白桦脂酸增强抗肿瘤药物诱导U266细胞凋亡及其机制研究

发布时间：2018-04-24 11:01

本文选题：白桦脂酸 + 抗肿瘤药物　；参考：《贵阳医学院》2015年硕士论文

【摘要】：目的:探讨白桦脂酸增强抗肿瘤药物诱导U266细胞凋亡及其机制研究。方法:用不同浓度的白桦脂酸(20μg/m L,40μg/m L,60μg/m L,80μg/m L)、硼替佐米(10 nmol/L,80 nmol/L,320 nmol/L),沙利度胺(10μg/m L,50μg/m L,100μg/m L)和马法兰(5μmol/L,10μmol/L,20μmol/L)及白桦脂酸联合不同浓度的硼替佐米,沙利度胺及马法兰分别处理U266细胞,MTT法检测细胞增殖抑制率,Annexin V/PI法流式细胞仪检测不同浓度硼替佐米、沙利度胺、马法兰单独用药及分别与白桦脂酸联合用药的各组细胞凋亡率;Real-time PCR检测Survivin,Cyto-C,Bcl-2,Bax基因表达水平;蛋白免疫印迹法检测Survivin,Cyto-C,Bcl-2,Bax蛋白表达水平。结果:联合白桦脂酸用药组的U266细胞对硼替佐米、沙利度胺及马法兰的增殖抑制率显著高于单用药组(p0.05);流式细胞术结果显示:白桦脂酸与硼替佐米联合用药处理多发性骨髓瘤U266细胞后,10 nmol/L,80nmol/L,320 nmol/L细胞凋亡率分别为(36.94±0.04)%、(56.73±0.08)%、(78.67±0.03)%;同样的白桦脂酸和沙利度胺(10μg/m L,50μg/m L,100μg/m L)联合用药后各组的凋亡率分别为(27.32±0.02)%、(46.16±0.03)%、(70.17±0.08)%;白桦脂酸和马法兰(5μmol/L,10μmol/L,20μmol/L)联合用药后各组的凋亡率分别为(37.43±0.07)%、(51.78±0.02)%、(74.19±0.05)%,联合用药组合单用药组比较均有显著性差异(P0.05);RT-PCR和Western Blot结果显示:联合白桦脂酸用药处理多发性骨髓瘤U266细胞后Survivin,Bcl-2 m RNA和蛋白表达水平显著降低(p0.05);而Cyto-c和Bax表达水平显著升高(p0.05)。结论:白桦脂酸联合硼替佐米、沙利度胺及马法兰可以诱导U266细胞凋亡,其机制可能与联合用药后下调Survivin,Bcl-2,上调cyto-c、Bax表达相关。
[Abstract]:Objective: to investigate the mechanism of U266 cell apoptosis induced by antitumor drugs enhanced by Bai Hua. Methods: different concentrations of Bai Hua, 20 渭 g / mL, 40 渭 g / L, 60 渭 g / L, 80 渭 g / L, 80 nmol / L, 80 nmol / L, 10 nmol / L, 10 渭 g / L, 10 渭 g / m / L, 50 渭 g / m / L, 50 渭 g / m / L, 100 渭 g / L / L), 10 渭 mol / L / L ~ 10 渭 mol / L ~ (10 渭 mol / L ~ (20 渭 mol / L), 10 渭 mol / L ~ (20 渭 mol / L), 10 渭 mol / L, 10 渭 mol / L ~ (20 渭 mol / L), 10 渭 mol / L, 10 渭 mol / L ~ (10 渭 mol / L), 10 渭 mol / L ~ (20 渭 mol / L)), Thalidomide and Mavan treated U266 cells were used to detect the inhibition rate of cell proliferation by MTT assay. Flow cytometry was used to detect bortezomide and thalidomide at different concentrations by Annexin V/PI. The expression of survivin Cyto-Con Bcl-2Bcl-2Bcl-2Bax gene was detected by real-time PCR, and the expression of survivvin-Cyto-Con Bcl-2Bcl-2Bcl-2Bax protein was detected by Western blotting. Results: U266 cells in combination with Bai Hua group were treated with bortezomib. The proliferation inhibition rate of thalidomide and manfarin was significantly higher than that of control group (P 0.05), and the results of flow cytometry showed that the apoptotic rate of 10 nmol / L ~ (80) nmol / L ~ (320) nmol/L cells in multiple myeloma U266 cells treated with Bai Hua and bortezomil was 36.94 卤0.044.73 卤0.088.67 卤0.03, respectively. The apoptotic rate of each group was 27.32 卤0.02 卤0.02 and 70.17 卤0.08 respectively after the same combination of Bai Hua and 10 渭 g / mL of thalidomide, and the apoptotic rate of the three groups was 37.43 卤0.077.78 卤0.024.19 卤0.055.The apoptotic rate of the three groups was 37.43 卤0.077.78 卤0.02g / ml, respectively, and the apoptotic rate of the two groups was 37.43 卤0.077.78 卤0.024.19 卤0.055.The apoptotic rate of the three groups was 37.43 卤0.077.78 卤0.02 卤0.054.19 卤0.055.The apoptotic rate of the two groups was 37.43 卤0.071.78 卤0.024.19 卤0.055.The apoptotic rate of each group was 37.43 卤0.071.78 卤0.024.19 卤0.055.The apoptotic rate of the two groups was 37.43 卤0.071.78 卤0.024.19 卤0.05. The results of RT-PCR and Western Blot showed that the expression of survivin Bcl-2 m RNA and protein in U266 cells treated with Bai Hua was significantly decreased, while the expression of Cyto-c and Bax increased significantly. Conclusion: Bai Hua combined with bortezomil, thalidomide and manfarin can induce apoptosis of U266 cells. The mechanism may be related to the down-regulation of survivin Bcl-2 and up-regulation of cyto-cor Bax expression.
【学位授予单位】：贵阳医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R96

【引证文献】