression in GBM tissues was detected using a quantitative polymerase chain reaction. expression was inhibited by transfecting the -specific small interfering RNAs (siRNAs) and stable cell lines established were inhibited by transfecting si- viruses. The Cell Counting Kit-8, ethynyl deoxyuridine, and colony formation assay were used to examine proliferation function. The flow cytometry detected cell-cycle change and apoptosis. Migration effects were detected by a Transwell assay. The tumor xenografts and immunohistochemistry were performed to evaluate tumor growth in vivo.
In this study, expression was found significantly upregulated in GBM specimens compared with normal tissues. The 5-year overall survival in GBM patients from The Cancer Genome Atlas with high expression of was inferior to that with low expression. It was confirmed that expression of was higher in GBM tissues than normal ones. Knockdown of significantly inhibited the malignancy of GBM cells, including proliferation and invasion, and promoted cell apoptosis. The result of Western blot assays indicated that knockdown of -mediated tumor suppression in GBM cells may be via the reduction of ERK and Akt/mTOR signaling activities. Finally, the in vivo experiment also demonstrated that knockdown inhibited the growth-promoting effect of of U87 cells.
Our result indicated that lncRNA acts as an oncogene in GBM and may be a pivotal target for this disease.
长链非编码RNA（lncRNAs）被确定为胶质母细胞瘤（GBM）组织中多种细胞过程的新关键调节因子。然而，lncRNAs的表达模式和生物学功能仍 largely未知。在此，首次研究了lncRNA淋巴样增强因子1反义RNA1（）对GBM体外和体内进展的影响。
通过生物信息学分析研究GBM标本中的表达谱。使用定量聚合酶链反应检测GBM组织中的表达。通过转染特异性小干扰RNA（siRNAs）抑制表达，并通过转染si病毒建立稳定细胞系。使用细胞计数试剂盒-8、乙炔基脱氧尿苷和集落形成试验检测增殖功能。流式细胞术检测细胞周期变化和凋亡。通过Transwell试验检测迁移作用。进行肿瘤异种移植和免疫组织化学以评估体内肿瘤生长。
在本研究中，发现GBM标本中的表达与正常组织相比显著上调。来自癌症基因组图谱的高表达GBM患者的5年总生存率低于低表达患者。证实GBM组织中的表达高于正常组织。敲低显著抑制GBM细胞的恶性程度，包括增殖和侵袭，并促进细胞凋亡。蛋白质印迹分析结果表明，敲低介导的GBM细胞肿瘤抑制可能是通过降低ERK和Akt/mTOR信号活性。最后，体内实验也表明敲低抑制了U87细胞的生长促进作用。
我们的结果表明lncRNA在GBM中起癌基因作用，可能是这种疾病的关键靶点。
