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目的:研究丙二酰黄芪皂苷 I 抗白血病 U937 细胞增殖的差异表达蛋白质组。方法:基于 LC Chip Q-TOF 系统建立和优化了一种非标记蛋白质组比较方法-多肽色谱峰面积。方法学研究表明,该方法操作简单、方便,具有较高的重复性和宽动力学范围。然后,将建立的优化方法组合归一化计算应用于新化合物丙二酰黄芪皂苷 I 抗白血病 U937 细胞增殖的差异蛋白质组表达研究。结果:分析结果表明,2 倍以上表达差异的蛋白质 15 个,其中 6 个蛋白质的表达差异获得了 Western blotting 确证。此外,通过差异蛋白质的相互作用分析,PARP1、Caspase-3 和-9 的 Western blot 检测及 Caspase 抑制剂实验,我们发现 Caspase激活在丙二酰黄芪皂苷 I 抗白血病 U937 细胞增殖效应中起重要作用。结论:通过该优化的非标记蛋白质组比较方法,找到了一些与丙二酰黄芪皂苷 I 抗白血病 U937 细胞增殖密切相关的功能蛋白质,为深入开展其分子机理研究奠定了基础。
OBJECTIVE: To study the differentially expressed proteome of malonyl diacetyl saponin I against leukemia U937 cells. Methods: A non-labeled proteome comparison method was established and optimized based on LC Chip Q-TOF system - peak area of peptide chromatogram. Methodological studies show that the method is simple and convenient to operate, has high repeatability and wide dynamic range. Then, the optimized combination of methods was used to normalize the differential proteomic expression applied to the proliferation of the new compound malonyl-azoglycoside I anti-leukemia U937 cells. Results: The results showed that there were 15 proteins with more than 2-fold difference in expression, of which 6 proteins were confirmed by Western blotting. In addition, through the differential protein interaction analysis, Western blot detection of PARP1, Caspase-3 and -9, and Caspase inhibitor experiments, we found that Caspase activation plays an important role in the proliferation of leukemia U937 cells. CONCLUSION: Some functional proteins closely related to the proliferation of U937 cells induced by malonyl azithoside I are found through the optimized non-labeled proteomic comparison method, which lays the foundation for the further study of its molecular mechanism.