临床儿科杂志 ›› 2015, Vol. 33 ›› Issue (11): 974-.doi: 10.3969 j.issn.1000-3606.2015.11.014

• 实验研究 • 上一篇    下一篇

miR-125a-5P 在髓母细胞瘤中的异常表达及其抑癌作用

楚冬梅,刘晓智,严艳萍,高心静,刘翠苹,李艳霞,姜忠敏,姚玲   

  1. 天津市第五中心医院(天津 300450)
  • 收稿日期:2015-11-15 出版日期:2015-11-15 发布日期:2015-11-15
  • 通讯作者: 姚玲 E-mail:yao-ling1960 @hotmail.com
  • 基金资助:
    天津市滨海新区医药卫生科研基金项目(No.2012BWKZ003)

Abnormal expression of miR-125a-5P in medulloblastoma and its role as tumor suppressor

CHU Dongmei, LIU Xiaozhi, YAN Yanping, GAO Xinjing, LIU Cuiping, LI Yanxia, JIANG Zhongmin, YAO Ling   

  1. The Fifth Central Hospital of Tianjin, Tianjin 300450, China
  • Received:2015-11-15 Online:2015-11-15 Published:2015-11-15

摘要: 目的 探讨miR-125a-5P在髓母细胞瘤中的表达特点并分析其抑癌作用。方法 采用逆转录PCR方法检测58份髓母细胞瘤组织及2个髓母细胞瘤株D341细胞和Daoy细胞中miR-125a-5p表达水平;瞬时转染D341细胞miR-125a-5P寡核苷酸,免疫荧光方法检测Ki-67核增殖活性,Transwell实验检测细胞迁移力,克隆形成实验检测瘤细胞致瘤潜能;荷瘤鼠模型检测miR-125a-5p转染对体内瘤细胞的抑制效果。结果 miR-125a-5P在儿童髓母细胞瘤组织中的相对表达量明显高于正常儿童脑组织,差异有统计学意义(t=17.55,P<0.01);miR-125a-5P在神经干细胞、髓母细胞瘤株D341细胞、Daoy细胞、神经胶质细胞和神经元细胞的相对表达量差异有统计学意义(F=9.44,P=0.002);miR-125a-5P的表达量在神经胶质细胞和神经元细胞中较高,分别高于神经干细胞、髓母细胞瘤株D341细胞和Daoy细胞中的相对表达量,差异有统计学意义(P均<0.05)。与对照组和无义组比较,miR-125a-5p组瘤细胞增殖速度减慢,瘤细胞迁移力降低,肿瘤克隆能力减弱,荷瘤鼠肿瘤体积小,差异均有统计学意义(P<0.05)。结论 miR-125a-5P在髓母细胞瘤中表达降低或缺失,可能是潜在的抑癌基因之一。

Abstract:  Objectives To study the expression characteristics of miR-125a-5P in medulloblastoma, and its role as tumor suppressor. Methods The miR-125a-5p level was detected by stem loop Real-time RT-PCR in 58 cases of medulloblastoma tissues and 2 medulloblastoma cell lines, D341 and Daoy cells. miR-125a-5P oligonucleotide was transfected to D341 cells, then Ki-67 expression was detected by immunofluorescence assay, cell migration was tested by transwell assay, and tumorigenicity was determined by clone formation assay. The inhibitory effect of miR-125a-5p on medulloblastoma cells in vivo was detected in tumor bearing mice model. Results The relative expression level of miR-125a-5P in medulloblastoma tissues was significantly higher than that of normal children's brain tissue (t=17.545, P<0.01), while the expression level of miR-125a-5P in 2 medulloblastoma cells were significantly lower than other control cells (F=9.437, P<0.01). The expression level of miR-125a-5P was significantly higher than that of the neural stem cells, D341 and Daoy cells, and the difference was statistically significant (P<0.05). Compared with the control group and the non-sense group, the ability of proliferation, migration and clone of cells in miR-125a-5p group obviously declined (P <0.05), as well as the tumor volume in mice (P<0.05). Conclusions The expression of miR-125a-5P in medulloblastoma was reduced or absent, and was supposedly a tumor suppressor gene.