新型组蛋白去乙酰化酶抑制剂Fa对T细胞淋巴瘤细胞株Jurkat的抗肿瘤机制研究

发布时间：2018-04-24 22:29

本文选题：组蛋白去乙酰化酶抑制剂 + T细胞淋巴瘤　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:研究新型组蛋白去乙酰化酶抑制剂Fa的抗肿瘤活性作用及其抑制人T细胞淋巴瘤细胞的增殖机制。方法:1、通过CCK-8法检测不同浓度Fa(1.56,3.125,6.25,12.5,25,50μM)对Jurkat细胞作用24、48、72h的增殖抑制作用,并计算出三个时间段细胞增殖抑制半数的药物浓度(IC50);2、流式细胞术观察不同浓度Fa对Jurkat细胞作用72h后诱导细胞周期阻滞的作用以及25μM Fa作用细胞24、48、72h后细胞周期变化;3、Western blot测定Fa对Jurkat细胞中cyclin D、CDK4、p21~(cip/WAF)的蛋白表达影响;4、RT-PCR测定Fa对Jurkat细胞中HDAC1、HDAC2、HDAC3基因的表达影响;5、实验数据以均数±标准差表示,应用Prism 5软件进行统计分析,采用单因素方差分析(One-way ANOVA)及两样本均数比较的t检验,P0.05表示差异具有统计学意义。结果:CCK-8检测结果显示,以正常细胞作对照,不同浓度Fa作用Jurkat细胞24、48、72h后,细胞增殖抑制半数的药物浓度(IC50)分别是88.72±0.13、25.45±0.03、12.21±0.07μM,作用72h时,细胞生长活力百分比由95.6±2.57%减少至11.4±1.41%,并且呈浓度和时间依赖性;流式细胞术分析结果显示不同浓度Fa作用Jurkat细胞72h后可以产生G0/G1期细胞周期阻滞,并且呈浓度依赖性。25μM Fa作用时,随着时间延长,处于G0/G1期细胞百分比由42.54±2.11%增加至61.42±0.59%;p21~(cip/WAF)蛋白表达水平有所上升,cyclin D、CDK4蛋白表达下调;Fa药物能够有效抑制HDAC1、HDAC2、HDAC3的活性。结论:Fa对T细胞淋巴瘤细胞株具有一定的抗肿瘤活性,其机制与诱导细胞凋亡周期阻滞及上调抑癌基因p21~(cip/WAF)表达有关。
[Abstract]:Aim: to study the antitumor activity of a novel histone deacetylase inhibitor Fa and its mechanism of inhibiting the proliferation of human T cell lymphoma cells. Methods CCK-8 assay was used to detect the proliferation inhibitory effect of different concentrations of Fa 1.56C 3.125C 6.255C 2550 渭 M on Jurkat cells for 72 h. The cell cycle arrest induced by different concentration of Fa for 72 h and the cell cycle change after treatment with 25 渭 M Fa for 72 h were observed by flow cytometry. Effect of Fa on the protein expression of cyclin DCDK4p21cip-pWAF in Jurkat cells by Western blot the effect of Fa on the expression of HDAC1HDAC2HDAC3 gene in Jurkat cells was determined by RT-PCR. The experimental data were expressed as mean 卤standard deviation. The statistical analysis was carried out by using Prism 5 software. One-way ANOVA) and the t test of the mean of the two samples were used to show the difference was statistically significant. Results the cell proliferation inhibition 50 (IC50) was 88.72 卤0.1325.45 卤0.03C 12.21 卤0.07 渭 M at 72 h after treatment with Fa at different concentrations of Fa. Results the results showed that the IC50 concentration was 88.72 卤0.1325.45 卤0.03n 12.21 卤0.07 渭 M, respectively, in Jurkat cells treated with different concentrations of Fa for 72 h, the results showed that the concentration of IC50 was 88.72 卤0.135.45 卤0.03U 12.21 卤0.07 渭 M. The percentage of cell growth activity decreased from 95.6 卤2.57% to 11.4 卤1.41%, and showed a concentration-and time-dependent manner. Flow cytometry showed that different concentrations of Fa could induce cell cycle arrest in G0/G1 phase after 72 h of treatment with Fa in a concentration-dependent manner. As time went on, the percentage of cells in G0/G1 phase increased from 42.54 卤2.11% to 61.42 卤0.59%. ConclusionTwo Fa has antitumor activity on T cell lymphoma cell lines, and its mechanism is related to the arrest of apoptosis cycle and the up-regulation of the expression of tumor suppressor gene p21cip/ WAF.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.1

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