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目的:利用蛋白质组学方法,比较雷公藤甲素作用于大肠癌SW480细胞以获得差异性蛋白质,从蛋白质组学角度评价雷公藤甲素对细胞的综合生物学效应。方法:培养SW480细胞,实验分为4组:DMSO 24h组(A组),DMSO 48h组(B组),TP 20nM 24h组(C组),TP 20nM 48h组(D组),进行双向电泳(2-DE)分离,用PDQuest7.1.0软件包进行差异表达蛋白质组分析,基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF)鉴定蛋白质。结果:获得5个同时满足下面3项条件的差异点:①C组与A组之间存在差异;②D组与B组之间存在差异;③D组与C组之间存在差异。其中3个蛋白经质谱鉴定为同一家族蛋白质:14-3-3蛋白β/α、14-3-3蛋白ε、14-3-3蛋白θ。结论:雷公藤甲素作用于SW480细胞的表达蛋白质组与DMSO培养组比较存在明显的差异;TP抑制大肠癌SW480细胞的药理作用,很有可能是通过14-3-3家族蛋白相互作用使细胞周期发生阻滞,使细胞生长增殖受到抑制。
OBJECTIVE: To compare the effects of triptolide on human colorectal cancer SW480 cells by using proteomics method to obtain differential proteins, and evaluate the comprehensive biological effects of triptolide on cells from the perspective of proteomics. Methods: SW480 cells were cultured and divided into four groups: DMSO 24h group (group A), DMSO 48h group (group B), TP 20nM 24h group (group C) and TP 20nM 48h group (group D) 2-DE). Proteins were differentially expressed using the PDQuest7.1.0 software package, and proteins were identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF). Results: Five different points were found to meet the following three conditions at the same time: ① There was a difference between group C and group A; ② There was a difference between group D and group B; ③ There was a difference between group D and group C. Three of these proteins were identified as the same family of proteins by mass spectrometry: 14-3-3 protein β / α, 14-3-3 protein ε, 14-3-3 protein θ. Conclusion: The expression of triptolide in SW480 cells was significantly different from that in DMSO group. TP inhibits the pharmacological effects of SW480 cells by the interaction of 14-3-3 family of proteins, Cycle block, so that cell growth and proliferation was inhibited.