γ-羟丁酸钠对人胶质瘤细胞SHG-44放疗增敏的实验研究

发布时间：2018-02-13 00:06

本文关键词： 胶质瘤 γ-羟丁酸钠放疗增敏　出处：《苏州大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：探讨γ-羟丁酸钠（gamma-hydroxybutrate，GHB）对胶质瘤细胞SHG-44增殖抑制及放疗增敏机制研究。方法：以SHG-44细胞为研究对象，MTT法检测不同浓度GHB（2.5mmol/L、5mmol/L、7.5mmol/L、10mmol/L、15mmol/L）作用人胶质瘤细胞不同时间（24h、48h、72h）细胞增殖抑制率。Hoechst33258实验观察不同浓度GHB（5mmol/L、10mmol/L、15mmol/L）作用24h后细胞核变化。下列实验分为对照组、药物组、照射组、联合组（照射+药物）。流式细胞术（FCM）分析各组细胞周期、凋亡情况。克隆形成实验分析GHB联合不同剂量的X线照射对细胞存活率的影响，拟合存活曲线。细胞免疫荧光检测SHG-44细胞中HDAC1（Histone deacetylase1，HDAC1）mRNA表达情况。RT-PCR检测各组细胞HDAC1mRNA、ATM （Ataxiatelangiectasia-mutated gene）mRNA的转录水平。结果：MTT实验显示GHB明显抑制SHG-44细胞的生长，呈浓度和时间依赖性。Hoechst33258镜下观察时染色体浓缩，见强蓝色荧光。FCM检测联合组细胞周期S期细胞减少，G0/G1期细胞增多，凋亡显著。克隆形成实验显示联合组的D0、Dq及SF2各值低于照射组(1.764Vs2.237，，1.251Vs1.873，0.561Vs0.769)，联合组SER为1.27。细胞免疫荧光测得HDAC1主要在SHG-44细胞核中表达。RT-PCR结果表明γ-羟丁酸钠能抑制HDAC1mRNA、ATM mRNA表达。结论：GHB通过诱导细胞凋亡，改变细胞周期，抑制肿瘤细胞损伤修复，抑制HDAC1、ATM mRNA表达，从而增强胶质瘤细胞放射敏感性。
[Abstract]:Aim: to investigate the mechanism of 纬-hydroxybutyrate sodium gamma-hydroxybutyrate (GHBs) inhibiting the proliferation of glioma cell line SHG-44 and enhancing the sensitivity of radiotherapy. Methods: the proliferation inhibition rate of human glioma cells treated with SHG-44 cells at different concentrations of 2.5 mmol 路L ~ (2.5) mmol 路L ~ (5) mmol 路L ~ (5) mmol 路L ~ (5) mmol 路L ~ (10) mmol 路L ~ (10) mmol 路L ~ (10) mmol 路L ~ (10) mmol 路L ~ (15) mmol 路L ~ (-1) 路Hoechst33258 (24 h / 24 h) was measured by MTT assay. The following experiments were divided into two groups: control group, drug group, 10 mmol 路L ~ (10) mol 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L). In the irradiation group, the cell cycle and apoptosis were analyzed by flow cytometry (FCM). Clone formation assay was used to analyze the effect of GHB combined with different doses of X-ray irradiation on cell survival. Cell immunofluorescence was used to detect the expression of HDAC1(Histone deacetylase1HDAC1mRNA in SHG-44 cells. RT-PCR was used to detect the transcription level of HDAC1mRNA-Ataxiatelangiectasia-mutated gene)mRNA. Results GHB significantly inhibited the growth of SHG-44 cells in a dose-and time-dependent manner. Chromosome concentration was observed in a dose-and time-dependent manner. Strong blue fluorescence. FCM assay showed that the number of G _ 0 / G _ 1 phase cells decreased in the S phase of cell cycle. The results showed that D0 Dq and SF2 in the combined group were lower than those in the irradiated group (1.764 vs 2.237) and 1.871 vs 1.871 vs 0.561Vs0.769in the combined group. The results of cellular immunofluorescence assay showed that HDAC1 mainly expressed in the nucleus of SHG-44. The results of RT-PCR showed that 纬 -hydroxybutyrate could inhibit the expression of HDAC1mRNAs mRNA. Conclusion by inducing apoptosis, changing cell cycle, inhibiting the repair of tumor cell injury and inhibiting the expression of HDAC1 mRNA, the radiosensitivity of glioma cells can be enhanced.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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