二烯丙基二硫诱导人胃癌MGC803细胞分化的差异蛋白质分析

目的研究二烯丙基二硫(diallyl disulfide,DADS)诱导人胃癌MGC803细胞分化过程中蛋白质的差异表达。方法用固相pH梯度双向凝胶电泳分离DADS处理前后人胃癌MGC803细胞总蛋白,凝胶经过银染后用PDQuest软件进行分析,差异蛋白质点经胶内原位酶解,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)测定肽质量指纹图(PMF),将所得的数据进行生物信息学处理。结果 DADS处理MGC803细胞前后的总蛋白质的双向电泳银染图谱,经扫描成像及PDQuest软件分析,对照组与处理组蛋白质点与平均匹配率分别为(576±14)个和(583±4)个与76%和70%,421个匹配,200个未被匹配。经相关数据库查询,在初步鉴定的差异蛋白质点中,发现24个与细胞分化、肿瘤转移、细胞凋亡、细胞周期、细胞免疫及代谢等相关的蛋白质,如gastric mucin、nM23、CDC2、uPAR、LIMK、RORα、MHC DR-beta-1 chain、TCR等,这些蛋白质可能在胃癌的发生发展中起着潜在的作用。结论 DADS可能通过多种途径诱导MGC803细胞分化。蛋白质组差异表达的初步分析为进一步研究胃癌分化的相关蛋白质及标记物奠定一定基础。

Proteome analysis of human gastric cancer MGC803 cells induced by diallyl disulfide

Aim To investigate the differential proteo-mic expression in human gastric cancer MGC803 cells induced by diallyl disulfide(DADS).Methods A series of methods,including immobilized pH gradient-two dimensional polyacrylamide gel electrophoresis,silver staining,PDQuest 2-DE software analysis,peptide mass fingerpringting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS) and SWISS-PROT database searching,were used to separate and identify the differential proteomic expressions in MGC803 cells induced by DADS.Results The results showed that the good 2-DE pattern including high resolution and reproducibility was obtained.After silver staining,the 2-DE image analysis by PDQuest 2-DE software detected average spots were(576±14) spots and(583±4) and the matching rate was 76% and 70% in DADS untreated and treated,respectively.The differential proteomic expression analysis found that there were 421 spots matched and 200 spots unmatched between untreated and treated maps.In SWISS-PROT database with MS-FIT software,twenty-four proteins were preliminarily identified related to differentiation,cell cycle,apoptosis,metastasis,cytoimmunity and metabolism,such as gastric mucin,nM23,CDC2,uPAR,LIMK,RORα,MHC DR-beta-1,TCR,etc.There was a significant difference at protein level between untreated and treated MGC803 cells.Conclusions DADS might induce differentiation in human gastric cancer MGC803 cells through different ways.It suggests that the differential expression analysis of proteome may be useful to further study of differentiation related proteins and markers in gastric cancer.