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首页> 外文期刊>Cellular Physiology and Biochemistry >Tumor Necrosis Factor α Stimulates Her-2 Cleavage by Activated Caspase-8
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Tumor Necrosis Factor α Stimulates Her-2 Cleavage by Activated Caspase-8

机译：肿瘤坏死因子α激活Caspase-8刺激Her-2切割。

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biBackground/Aim/i/b Her-2 over-expression has been correlated with a poor prognosis in patients with breast cancer. Now, we explored the effect of TNF-α treatment and/or NF?B activation on Her-2 expression in MCF-7 breast adenocarcinoma cells. biMethods/i/b Stably transfected MCF-7 cell lines with pcDNA3.0, I?Bα MT, c-FLIP/control shRNA were established by FuGENE with the supplementation of G418 (500 µg /ml). Western blot and Real-time PCR were applied to assess the expression levels of protein and mRNA of target gene. In addition, caspase-8 activity was evaluated by the incubation with a caspase-8 fluorogenic substrate, Ac-IEPD-AMC using a spectrofluorometer. biResults/i/b It was uncovered that Her-2 was a new substrate for caspase-8 and that tumor necrosis factor α (TNF-α) stimulation resulted in a caspase-8-dependent Her-2 cleavage in MCF-7 breast adenocarcinoma cells defective for nuclear factor ?B (NF?B) activation. We demonstrated that the antiapoptotic transcription factor NF?B counteracted this cleavage through the induction of caspase-8 inhibitor, c-FLIP. biConclusion/i/b we propose a novel mechanism in which NF?B functions as a new antiapoptotic factor by counteracting TNF-α-triggered Her-2 cleavage.
机译：背景/目标 Her-2过表达与乳腺癌患者的不良预后相关。现在，我们探讨了TNF-α治疗和/或NF？B激活对MCF-7乳腺癌细胞中Her-2表达的影响。 方法 通过FuGENE补充G418（500 µg），建立了以pcDNA3.0，I？BαMT，c-FLIP / control shRNA稳定转染的MCF-7细胞系。 / ml）。用Western blot和实时荧光定量PCR检测靶基因蛋白和mRNA的表达水平。另外，通过使用分光荧光计与caspase-8荧光底物Ac-IEPD-AMC一起温育来评估caspase-8活性。 结果 研究发现，Her-2是caspase-8的新底物，肿瘤坏死因子α（TNF-α）刺激导致caspase-8依赖性MCF-7乳腺腺癌细胞中的Her-2裂解对核因子？B（NF？B）激活有缺陷。我们证明了抗凋亡转录因子NF？B通过诱导caspase-8抑制剂c-FLIP抵消了这种切割。 结论 我们提出了一种新的机制，其中NF？B通过抵消TNF-α触发的Her-2裂解而作为一种新的抗凋亡因子。

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