国际肿瘤学杂志

国际肿瘤学杂志 ›› 2013, Vol. 40 ›› Issue (2): 149-152.

• 论著 • 上一篇    下一篇

反义miR-30a-5p抑制脑胶质瘤生长的体内实验研究

孙即奎, 贾志凡, 浦佩玉, 王广秀, 张安玲, 杨卫东   

  1. 300052 天津医科大学总医院神经病学研究所神经肿瘤室
  • 出版日期:2013-02-08 发布日期:2013-01-25
  • 通讯作者: 杨卫东，Email: yangweidongshine@sina.com E-mail:yangweidongshine@sina.com
  • 基金资助:

    国家自然科学基金（30872985）

Inhibitory effect of antisense miR-30a-5p on glioma cell growth in vivo

SUN  Ji-Kui, JIA  Zhi-Fan, PU  Pei-Yu, WANG  Guang-Xiu, ZHANG  An-Ling, YANG  Wei-Dong   

  1. Laboratory of Neuro-Oncology, Tianjin Neurological Institute, Tianjin Medical University General Hospital, Tianjin 300052, China
  • Online:2013-02-08 Published:2013-01-25
  • Contact: Corresponding author: YANG Wei-dong, E-mail: yangweidongshine@sina.com E-mail:yangweidongshine@sina.com

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被引次数

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摘要： 目的  探讨敲低微RNA(miR)-30a-5p表达对裸鼠荷人脑胶质瘤细胞U87移植瘤生长的抑制作用及其机制。方法 miR-30a-5p反义寡聚核苷酸(AS-miR-30a-5p)皮下注射治疗裸鼠皮下荷U87人脑胶质瘤模型。隔日测量肿瘤大小以评估其疗效。治疗结束后处死动物，剥离肿瘤组织，提取RNA和蛋白，采用Real-time PCR方法检测治疗后miR-30a-5p表达水平，采用石蜡切片苏木精-伊红染色、免疫组织化学染色及Western blot检测，评价治疗后肿瘤组织形态以及增殖、凋亡等生物学性状等相关指标的变化，包括隔蛋白7（SEPT7）、增殖细胞核抗原（PCNA）、细胞周期蛋白（cyclinD1）、基质金属蛋白酶-2（MMP-2）以及凋亡相关因子P53、bcl-2、caspase3等；应用TUNEL法检测肿瘤细胞凋亡。结果 AS-miR-30a-5p治疗组肿瘤生长速度及体积明显小于对照组与无义序列组，差异有统计学意义(F=7.167，P＜0.05)；其miR-30a-5p表达下调为对照组的0.297±0.0512；肿瘤细胞恶性生物学表型相关蛋白PCNA、cyclinD1等表达明显下调，P53、SEPT7、caspase3等肿瘤生长抑制因子表达上调；细胞凋亡数明显增加。结论 miR-30a-5p为靶点的治疗能有效抑制异种移植U87人脑胶质瘤生长，肿瘤恶性表型有明显逆转，可作为人脑胶质瘤靶向治疗的候选靶点。

关键词: 神经胶质瘤, 寡核苷酸类, 反义, 药物疗法, miR-30a-5p

Abstract: Objective To study the inhibitory effect of knocking down miR-30a-5p on the U87 human glioma xenograft growth and its possible mechanism. Methods Nude mice bearing subcutaneous U87 human glioblastoma were established and treated with miR-30a-5p antisense oligonucleotides (AS-miR-30a-5p) subcutaneous injection. Tumor size was measured every other day until the observation period ended. Researchers executed the animals after the treatment, stripped tumor tissues and extracted RNA and protein. Real-time PCR were conducted to detect the expression of miR-30a-5p. The histopathological characteristics and proliferation and apoptosis biological characters (including SEPT7, PCNA, cyclinD1, MMP-2, apoptosis related factor P53, bcl-2 and caspase3)were evaluated by HE and immunohistochemical staining, Western blot analysis respectively, and the cell apoptosis was detected by TUNEL method. Results In AS-miR-30a-5p treated group, the tumor growth was delayed and the final tumor volume was smaller than that in the control and scr-ODN treated group (F=7.167, P＜0.05), and the expression of miR-30a-5p was knocked down. The expression of PCNA、cyclinD1 were significantly down-regulated while P53、SEPT7 and caspase3 up-regulated. Apoptotic index was increased significantly. Conclusion As-miR-30a-5p suppresses the growth of U87 human gliomas xenografts significantly. Malignant phenotype of tumors are reversed to a considerable degree. Therefore, miR-30a-5p can be a candidate for targeted therapy of human glioma.

Key words: Giomas, Oligonucleotides, antisense, Drug therapy, miR-30a-5p