Regulation of the release of tumour necrosis factor (TNF)alpha and soluble TNF receptor by gamma irradiation and interferon gamma in Ewing's sarcoma peripheral primitive neuroectodermal tumour cells
Regulation of the release of tumour necrosis factor (TNF)alpha and soluble TNF receptor by gamma irradiation and interferon gamma in Ewing's sarcoma peripheral primitive neuroectodermal tumour cells
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DOI：
10.1007/s004320050055
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发表时间：
1997-05-01
影响因子：
3.6
通讯作者：
Jurgens, H
中科院分区：
文献类型：
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作者：
vanValen, F;KentrupLardong, V;Jurgens, H
本研究分析了尤文肉瘤(ES)/外周原始神经外胚层肿瘤(PPNET)12个细胞株在伽玛射线照射前后肿瘤坏死因子(TNF)α和可溶性肿瘤坏死因子受体(sTNF-R)的产生。对ES/pPNET细胞培养上清液进行肿瘤坏死因子α特异性扩增的酶联免疫吸附试验(EL ISA)、sTNF-R-p55和sTNF-R-p75的检测。肿瘤细胞系释放最少量的肿瘤坏死因子α，显著释放sTNF-R-p55(7/12个细胞系)，不释放sTNF-R-p75。伽玛射线照射可诱导(3/12)细胞分泌或上调(3/12)细胞分泌肿瘤坏死因子α，但不改变sTNF-R-p55和sTNF-R-p75水平。重组人干扰素-γ可显著增加辐射反应细胞株分泌的肿瘤坏死因子α，而对sTNF-R-p55和sTNF-R-p75水平无明显影响。重组人干扰素-γ影响辐射反应的大小，而不是敏感性。分泌的肿瘤坏死因子α具有生物活性，对WEHI-164细胞具有细胞毒作用，并可被抗肿瘤坏死因子α单抗中和。酪氨酸特异性蛋白激酶抑制剂赫比霉素A、蛋白激酶A抑制剂H89、磷脂酶A特异性抑制剂AA-COCF3和5-脂氧合酶特异性抑制剂MK-886不能抑制伽玛射线刺激的肿瘤坏死因子α的释放。抗氧化剂N-乙酰半胱氨酸、去甲二氢愈创木酸和甲平可剂量依赖性地抑制伽玛辐射介导的肿瘤坏死因子α的产生。综上所述，我们的研究结果表明，干扰素伽玛启动增强了ES/pPNET细胞对伽玛照射的反应，分泌具有生物活性的肿瘤坏死因子α，而不影响sTNF-R的释放。这些数据表明，在伽马辐射介导的细胞内信号通路中，需要蛋白酪氨酸激酶活性和活性氧物种，从而导致肿瘤坏死因子α的产生。
This study analyses the production of tumour necrosis factor (TNF)alpha and soluble TNF receptor (sTNF-R) before and after exposure to gamma irradiation and interferon gamma (IFN gamma) in 12 cell lines derived from Ewing's sarcoma (ES)/peripheral primitive neuroectodermal tumours (pPNET). Supernatants from ES/pPNET cell cultures were tested in a TNF alpha-specific amplified enzyme-linked immunosorbent assay (ELISA), a bioassay, and sTNF-R-p55 and sTNF-R-p75 ELISA. The tumour cell lines released minimal amounts of TNF alpha, prominent amounts of sTNF-R-p55 (7/12 cell lines) and no sTNF-R-p75. Exposure to gamma irradiation (5 Gy) either induced (3/12) cell lines) or up-regulated (3/12 cell lines) TNF alpha release without changing sTNF-R-p55 and sTNF-R-p75 levels. Priming of cultures with recombinant human IFN gamma (rhIFN gamma) markedly enhanced TNF alpha secretion in the radiation-responsive cell lines and had no influence on sTNF-R-p55 and sTNF-R-p75 levels. rhIFN gamma affected the magnitude rather than the sensitivity of the radiation response. The TNF alpha secreted was bioactive, as shown by its cytotoxic effect of WEHI-164 cells, and neutralization of its activity by anti-TNF alpha monoclonal antibody. Herbimycin A (a tyrosine-specific protein kinase inhibitor) but not calphostin C (a protein kinase C inhibitor), H89 (a protein kinase A inhibitor), AA-COCF3 (a specific inhibitor of phospholipase A(2)) and MK-886 (a specific inhibitor of 5-lipoxygenase) abrogated gamma-irradiation-stimulated TNF alpha release. The anti-oxidants N-acetylcysteine, nordihydroguaiaretic acid and mepacrine dose-dependently inhibited gamma-irradiation-mediated TNF alpha production. Collectively our findings indicate that IFN gamma priming potentiates the secretion of bioactive TNF alpha by ES/pPNET cells in response to gamma irradiation without affecting sTNF-R release. The data suggest a requirement for protein tyrosine kinase activity and a role for reactive oxygen species in the gamma-irradiation-mediated intracellular signalling pathway leading to TNF alpha production.