Identification of copper/zinc superoxide dismutase as a nitric oxide-regulated gene in human (HaCaT) keratinocytes:: implications for keratinocyte proliferation
Identification of copper/zinc superoxide dismutase as a nitric oxide-regulated gene in human (HaCaT) keratinocytes:: implications for keratinocyte proliferation
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DOI：
10.1042/0264-6021:3460719
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发表时间：
2000-03-15
影响因子：
4.1
通讯作者：
Pfeilschifter, J
中科院分区：
文献类型：
--
作者：
Frank, S;Kämpfer, H;Pfeilschifter, J
最近的研究表明诱导型一氧化氮合酶表达的诱导与多种皮肤炎症性疾病相关。为了确定一氧化氮（NO）在皮肤中的作用机制，我们试图鉴定角质形成细胞中受一氧化氮调节的基因。使用人角质形成细胞系 HaCaT 作为模型系统，我们鉴定了一种 Cu/Zn 超氧化物歧化酶 (SOD)，该酶可被高浓度 (500 μM) 的 NO 捐赠剂（S-亚硝基谷胱甘肽、硝普钠和 (2)-1-[2(2-氨基乙基)-N-(2-氨基乙基)氨基]d 1-鎓-1,2-二醇(DETA-NO))，但不能通过血清或单一重组生长因子和炎性细胞因子或用超氧阴离子治疗。此外，内源产生的 NO 增加了角质形成细胞中 Cu/Zn SOD mRNA 的表达。此外，用NO处理HaCaT细胞与细胞增殖的双相效应相关，因为低剂量（100μM）的不同NO供体（S-亚硝基谷胱甘肽和DETA-NO）介导细胞增殖信号，而高浓度（500μM）则具有细胞抑制作用。为了确定角质形成细胞中 Cu/Zn SOD 表达与 NO 增殖之间可能的相关性，我们建立了一个细胞系 (psp1CZ1N)，该细胞系在松甾酮诱导型启动子构建体的控制下携带人 Cu/Zn SOD cDNA。 Ponasterone 诱导的 Cu/Zn SOD 过度表达对 psp1CZ1N 细胞增殖产生细胞抑制作用。因此，我们认为 NO 上调 Cu/Zn SOD 表达建立了角质形成细胞增殖的抑制机制。
Recent studies have demonstrated an induction of expression of inducible nitric oxide synthase that is associated with several inflammatory diseases of the skin. To define the mechanisms of action of nitric oxide (NO) in the skin, we attempted to identify genes that are regulated by NO in keratinocytes. Using the human keratinocyte cell line HaCaT as a model system, we identified a Cu/Zn superoxide dismutase (SOD) that was strongly induced by high concentrations (500 mu M) of NO-donating: agents (S-nitrosoglutathione, sodium nitroprusside and (2)-1-[2(2-aminoethyl)-N-(2-ammonioethyl)amino]d 1-ium-1,2-diolate (DETA-NO)), but not by serum or by single recombinant growth factors and inflammatory cytokines or by treatment with superoxide anions. Furthermore, endogenously produced NO increased the expression of Cu/Zn SOD mRNA in keratinocytes. Moreover, treatment of HaCaT cells with NO was associated with a biphasic effect on cell proliferation, because low doses (100 mu M) of different NO donors (S-nitrosoglutathione and DETA-NO) mediated a proliferative signal to the cells, whereas high concentrations (500 mu M) were cytostatic. To determine a possible correlation between the close regulation of Cu/Zn SOD expression and proliferation by NO in keratinocytes, we established a cell line (psp1CZ1N) carrying a human Cu/Zn SOD cDNA under the control of a ponasterone-inducible promoter construct. Ponasterone-induced overexpression of Cu/Zn SOD caused a cytostatic effect in proliferating psp1CZ1N cells. We therefore suggest that the up-regulation of Cu/Zn SOD expression by NO establishes an inhibitory mechanism on keratinocyte proliferation.