组蛋白去乙酰化酶抑制剂RGFP109逆转胶质母细胞瘤U251细胞对替莫唑胺耐药性的机制研究

发布时间：2018-03-20 12:09

本文选题：组蛋白去乙酰化酶抑制剂RGFP　切入点：转录因子κB　出处：《中国药房》2017年22期 　论文类型：期刊论文

【摘要】：目的:研究组蛋白去乙酰化酶抑制剂RGFP109逆转胶质母细胞瘤U251细胞的耐药性机制。方法:建立对替莫唑胺(TMZ)耐药的TR/U251细胞,试验分为正常对照组、TMZ组(40μmol/L)和TMZ(40μmol/L)+RGFP109(0~120μmol/L)不同浓度组,加入相应药物作用24 h后,CCK-8法检测细胞存活率,计算半数抑制浓度(IC_(50))。TUNEL法和Annexin V/PI法检测正常对照组、TMZ组和TMZ+RGFP109(42μmol/L)组细胞的凋亡情况,免疫印迹法检测3组细胞中O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)、存活蛋白(Survivin)、B淋巴细胞瘤2(Bcl-2)、B淋巴细胞瘤x L(Bcl-x L)蛋白表达,凝胶迁移实验检测3组细胞中p65乙酰化水平及其与κB-DNA的结合能力。结果:与正常对照组和TMZ组比较,TMZ+RGFP109不同浓度组细胞存活率明显降低(P0.05),且呈浓度依赖性。当RGFP109浓度为42μmol/L时,TMZ对TR/U251细胞的敏感性与U251细胞相同。与正常对照组比较,TMZ组细胞中MGMT、Survivin、Bcl-2、Bcl-x L蛋白表达均增强(P0.01),p65乙酰化水平无明显变化,但p65与κB-DNA的结合能力增强(P0.01)。与TMZ组比较,TMZ+RGFP109组细胞中MGMT、Survivin、Bcl-2、Bcl-x L蛋白表达均减弱(P0.01),p65乙酰化水平增强(P0.01),p65与κB-DNA的结合能力减弱(P0.01)。结论:RGFP109可通过下调转录因子κB调节的抗凋亡蛋白表达,减弱p65与κB-DNA的结合来逆转U251细胞对TMZ耐药。
[Abstract]:Aim: to study the mechanism of protein-deacetylase inhibitor (RGFP109) in reversing drug resistance of glioblastoma U251 cells. Methods: TR/U251 cells resistant to temozolidomide were established and divided into control group (40 渭 mol / L) and TMZ(40 渭 mol / L (RGFP109(0~120 渭 mol / L) groups. The cell survival rate was detected by CCK-8 method after 24 h treatment with corresponding drugs. The apoptosis of normal control group and TMZ RGFP109(42 渭 mol / L group was detected by Tunel method and Annexin V / Pi method. The expression of O6-methylguanine-DNA methyltransferase (MGMTA) and survivin B lymphocytoma (Bcl-2C ~ (2 +) B lymphocytoma x Bcl-x L) was detected by Western blotting. The level of p65 acetylation and its binding ability to 魏 B-DNA were detected by gel migration assay. Results: compared with normal control group and TMZ group, the cell survival rate of different concentrations of TMZ RGFP109 decreased significantly in a dose-dependent manner. The sensitivity of TR/U251 cells to TR/U251 cells was the same as that of U251 cells at the concentration of 42 渭 mol/L. However, the binding ability of p65 to 魏 B-DNA was enhanced and p0.01. Compared with TMZ group, the expression of MGMT-survivin Bcl-2Bcl-xL protein in TMZ RGFP109 group was decreased. The acetylation level of p0.01p65 increased the binding ability of p0.01p65 to 魏 B-DNA. Conclusion: RGFP109 can down-regulate the expression of anti-apoptotic protein regulated by transcription factor B. The combination of p65 and 魏 B-DNA was reduced to reverse the drug resistance of U251 cells to TMZ.
【作者单位】：中山大学附属第八医院药学部;深圳大学附属第一医院神经外科;
【分类号】：R96

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