DADS诱导人结肠癌细胞双向电泳图谱的差异分析

目的近年来,蛋白质组学技术已广泛应用于肿瘤研究领域,但对结肠癌的研究较少。该实验旨在研究二烯丙基二硫(DADS)体外诱导人结肠癌SW 480细胞蛋白质组差异表达的相关分子机制。方法采用蛋白质组学相关技术,如二维电泳、图像分析、质谱技术等方法,获得DADS体外培养的人结肠癌SW 480细胞蛋白质组的肽质量指纹图,将所得的数据进行生物信息学处理。结果在相同条件下,对DADS处理前后SW 480细胞两种样品的总蛋白质进行双向电泳,经PDQuest软件分析,结果显示,其中167个匹配的蛋白质点存在表达量上的差异,与对照组相比,处理组蛋白质表达量下调的有69个,利用基质辅助激光解吸/电离飞行时间质谱技术(MALD I-TOF-MS)对其中8个表达明显下调的蛋白质斑点进行分析鉴定,获得相应的肽质量指纹图谱(PMF),通过数据库搜索,初步确定这8个蛋白质分别是:细胞角蛋白19(Cytokeratin-19)、肌动蛋白解聚因子(Actin-de-polym erizing factor)、V-1蛋白(V-1 prote in)、果糖二磷酸醛缩酶(Fructose b isphosphate aldolase,FBA)、FK506结合蛋白(FK506-b ind ing prote in,FKBP)、成帽蛋白(Capp ing prote in)、硫氧还蛋白过氧化物酶(Th ioredoxin peroxidase)和敏感性转化蛋白(Transform ation-sensitive prote in)。结论这些差异蛋白质涉及细胞周期调控、细胞分化、细胞凋亡及细胞分裂增殖等众多事件,从而说明,DADS可能具有抑制结肠癌细胞生长,阻滞细胞周期,诱导细胞分化及凋亡的作用。

The differential proteomic expression analysis of diallyl disulfide-induced human colonic cancer cells

Aim This study was designed to explore the differential proteomic expression inducing effect of diallyl disulfide(DADS) on human colonic cancer SW480 cells and its related molecular mechanisms.Methods A series of methods,including immobilized pH gradient-two dimensional polyacrylamide gel electrophoresis,peptide mass fingerpringting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS),database searching and so on,were used to separate and identify the differential proteomic expressions inducing effect of DADS on human colonic cancer SW480 cells.Results The results showed that the good 2-DE pattern including high resolution and reproducibility was obtained.After silver staining,the 2-DE image was analyzed with PDQuest 2-DE software.There were 167 spots that exhibited detectable quantitative changes between control and DADS-treated cells,69 of them realized down-regulation.The expression of cytokeratin-19,actin-depolymerizing factor,V-1 protein,fructose bisphosphate aldolase,FK506-binding protein,capping protein,thioredoxin peroxidase and transformation-sensitive protein were down-regulated in DADS-treated cells.Conclusion There are differential expression proteins between control and DADS-treated cells.Searching differential expression proteins between control and DADS-treated cells provides insight into the changed global protein patterns of DADS-treated cells and clues for further investigation of the molecular mechanism of colonic cancer.