IL-6 plays a critical role in the synergistic induction of human serum amyloid A (SAA) gene when stimulated with proinflammatory cytokines as analyzed with an SAA isoform real-time quantitative RT-PCR assay system
IL-6 plays a critical role in the synergistic induction of human serum amyloid A (SAA) gene when stimulated with proinflammatory cytokines as analyzed with an SAA isoform real-time quantitative RT-PCR assay system
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DOI：
10.1016/j.bbrc.2003.12.096
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发表时间：
2004-02-06
影响因子：
3.1
通讯作者：
Yoshizaki, K
中科院分区：
文献类型：
--
作者：
Hagihara, K;Nishikawa, T;Yoshizaki, K
血清淀粉样蛋白A（SAA）是继发性淀粉样变性中淀粉样蛋白A（AA）的前体蛋白，SAA 1主要参与AA淀粉样变性。我们建立了SAA同种型实时定量RT-PCR检测，发现β-2微球蛋白作为我们系统的内部对照比GAPDH和β-肌动蛋白更稳定。IL-6和IL-1 β或IL-6和TNF α，但不是IL-1 β和TNF α，诱导SAA 1和SAA 2基因的协同诱导。抗IL-6受体单克隆抗体可完全抑制IL-6、IL-1 β和TNF α三重刺激对SAA 1和SAA 2的协同诱导作用，而IL-1受体拮抗剂或抗TNF α单克隆抗体仅部分抑制HepG 2、Hep 3B和PLC/PRF/5细胞的协同诱导作用。虽然SAA 1启动子没有STAT 3共有序列，但JAK 2转录因子-AG 490使SAA 1基因表达降低至30%，表明STAT 3参与其中。我们能够证明，IL-6在人SAA基因的协同诱导中起着关键作用，当用促炎细胞因子刺激时。(C)2003年爱思唯尔公司All rights reserved.
Serum amyloid A (SAA) is known to be a precursor of amyloid A (AA) protein in AA (secondary) amyloidosis and SAA1 to be mainly involved in AA amyloidosis. We established an SAA isoform real-time quantitative RT-PCR assay and found that beta-2 microglobulin is more stable as an internal control than GAPDH and beta-actin for our system. Either IL-6 and IL-1beta or IL-6 and TNFalpha, but not IL-1beta and TNFalpha, induced the synergistic induction of SAA1 and SAA2 genes. Anti-IL-6 receptor monoclonal antibody completely inhibited the synergistic induction of SAA1 and SAA2 during triple stimulation with IL-6, IL-1beta, and TNFalpha, but, IL-1 receptor antagonist or anti-TNFalpha monoclonal antibody was only partially inhibited in HepG2, Hep3B, and PLC/PRF/5 cells. Although the SAA1 promoter has no STAT3 consensus sequence, the JAK2 inhibitor-AG490 reduced SAA1 gene expression to 30%, suggesting the involvement of STAT3. We were able to demonstrate that IL-6 plays a critical role in the synergistic induction of human SAA gene when stimulated with proinflammatory cytokines. (C) 2003 Elsevier Inc. All rights reserved.