Wang Yunling, Lin Yihai
Departments of Neurology.
Neurosurgery, Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang Province, People's Republic of China.
Neuroreport. 2018 Sep 26;29(14):1166-1173. doi: 10.1097/WNR.0000000000001089.
This work aimed to investigate the inter-regulatory functions of hsa-mir-127 and replication initiator 1 (REPIN1) on the proliferation and metastasis of glioma cells. The in-silico data on the implication of hsa-mir-127 and REPIN1 in glioma were retrieved from The Cancer Genome Atlas (TCGA). The expression levels of hsa-mir-127 and REPIN1 mRNA were determined by qRT-PCR, whereas Western blot was used to detect REPIN1 protein expression in glioma cell lines. The proliferation of glioma cells was determined by means of the MTT assay, whereas the transwell assay was employed for assessing the extent of cell migration and invasion. The interaction among REPIN1 and hsa-mir-127 was checked using the luciferase reporter assay. The expression of hsa-mir-127 was markedly increased in clinical data obtained from TCGA and in glioma cells compared with normal tissues and control cells, respectively. Increased expression of hsa-mir-127 and decreased expression of REPIN1 were both associated with poor overall survival. Moreover, hsa-mir-127 overexpression noticeably promoted proliferation, inhibited apoptosis and increased the invasive and migratory capacities of glioma cells. Inverse effects were found with hsa-mir-127 antisense inhibitor. Interestingly, overexpression of hsa-mir-127 downregulated REPIN1 expression, and luciferase reporter assay showed that the tumorigenesis effect of hsa-mir-127 requires, in part, its direct targeting of REPIN1. In conclusion, the hsa-mir-127/REPIN1 pathway is involved in gliomas and could be a potential therapeutic target.
这项研究旨在探讨人源微小RNA-127(hsa-mir-127)与复制起始因子1(REPIN1)对胶质瘤细胞增殖和转移的相互调控作用。从癌症基因组图谱(TCGA)中检索了hsa-mir-127和REPIN1在胶质瘤中的计算机分析数据。通过qRT-PCR测定hsa-mir-127和REPIN1 mRNA的表达水平,而采用蛋白质印迹法检测胶质瘤细胞系中REPIN1蛋白的表达。通过MTT法测定胶质瘤细胞的增殖,而采用Transwell实验评估细胞迁移和侵袭程度。使用荧光素酶报告基因检测法检测REPIN1与hsa-mir-127之间的相互作用。与正常组织和对照细胞相比,分别从TCGA获得的临床数据和胶质瘤细胞中,hsa-mir-127的表达明显增加。hsa-mir-127表达增加和REPIN1表达降低均与总体生存率差有关。此外,hsa-mir-127过表达显著促进增殖、抑制凋亡并增加胶质瘤细胞的侵袭和迁移能力。hsa-mir-127反义抑制剂则产生相反的效果。有趣的是,hsa-mir-127过表达下调REPIN1表达,荧光素酶报告基因检测表明,hsa-mir-127的致瘤作用部分需要其直接靶向REPIN1。总之,hsa-mir-127/REPIN1通路参与胶质瘤的发生,可能是一个潜在的治疗靶点。