组蛋白去乙酰化酶抑制剂曲古抑菌素A对肝癌细胞蛋白质差异表达谱的影响

Effects fo Trichostatin A on the differentially expressed profile of proteins in hepatocelluar carcinoma

目的观察组蛋白去乙酰化酶（HDAC）抑制剂-曲古抑菌素A（TSA）对肝癌细胞（BEL-7402）蛋白质表达谱的影响。方法采用150g／LTSA处理BEL-7402细胞16hr为试验组，未处理的细胞为对照组，分别抽提其总蛋白质，蛋白定量恒定为200mg。经第一向固相pH梯度等电聚焦电泳和第二向垂直平板SDS—PAGE，凝胶银染后获得二维凝胶图并扫描输入计算机进行软件分析。将三次重复实验所获得的二维凝胶图进行统计学分析。将差异表达的蛋白质斑点从银染后的二维凝胶中切下，进行质谱分析。结果实验组BEL-7402细胞90％～95％的蛋白质表达与对照组一致，5％-10％的蛋白质为差异表达；通过质谱对差异点的鉴定，获得了16个差异表达蛋自质，包括：Triosephosphate isomerase．47-KDa heat shock protein、TRAF1，5（up）、annexin A1、Heat shock 70 protein、prohibitin、thioredoxin peroxidase、DP-1等分子，这些蛋白质涉及基因表达调控、信号转导、细胞代谢、抑制细胞增殖等众多分子事件。结论TSA能够通过影响蛋白质的差异表达影响BEL-7402肝癌细胞的生长和增殖。

Objective To study the effects of Trichostatin A （TSA） on the differentially expressed profile of proteins in hepatocelluar carcinoma （HCC）. Methods Proteomic technique and microarray were performed to analyze the differentially expressed profile in HCC cell line BEL-7402HCC after treated with 150 mg/ml TSA for 16 h. The 2-D and MS were performed to analyze the effect of TSA on protein expression profile in HCC cells. Results There were 5% -10% differentially expressed proteins. Eight up-regulated proteins and eight down-regulated proteins in the transfected cells were successfully identified by ESI-Q-TOF. These differentially expressed proteins included： riosephosphate isomerase, 47-kD heat shock protein,TRAF1,5 （ up）, annexin A1, heat shock 70 protein, prohibitin, thioredoxin peroxidase, DP- 1, and so on. vThese differentially expressed proteins involved cell cycling, transcription regulation, signaling pathway, etc. Conclusion TSA may exert its effect by mediating differential expression of these proteins.