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抑制组蛋白去乙酰化酶与DNA甲基转移酶活性对胶质瘤恶性表型影响的研究

王晓蕊1,杜汋2,浦佩玉3,康春生4   

  1. 1. 天津市肿瘤医院
    2. 天津市卫生局
    3. 天津医科大学总医院
    4. 天津医科大学总医院神经外
  • 收稿日期:2012-02-29 修回日期:2012-08-07 出版日期:2012-11-15 发布日期:2012-11-15
  • 通讯作者: 王晓蕊

Study on the suppressive effect of histone deacetylase and DNA methyltransferase inhibitors on malignant phenotypic characteristics of human glioma cells

  • Received:2012-02-29 Revised:2012-08-07 Published:2012-11-15 Online:2012-11-15
  • Contact: Xiao-rui WANG

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摘要： 【摘要】 目的：探讨阻断组蛋白去乙酰化酶与DNA甲基转移酶的活性后对胶质瘤恶性表型的抑制作用。方法：选择丙戊酸（VPA）为组蛋白去乙酰化酶抑制剂，5-氮杂-2′-脱氧胞苷（Aza）为DNA甲基转移酶抑制剂。将U251胶质瘤细胞分为对照组、VPA治疗组、Aza治疗组和VPA+Aza联合治疗组。采用四唑盐(MTT)比色法分析肿瘤细胞增殖活性，流式细胞术分析细胞周期，Annexin V-FITC染色检测细胞凋亡，2D Matrigel、3D Matrigel、Transwell法检测胶质瘤细胞侵袭能力，并建立U251细胞裸鼠皮下移植瘤模型，进行肿瘤局部多点注射上述药物治疗，测量肿瘤体积。结果：与对照组比较，各治疗组肿瘤细胞的增殖在治疗2 d后均出现明显抑制，S期和G2M期细胞比例减少，诱导细胞周期阻滞于G0/G1期，治疗组细胞凋亡率明显下降，侵袭和运动迁移能力均明显下降，各治疗组鼠移植瘤体积增长较对照组明显减缓，与对照组相比差异均有统计学意义（P<0.05）。以上结果均以VPA+Aza联合治疗组最为显著。结论：联合阻断DNA甲基转移酶和组蛋白去乙酰化酶活性可抑制胶质瘤表观遗传学性征，抑制胶质瘤恶性表型。

关键词: 神经胶质瘤, 组蛋白去乙酰化酶抑制剂, DNA修饰甲基酶类, 细胞增殖, 肿瘤侵润, 细胞凋亡

Abstract: 【Abstract】 Objective To investigate the suppressive effect of histone deacetylase and DNA methyltransferase inhibitors on the malignant phenotype of human glioma cells by impacting on the epigenetic mechanism.Methods VPA (Valproic acid) was used as histone deacetylase inhibitor and Aza (5-aza-deoxycytidine) as DNA methyltransferase inhibitor. For in vitro study, U251 glioma cells were divided into four groups: the control group, VPA treatment group, Aza treatment group and the VPA+Aza combined treatment group. The proliferation activity of tumor cells was detected by MTT assay, cell cycle analysis by flow cytometry, apoptosis rate by Annexin V-FITC assay and the invasion ability of glioma cells by 2D Matrigel, 3D Matrigel, and transwell assay. The subcutaneous U251 tumor model was established in nude mice for in vivo study, xenografts were treated with VPA, Aza and VPA+Aza by multi-point local injection into tumor bed every 4 days within the observation period for 28 days and the gross tumor volume was measured. Results Compared with the control group, the tumor cell proliferation activity in VPA , Aza and VPA + Aza combined treatment group, was significantly lowered(P <0.05 for each group), the cells were arrested in G0/G1 phase, apoptotic cell rate was increased and cell invasion ability decreased significantly (P <0.05). The tumor volume in all treatment groups was much smaller than that in control group. The most significant effect on inhibition of malignant biological behaviour of glioma cells was observed in VPA + Aza combined treatment group. Conclusion To treat glioma cells with DNA methyltransferase and histone deacetylase inhibitors can reverse the malignant phenotype by inhibition of their epigenetic alterations, So this study provides the evidence that both of them can be developed as candidates for glioma treatment.

Key words: Glioma, histone deacetylase inhibitor, DNA methyltransferase inhibitor, Proliferation, Invasion, Apoptosis