基于差异蛋白质组学解析消痰散结方对胃癌裸鼠模型的影响

Proteomic Study of Xiaotansanjie Recipe Intervened Human Gastric Tumor-bearing Mice

目的基于差异蛋白质组学探讨消痰散结方干预胃癌裸鼠模型的部分效应机制。方法20只BALB/c-nu/nu裸鼠随机分为模型组和干预组,每组10只,采用皮下移植人胃癌MKN-45瘤块制备胃癌裸鼠动物模型。建模成功后,模型组、干预组分别灌胃给予生理盐水、消痰散结方,剂量均为0.4mL/(只·d),14d后全部处死取材。双向凝胶电泳(2-DE)分离肿瘤组织总蛋白,基质辅助激光解吸电离时间飞行串联质谱及数据库查询鉴定差异表达蛋白质。结果获得分辨率高和重复性、稳定性好的肿瘤组织2-DE图谱。经质谱鉴定的30个差异表达蛋白质中与细胞周期、增殖、分化与凋亡作用相关的4个蛋白分别是钙调蛋白、钙周期素结合蛋白、热休克蛋白27、热休克蛋白60。与模型组比较,在干预组中热休克蛋白27表达明显增高,热休克蛋白60、钙调蛋白、钙周期素结合蛋白表达显著降低。结论对细胞周期、增殖、分化与凋亡相关蛋白的调节可能是消痰散结方治疗胃癌的效应机制。

Objective To observe and analyze the tumor tissue protein spectrum changes of MKN-45 gastric cancer cell bearing nude mice intervened by Xiaotansanjie Recipe on the platform of proteomics, and discuss its anti-gastric cancer mechanism. Methods Twenty BALB/c-nu/nu mice were randomly divided into model group and intervention group. MKN-45 gastric cancer cells were subcutaneously implanted into immune-incompetent nude mice in 2 groups. The model group and the intervention group was separately administrated with saline and Xiaotansanjie Recipe, which were delivered by intragastric administration with a dosage of 0.4 mL every mouse per day. All mice were sacrificed after 2 weeks. The total protein maps of the tumor tissue separated were observed and analyzed by two-dimensionl gel electrophoresis （2-DE）. The differentially expressed proteins were digested and identified by MALDI- TOF/TOF-MS mass spectrograph and database search. Results The 2-DE maps with good reproducibility and stability were obtained. Among the 30 differently expressed proteins, 4 were identified as calmodulin, calcyclln binding protein cycle, heat shock protein 27 and heat shock protein 60. They all related with cell proliferation, differentiation and apoptosis. Compared with the model group, heat shock protein 27 was obviously promoted, and calcyclin-binding protein, calmodulin, heat shock protein 60 were significantly depressed in the intervention group. Conclusion Xiaotansanjie Recipe maybe plays antitumor effects through regulating the proteins related with cell proliferation, differentiation and apoptosis.