Annexin A3 may play an important role in ochratoxin-induced malignant transformation of human gastric epithelium cells
Annexin A3 may play an important role in ochratoxin-induced malignant transformation of human gastric epithelium cells
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膜联蛋白A3可能在赭曲霉毒素诱导的人胃上皮细胞恶性转化中发挥重要作用
DOI：
10.1016/j.toxlet.2019.07.002
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发表时间：
2019-10-01
影响因子：
3.5
通讯作者：
Cui, Jinfeng
中科院分区：
文献类型：
--
作者：
Wang, Juan;Jia, Xin;Cui, Jinfeng
赭曲霉毒素 A (OTA) 是最丰富的食品污染霉菌毒素之一，可能对人类致癌。我们之前证明，长期（40周）OTA暴露会在体外诱导人胃上皮细胞（GES-1）的恶性转化。然而，OTA诱发胃癌的具体机制很复杂。在本研究中，我们利用2-DE和基质辅助激光解吸电离飞行时间质谱（MALDI/TOF MS）结合生物信息学和免疫印迹，在体外研究GES-1和OTA恶性转化的GES-1细胞（OTA-GES-1T细胞）之间的差异表达蛋白。我们发现恶性转化后鉴定出四种差异表达蛋白，包括肌动蛋白、细胞质1（ACTB）、F-肌动蛋白加帽蛋白亚基α-1（CAPZA1）、膜联蛋白A3（ANXA3）、红细胞硫氧还蛋白过氧化物酶B（TPx-B）和纤维蛋白原βB（纤维蛋白原β）。在差异表达蛋白中，通过MTT实验、蛋白质印迹实验、细胞周期分析、伤口愈合实验、Transwell实验和OTA-GES-1(T)细胞集落形成实验分析Annexin A3的作用。结果表明，siRNA抑制Annexin A3可有效阻止OTA-GES-1(T)细胞的增殖、迁移和侵袭能力。总的来说，这项研究的结果将指导未来 OTA 致癌性的研究。
Ochratoxin A (OTA), one of the most abundant food-contaminating mycotoxins, is a possible carcinogen to humans. We previously demonstrated that long-term (40 weeks) OTA exposure induces the malignant transformation of human gastric epithelium cells (GES-1) in vitro. However, the specific mechanism underlying OTA-induced gastric carcinogenesis is complex. In the present study, we used 2-DE and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF MS) combined with bioinformatics and immunoblotting to investigate the differentially expressed proteins between GES-1 and OTA-malignant transformed GES-1 cells (OTA-GES-1T cells) in vitro. We found that four differentially expressed proteins were identified after malignant transformation, including actin, cytoplasmic 1 (ACTB), F-actin-capping protein subunit alpha-1 (CAPZA1), Annexin A3 (ANXA3), thioredoxin peroxidase B from red blood cells (TPx-B) and Fibrinogen beta B (Fibrinogen beta). Among the differentially expressed proteins, the effect of Annexin A3 was analyzed by MTT assay, western blot, cell cycle analysis, wound healing assay, Transwell assay, and colony formation assay in OTA-GES-1(T) cells. The results showed that inhibition of Annexin A3 by siRNA effectively prevented the proliferation, migration, and invasion abilities of OTA-GES-1(T) cells. Collectively, the results of this study will guide future research on OTA carcinogenicity.