首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (3): 380-384.doi: 10.3969/j.issn.1006-7795.2013.03.012

• 基础研究 • 上一篇    下一篇

DNA甲基化转移酶3B在脑胶质母细胞瘤中的差异表达

白宇, 陈彦, 国添柱, 刘庆阳, 杨雪, 冯颖, 闫卓红, 王丽佳, 张权庚   

  1. 首都医科大学基础医学院免疫学系,北京 100069
  • 收稿日期:2013-03-04 出版日期:2013-06-21 发布日期:2013-06-17
  • 通讯作者: 张权庚 E-mail:zhangqg@ccmu.edu.cn
  • 基金资助:

    国家自然科学基金(81071626, 30872933)。

Differential expressions of DNMT3B gene in glioblastoma

BAI Yu, CHEN Yan, GUO Tianzhu, LIU Qingyang, YANG Xue, FENG Ying, YAN Zhuohong, WANG Lijia, ZHANG Quangeng   

  1. Department of Immunology, School of Basic medical Science, Capital Medical University, Beijing 100069, China
  • Received:2013-03-04 Online:2013-06-21 Published:2013-06-17
  • Supported by:

    This study was supported by National Natural Science Foundation of China (81071626,30872933).

摘要:

目的 脑胶质母细胞瘤(glioblastoma,GBM)中存在广泛的基因组低甲基化,本研究检查了DNA甲基化转移酶3B (DNA methyltransferase 3B,DMNT3B)是否在这种肿瘤中的表达和突变情况,以期寻找肿瘤基因组低甲基化发生的原因。方法用RNA抽提试剂盒从 25例人脑胶质母细胞瘤组织中提取总RNA,用反转录聚合酶链反应方法扩增 DNMT3B 的cDNA,然后测序,所得序列经DNASTAR软件与NCBI发表的正常序列进行比对。结果 从25例脑胶质母细胞瘤样本中的成功扩增了全长 DNMT3B cDNA,测序分析结果显示 DNMT3B 基因在该肿瘤样本中差异性存在多种剪接变异体,以 DNMT3B-V1、DNMT3B-V3、DNMT3B-V7 为主。没有发现改变氨基酸序列的点突变。结论 DNMT3B 基因在脑胶质母细胞瘤中没有点突变,但存在多种剪接变异体(以 DNMT3B-V3、DNMT3B-V7 为主),其是否为脑胶质母细胞瘤的广泛基因组低甲基化的原因尚有待进一步研究。

关键词: 脑胶质母细胞瘤, 基因组低甲基化, DNA甲基化转移酶3B

Abstract:

Objective Global genomic hypomethylation widely exists in glioblastoma (GBM). This study aimed to clarify the molecular mechanism of this abnormality. Methods Totally 25 GBM samples were examined in this study.Total RNA was extracted from these patient samples. Full-length cDNA was amplified by RT-PCR and sequenced. Sequences obtained from each sample were compared to NCBI published standard sequences by using DNASTAR software. Results We successfully amplified DNMT3B from each patient sample. No point mutation which alters amino acids was identified. But several splicing variants were identified in these patient samples which were differentially exited in the tumor samples. Conclusion No point mutation of DNMT3B was found in GBM patient samples. Two major DNMT3B splice variants ( DNMT3b-V3 and DNMT3b-V7 ) were found differentially exist in these tumor samples. Functions of these variants are subject to further elucidation.

Key words: glioblastoma, genomic hypomethylation, DNA methyltransferase 3B

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