论文部分内容阅读
目的:建立利用二维液相色谱法分离肿瘤细胞全细胞裂解液的分析方法,进而研究金线莲水提物的抗肿瘤作用以及肿瘤细胞蛋白质组的差异。方法:将金线莲水提物给药后的肿瘤细胞裂解样品及对照样品用初始缓冲液置换后,进行一维色谱聚焦分离,然后先对二维色谱条件进行优化和重复性分析,再将一维收集的pH4.0～8.5之间的组分分别进行二维无孔硅胶HPLC分离,利用ProteoVue软件将UV图转换成胶图,分析差异。结果:一维色谱聚焦分离pH4.0～8.5之间组分共收集到16个,每个组分的二维UV图转换成PI/UV胶图,结果表明金线莲水提物给药前后肿瘤细胞蛋白质组差异具显著性。结论:二维液相色谱分离法是一种有效的分离肿瘤细胞裂解液的方法,给药后的肿瘤细胞蛋白质组和对照相比差异具显著性。
OBJECTIVE: To establish an analytical method for separating whole cell lysates of tumor cells by two-dimensional liquid chromatography, and to study the antitumor effects of the aqueous extract of Anoectochilus roxburghii and the proteome differences of tumor cells. Methods: Tissue samples of lysed tumor cells after the extract of Anemarrhena astigmatism and control samples were replaced by initial buffer, and then one-dimensional chromatographic separation was carried out. Then the two-dimensional chromatographic conditions were optimized and repeatability. Two-dimensional non-porous silica gel HPLC separations were collected from one-dimensional fractions of pH 4.0 to 8.5, and the UV maps were converted to gel using ProteoVue software to analyze the differences. Results: One-dimensional chromatographic separation of pH4.0 ~ 8.5 between the components collected a total of 16, each component of the two-dimensional UV map converted to PI / UV glue map, the results show that the extract of Anoectochilus before and after The difference of tumor cell proteome was significant. Conclusion: Two-dimensional liquid chromatography is an effective method for separating tumor cell lysate. The difference of the proteome of tumor cells after administration is significant compared with the control.