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Identification of Key Proteins in Human Epithelial Cells Responding to Bystander Signals From Irradiated Trout Skin

机译:响应鳟鱼皮肤旁观者信号的人上皮细胞中关键蛋白的鉴定

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Radiation-induced bystander signaling has been found to occur in live rainbow trout fish ( Oncorhynchus mykiss ). This article reports identification of key proteomic changes in a bystander reporter cell line (HaCaT) grown in low-dose irradiated tissue-conditioned media (ITCM) from rainbow trout fish. In vitro explant cultures were generated from the skin of fish previously exposed to low doses (0.1 and 0.5 Gy) of X-ray radiation in vivo. The ITCM was harvested from all donor explant cultures and placed on recipient HaCaT cells to observe any change in protein expression caused by the bystander signals. Proteomic methods using 2-dimensional (2D) gel electrophoresis and mass spectroscopy were employed to screen for novel proteins expressed. The proteomic changes measured in HaCaT cells receiving the ITCM revealed that exposure to 0.5 Gy induced an upregulation of annexin A2 and cingulin and a downregulation of Rho-GDI2, F-actin-capping protein subunit beta, microtubule-associated protein RP/EB family member, and 14-3-3 proteins. The 0.1 Gy dose also induced a downregulation of Rho-GDI2, hMMS19, F-actin-capping protein subunit beta, and microtubule-associated protein RP/EB family member proteins. The proteins reported may influence apoptotic signaling, as the results were suggestive of an induction of cell communication, repair mechanisms, and dysregulation of growth signals.
机译:辐射诱导的旁观者信号已发现在活的虹鳟鱼(Oncorhynchus mykiss)中发生。本文报道了在虹鳟鱼低剂量照射组织条件培养基(ITCM)中生长的旁观者报告细胞系(HaCaT)中关键蛋白质组学变化的鉴定。从体外暴露于低剂量(0.1和0.5 Gy)X射线的鱼皮中产生体外外植体培养物。从所有供体外植体培养物中收获ITCM,并将其置于受体的HaCaT细胞上,以观察由旁观者信号引起的蛋白质表达的任何变化。使用二维(2D)凝胶电泳和质谱的蛋白质组学方法来筛选表达的新蛋白质。在接受ITCM的HaCaT细胞中测得的蛋白质组学变化表明,暴露于0.5 Gy会诱导膜联蛋白A2和环丁蛋白的上调,而Rho-GDI2,F-肌动蛋白封端的蛋白亚基β,微管相关蛋白RP / EB家族成员的下调和14-3-3蛋白。 0.1 Gy剂量还诱导了Rho-GDI2,hMMS19,F-肌动蛋白封端蛋白亚基β和微管相关蛋白RP / EB家族成员蛋白的下调。报告的蛋白质可能影响凋亡信号传导,因为结果提示诱导细胞通讯,修复机制和生长信号失调。

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