Sun Guan, Shi Lei, Yan Shushan, Wan Zhengqiang, Jiang Nan, Fu Linshan, Li Min, Guo Jun
Department of Neurosurgery, Fourth Affiliated Hospital of Nantong University, First Hospital of Yancheng, Yancheng 224001, China.
Department of Neurosurgery, The First People's Hospital of Kunshan Affiliated with Jiangsu University, Suzhou 215300, China.
Biomed Res Int. 2014;2014:687826. doi: 10.1155/2014/687826. Epub 2014 Jun 4.
To investigate the role and mechanism of miR-15b in the proliferation and apoptosis of glioma.
The miR-15b mimics were transfected into human glioma cells to upregulate the miR-15b expression. Cyclin D1 was determined by both western blotting analysis and luciferase reporter assay. Methylthiazol tetrazolium (MTT) and flow cytometry were employed to detect the cell proliferation, cell cycle, and apoptosis.
Overexpression of miR-15b inhibits proliferation by arrested cell cycle progression and induces apoptosis, possibly by directly targeting Cyclin D1. Both luciferase assay and bioinformatics search revealed a putative target site of miR-15b binding to the 3'-UTR of Cyclin D1. Moreover, expression of miR-15b in glioma tissues was found to be inversely correlated with Cyclin D1 expression. Enforced Cyclin D1 could abrogate the miR-15b-mediated cell cycle arrest and apoptosis.
Our findings identified that miR-15b may function as a glioma suppressor by targeting the Cyclin D1, which may provide a novel therapeutic strategy for treatment of glioma.
探讨miR-15b在胶质瘤增殖和凋亡中的作用及机制。
将miR-15b模拟物转染到人胶质瘤细胞中以上调miR-15b表达。通过蛋白质印迹分析和荧光素酶报告基因检测来测定细胞周期蛋白D1。采用甲基噻唑基四氮唑（MTT）法和流式细胞术检测细胞增殖、细胞周期和凋亡情况。
miR-15b的过表达通过阻滞细胞周期进程抑制增殖并诱导凋亡，可能是通过直接靶向细胞周期蛋白D1。荧光素酶检测和生物信息学搜索均揭示了miR-15b与细胞周期蛋白D1的3'-非翻译区结合的一个假定靶位点。此外，发现胶质瘤组织中miR-15b的表达与细胞周期蛋白D1的表达呈负相关。过表达细胞周期蛋白D1可消除miR-15b介导的细胞周期阻滞和凋亡。
我们的研究结果表明，miR-15b可能通过靶向细胞周期蛋白D1发挥胶质瘤抑制因子的作用，这可能为胶质瘤的治疗提供一种新的治疗策略。
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