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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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Background/Aim: 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 NFkB activation on Her-2 expression in MCF-7 breast adenocarcinoma cells. Methods: Stably transfected MCF-7 cell lines with pcDNA3.0, IkBoc T, c-FUP/control shRNA were established by FuGENE with the supplementation of G418 (500|-ig /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. Results: 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 kB (NFkB) activation. We demonstrated that the antiapoptotic transcription factor NFkB counteracted this cleavage through the induction of caspase-8 inhibitor, c-FLJR Conclusion: we propose a novel mechanism in which NFkB functions as a new antiapoptotic factor by counteracting TNF-a-triggered Her-2 cleavage.
机译：背景/目的：Her-2过表达与乳腺癌患者预后不良有关。现在，我们探讨了TNF-α治疗和/或NFkB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（500μg-ig/ ml）建立pcDNA3.0，IkBoc T，c-FUP /对照shRNA稳定转染的MCF-7细胞。 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：发现Her-2是caspase-8的新底物，肿瘤坏死因子α（TNF-α）刺激导致MCF-7乳腺癌腺癌细胞中caspase-8依赖的Her-2裂解而导致Caspase-8缺陷。核因子kB（NFkB）激活。我们证明了抗凋亡转录因子NFkB通过诱导caspase-8抑制剂c-FLJR来抵消这种裂解。结论：我们提出了一种新的机制，其中NFkB通过抵消TNF-a触发的Her-2裂解而作为一种新的抗凋亡因子。 。

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