摘要
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 develop an approach for fractionating cancer cell lysates by two-dimensional （2-D） liquid phase chromatographic fractionation, then to study the anti-tumor effect of aqueous extract of Anoectochilus roxburghii, and to analyze the difference of the proteome of the tumor cell. METttODS The cancer cell lysates were exchanged with start buffer and separated by chromato-focusing in the first dimension, which were followed by optimizing the chromatographic condition of the second dimension and analysis of reproducibility, the fractions between pH 4. 0 and pH 8. 5 were separated by non-porous silica reverse phase HPLC. The UV maps were transformed into gel-like maps by ProteoVue software. RESULTS There were 16 fractions between pH 4. 0 and pH 8. 5 after chromato-focusing in the first dimension and the UV maps of each fraction were transformed into PI/UV gel-like maps,and it showed distinct changes between drug-treated and untreated cancer cells in proteome. CONCLUSION 2-D phase chromatographic fractionation is an effective method to separate cancer cell lysates. The proteome of the tumor cell after drug-treatment has substantial difference with the control group.
出处
《中国医院药学杂志》
CAS
CSCD
北大核心
2011年第8期623-626,共4页
Chinese Journal of Hospital Pharmacy
基金
福建省科技厅重点资助项目(编号:2009I0009)
关键词
金线莲水提物
蛋白质组
肿瘤细胞
二维液相色谱分离
Anoectochilus roxburghii
proteome
cancer cell
two dimensional liquid phase chromatographic fractionation
