Helicobacter pylori infection combined with DENA revealed altered expression of p53 and 14-3-3 isoforms in Gulo-/- mice
Helicobacter pylori infection combined with DENA revealed altered expression of p53 and 14-3-3 isoforms in Gulo-/- mice
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DOI：
10.1016/j.cbi.2013.09.002
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发表时间：
2013-11-25
影响因子：
5.1
通讯作者：
Kim, Gon Sup
中科院分区：
文献类型：
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作者：
Nagappan, Arulkumar;Park, Hyeon Soo;Kim, Gon Sup
与大多数其他哺乳动物不同，人体没有能力在体内合成维生素 C。因此，人类必须通过日常饮食来获取维生素C。 Gulo(-/-)小鼠品系已知存在维生素C缺乏症，其维生素C摄入量可以像人类一样通过饮食控制，这对于研究各种疾病的分子机制具有重要价值。在本研究中，我们建立了Gulo(-/-)小鼠模型，并采用免疫组化和蛋白质组学方法研究了幽门螺杆菌感染和DENA后Gulo(-/-)小鼠胃组织中差异表达的蛋白质。胃组织免疫组织化学分析结果显示，幽门螺杆菌感染和 DENA 治疗后，肿瘤抑制因子 p53 蛋白表达显着降低 (p < 0.05)，但信使 RNA (mRNA) 转录水平没有下降，14-3-3 epsilon、14-3-3 delta、Ki-67 和 cleaved caspase 3 表达显着增加 (p < 0.05)。 Gulo(-/-) 小鼠。此外，14-3-3异构体（14-3-3 epsilon、14-3-3 sigma、14-3-3 zeta和14-3-3 eta）的敲低显着增加了AGS细胞中的亚G1期（细胞凋亡的特征），并且还观察到细胞收缩、密度和核分裂等表型变化。蛋白质组分析显示，幽门螺杆菌感染后，14-3-3 sigma、14-3-3 eta 和原肌球蛋白 α-1 链下调，Hspd1 蛋白和 HSC70 上调，其次是 DENA。免疫组化和蛋白质组学分析结果表明，H. pylori改变了p53和14-3-3亚型的表达，DENA进一步增强了H. pylori的作用，这可能参与了Gulo(-/-)小鼠胃癌的发生和转移。 (C) 2013 作者。由爱思唯尔爱尔兰有限公司出版。保留所有权利。
Unlike most other mammals, human bodies do not have the ability to synthesize vitamin C inside of their own bodies. Therefore, humans must obtain vitamin C through daily diet. Gulo(-/-) mice strain is known with deficiency, in which vitamin C intake can be controlled by diet like human, and would be valuable for investigating the molecular mechanism of various diseases. In the present study, we established Gulo(-/-) mice model and investigated the differentially expressed proteins in stomach tissue of Gulo(-/-) mice after Helicobacter pylori-infected, and followed by DENA, using immunohistochemistry and proteomic approach. The results of immunohistochemistry analysis of stomach tissue showed that the tumor suppressor, p53 protein, expression was significantly decreased (p < 0.05) but not messenger RNA (mRNA) transcriptional level, and 14-3-3 epsilon, 14-3-3 delta, Ki-67 and cleaved caspase 3 expressions were significantly increased (p < 0.05) by H. Pylori infection, and followed by DENA treatment in Gulo(-/-) mice. Moreover, knockdown of 14-3-3 isoforms (14-3-3 epsilon, 14-3-3 sigma, 14-3-3 zeta; and 14-3-3 eta) were significantly increased sub-G1 phase (characteristics of apoptosis) in AGS cells and, phenotypic changes like cell shrinkage, density and cleaved nuclei were also observed. Proteome analyses showed that 14-3-3 sigma, 14-3-3 eta, and tropomyosin alpha-1 chain were down-regulated, and Hspd1 protein and HSC70 were up-regulated after H. Pylori-infection, and followed by DENA. The combined results of immunohistochemistry and proteomic analysis suggest that H. pylori altered the p53 and 14-3-3 isoforms expression and DENA further enhanced the H. pylori effect, which might be involved in carcinogenesis and metastasis of gastric cancer on Gulo(-/-) mice. (C) 2013 The Authors. Published by Elsevier Ireland Ltd. All rights reserved.