摘要
目的分离前列腺癌雄激素依赖型和非依赖型细胞系差异表达蛋白质,为进一步研究前列腺癌的蛋白组学打下基础。方法培养前列腺癌雄激素依赖型(LNCaP)和非依赖型(DU145)细胞系,分别提取两种细胞系的总蛋白,进行等电点聚焦和SDS-聚丙烯酰胺凝胶电泳,对所得胶图利用ImageMaster4.01软件进行分析,选取差异点进行质谱分析,对分析结果进行数据库检索。结果得到了分辨率和重复性均好的前列腺癌雄激素依赖型和非依赖型细胞系的双向凝胶电泳图谱,每块凝胶上平均蛋白质点数为816±15,通过分析得到3倍差异点41个,完全差异点10个。对完全差异点进行质谱分析和数据库检索得到与差异点相对应的3个蛋白质,其中第83号和175号为未命名蛋白质,第632号蛋白质是KIAA1283蛋白质,具有载脂蛋白功能。结论在前列腺癌激素依赖型和激素非依赖型细胞系中存在差异表达蛋白质,并可以利用双向凝胶电泳技术进行分离,为进一步研究前列腺癌的蛋白质组学和发病机制提供了线索。
Objective To sieve the differential protein from androgen-dependent and androgen-independent cells of human prostate cancer using two-dimensional polyacrylamide gel electrophoresis（2D-PAGE） with matrix-assisted laser desorption and ionization-time of flight mass spectrometry（ MALDI-TOF MS）. Methods These two different cell lines were cultured, and total proteins were extracted from these cells. After preparation of the total proteins, solubilized proteins were separated in the first dimension in IPG（immobilized pH gradient） strips depending on their pI and subsequently in the second dimension according to their molecular weight by SDS-polyacrylamide gel electrophoresis. Those differential protein spots were visualized by image analysis software, and several completely differential spots were isolated from the gels and identified by tryptic cleavage and subsequent MALDI-TOF mass spectrometry. Results The repeatability and resolution of Gel electrophoresis were high. Average protein spots were 816± 15. More than 50 differential protein spots were visualized ,of which, 10 completely differential spots were isolated from the gels and identified by MAL- DI-TOF mass spectrometry. Among them No. 83 and No. 175 protein were not reported before,and No. 632 protein was reported as KIAA1283 protein, a kind of apolipoprotein. Conclusion The results indicate that there are differential proteins between androgen-dependent and androgen-independent cells of human prostate cancer,and further research of the molecular pathogenesis of prostate cancer can be based on it.
出处
《山西医科大学学报》
CAS
2008年第4期293-297,385,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(30371420)
关键词
前列腺肿瘤
双向凝胶电泳
质谱
prostate neoplasms
two-dimensional polyacrylamide gel electrophoresis
mass spectrum
