Nix和miR-520h对神经胶质瘤发生的影响及相互作用

doi:10.13418/j.issn.1001-165x.2014.05.019

中国临床解剖学杂志 ›› 2014, Vol. 32 ›› Issue (5): 590-594.doi: 10.13418/j.issn.1001-165x.2014.05.019

Nix和miR-520h对神经胶质瘤发生的影响及相互作用

胡晓芳¹，　王永魁²，　邱小忠²，　王国保³

1.遵义医学院珠海校区人体解剖与组织胚胎学教研室，广东珠海 519041； 2.南方医科大学临床解剖学研究所
广东省组织构建与检测重点实验室，广州 510515； 3.南方医科大学南方医院肾内科，广州 510515

收稿日期:2014-03-05 出版日期:2014-09-25 发布日期:2014-10-14
通讯作者: 王国保，副主任医师，副教授，E-mail：Wgbandyl @ medmail.com.cn E-mail:xiaofang4231@163.com
作者简介:胡晓芳（1986- ）女，安徽安庆人，助教，硕士，研究方向：肿瘤的基因治疗
基金资助:
贵州省科学技术基金（黔科合J字[2014]2179号）

Study of the effect of Nix and miR-520h on glioma

HU Xiao-fang¹, WANG Yong-kui², QIU Xiao-zhong², WANG Guo-bao³

1.Department of Anatomy, Zhuhai school Compus of Zunyi Medical College, Zhuhai 519041, China; 2.Department of Anatomy，Key Laboratory of Construction and Detection of Guangdong Province，Southern Medical University，Guangzhou 510515，China；3.Department of Nephrology, Nanfang Hospital，Southern Medical University，Guangzhou 510515，China

Received:2014-03-05 Online:2014-09-25 Published:2014-10-14

摘要/Abstract

摘要：

目的　通过综合分析肿瘤标本中基因和蛋白表达，及体外生物学作用的检测，来评价胶质瘤中Nix蛋白的作用以及Nix蛋白与miR-520h的关系。方法　选取46例胶质瘤标本，RT-PCR检测miR-520h基因的表达，Western blotting检测Nix 蛋白的表达；培养U251胶质瘤细胞，Western blotting检测Nix蛋白的表达；构建靶向Nix基因的shRNA敲除U251细胞中Nix基因（Nix-kn）, Western blotting检测Nix, IKKα, i-κBα和p-NF-κB/p65 等蛋白的表达水平; has-miR-520h抑制剂转染U251细胞，Western blotting检测Nix, IKKα, i-κBα和p-NF-κB/p65 等蛋白的表达水平。结果　胶质瘤标本的基因和蛋白检测表明，miR-520h的高表达伴随着Nix蛋白的高表达；体外实验结果表明，与正常培养的U251相比，缺氧培养的U251细胞Nix表达较高；与Nix-kn组相比，Nix-wt组胶质瘤细胞Nix和p-NF-κB/p65表达均明显增高，而NF-κB活性抑制剂蛋白i-κBα表达降低；200 nmol/L的has-miR-520h抑制剂处理细胞后，与正常对照组比较， Nix蛋白表达显著减少，而p-NF-kappaB/p65蛋白表达显著增加。结论　在神经胶质瘤的发生发展中miR-520h与Nix蛋白的表达相关；miR-520h可能通过促进Nix合成，从而成为一个强大的肿瘤激活剂。

关键词: 胶质瘤, Nix, microRNA-520h, NF-κB

Abstract:

Objective　To evaluate the biological function of Nix and its relation to microRNA miR-520h in glioma.　Methods　46 glioma specimens were selected to detect the expression of Nix by Western blotting and the microRNA expression by traditional RT-PCR．The shRNA were used to knockdown Nix of U251 glioblastomacell line, and the protein level of Nix was detected by Western blotting. In the presense of miR-520h inhibitor．The protein level of Nix and NF-κB was detected to evaluate their interaction in the U25l cell.　Results　Both RT-PCR analysis and Western blotting showed that high expression of miR-520h was always accompanied with high expression of Nix proteins. Under hypoxic conditions, Western blotting showed that levels of both Nix and p-NF-κB/p65 in Nix-wt U251 glioblastoma cells were significantly higher than those in Nix-kn U251 glioblastoma cells. Furthermore，200 nmol/L of has-miR-520h inhibitor caused low expression of Nix and high expression of p-NF-κB/p65 under hypoxic conditions.　Conclusion　MiR-520h is related to the expression of Nix protein in the tumorigenesis of glioma. MiR-520h may be a strong tumor activator through promoting Nix biosynthesis .

Key words: Glioma, Nix, miR-520h, NF-κB

中图分类号:

R739.41

胡晓芳, 王永魁, 邱小忠, 王国保. Nix和miR-520h对神经胶质瘤发生的影响及相互作用[J]. 中国临床解剖学杂志, 2014, 32(5): 590-594.

HU Xiao-Fang, WANG Yong-Kuai, QIU Xiao-Zhong, WANG Guo-Bao. Study of the effect of Nix and miR-520h on glioma[J]. Chinese Journal of Clinical Anatomy, 2014, 32(5): 590-594.

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Nix和miR-520h对神经胶质瘤发生的影响及相互作用

Study of the effect of Nix and miR-520h on glioma

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